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In vitro antibacterial evaluation of Moringa oleifera against wound infecting bacteria: Phytochemical based approach
Background: The rising trend of antibiotic resistance has made it essential to explore alternative antibacterial agents. Medicinal plants have been found to be a valuable source of natural antimicrobials, and this study sought to screen the antibacterial properties of methanolic extracts from leaves of medicinal plant Moringa oleifera against pathogenic bacteria Staphylococcus aureus (a Gram-positive bacterium) and Escherichia coli (a Gram-negative bacterium). Methods: Clinical wound samples containing Gram-positive and Gram-negative bacteria were obtained, sub-cultured and identified using Gram staining and standard biochemical tests. Methanolic extracts of M. oleifera leaves were prepared and screened for phytochemicals using standard qualitative assays. Antibacterial activity was evaluated by agar well diffusion and broth microdilution methods to determine zones of inhibition and minimum inhibitory concentrations. Data was analyzed using GraphPad Prism software. Results: Phytochemical analysis of M. oleifera methanolic leaf extract confirmed the presence of flavonoids, steroids and saponins, while quinones were absent. Bacterial isolates were identified as S. aureus (Gram-positive) and E. coli (Gram-negative) through morphological and biochemical tests. The extract exhibited concentration-dependent antibacterial activity, with negligible inhibition at lower concentrations and significant zones of inhibition at 1200 µL against both organisms. The minimum inhibitory concentration was determined to be as low as 300 µL, indicating effective antibacterial potential of the extract. Conclusion: This study proves the antibacterial properties of M. oleifera and indicates that the plant can serve as sources of natural antibacterial for treating bacterial infections.
Copyright (c) 2026 Authors publishing manuscripts with Biology Circle journals agree to transfer copyright to the journal and will grant journal’s right to publication under CC-BY 4.0 license. Authors are strongly encouraged to distribute the published work at their personal levels including social media pages, academic websites and institutional repositories etc. https://creativecommons.org/licenses/by/4.0
In vitro evidence supports cell cycle arrest by alkyl-phospholipid erufosine in pancreatic cancer cell line
Background: Pancreatic cancer is an aggressive malignancy with limited therapeutic options and poor prognosis. Dysregulation of cell cycle control is a hallmark of tumor progression, making cell cycle–targeted therapies an attractive strategy. Erufosine, a synthetic alkyl phosphocholine, has shown promising antitumor activity in various cancer models by interfering with signaling pathways involved in cell survival and proliferation. However, its effects on cell cycle regulation in pancreatic cancer cells require investigation. Methods: Human pancreatic cancer cells were treated with erufosine at defined concentrations for specified time interval. Cell cycle distribution was analyzed using propidium iodide-based staining of DNA followed by flow cytometry. The staining enabled quantification of DNA content to assess proportion of cells in different phases of the cell cycle. Results: Erufosine treatment resulted in alteration of cell cycle progression in pancreatic cancer cells. Flow cytometric analysis revealed an accumulation of cells in a specific phase of the cell cycle (G2/M), indicating a halt in cell cycle progression compared to untreated controls. This arrest was accompanied by a corresponding reduction in the proportion of cells undergoing active proliferation. Conclusion: These findings suggest that erufosine exerts its anti-tumor effects in pancreatic cancer, at least in part, by inducing cell cycle arrest. Erufosine may therefore represent a potential therapeutic agent targeting cell cycle dysregulation in pancreatic cancer.
Nanoparticles in Biology: Applications, Mechanisms and Future Directions
Nanoparticles have emerged as an evolutionary tool in the world of modern biology because of their extraordinary physiochemical characteristics, tunable surface chemistry, and ability to engage with biological systems at the molecular level. This review represents current innovations in the design, functionalization, and implementation of nanoparticles in an extensive range of biological fields. Their key applications include targeted drug delivery, bio-sensing, gene delivery, regenerative and personalized medicine, and remodeling of the cellular microenvironment. In this review, we will demonstrate basic interaction processes such as cellular uptake pathways, intracellular trafficking, nano-bio interface dynamics, and biodistribution, which play a vital role in the efficacy and safety of medicine. Particular attention is on biocompatibility, immunological mechanisms, and toxicity strategies in order to enhance selectivity and biological performance. Advanced directions, including smart stimuli-responsive nanoparticles, nano-bio hybrid systems, tailored bioinspired nanomaterials, and their uses in personalized and precise medicine, have been discussed. A detailed description of the present landscape and future applications of nanoparticles in biology, with an emphasis on their growing contributions in next-generation diagnostics and therapeutic advancements, has been demonstrated.
Renin-Angiotensin System Polymorphism in Chronic Kidney Disease and Diabetic Nephropathy
The renin-angiotensin system (RAS) is a vital hormonal pathway that regulates blood pressure, fluid balance and vascular tone but its dysregulation has been linked to several renal ailments including chronic kidney disease (CKD) and diabetic nephropathy (DN). Angiotensin II, the key effector molecule of RAS, acts through receptors AGTR1 and AGTR2 thus influencing cell proliferation, inflammation, and angiogenesis. Current review summarizes the key RAS gene variations associated with CKDs. Genetic polymorphisms in RAS components particularly ACE (I/D), AGTR1 (A1166C), and AGT (M235T) have been associated with increased susceptibility and progression of renal diseases. In CKDs, ACE DD and AGTR1 CC genotypes correlate with higher ACE activity, hypertension, and faster disease progression highlighting their diagnostic and therapeutic significance. Hyperglycemia induced RAS activation has been associated with DN through elevation of Ang II levels. The elevated Ang II leads towards oxidative stress and vascular injury. Overall, RAS genetic polymorphism plays a central role in renal disease pathogenesis, hence offering valuable insight for personalized medicine and targeted therapy.
Copyright (c) 2025 Authors publishing manuscripts with Biology Circle journals agree to transfer copyright to the journal and will grant journal’s right to publication under CC-BY 4.0 license. Authors are strongly encouraged to distribute the published work at their personal levels including social media pages, academic websites and institutional repositories etc. https://creativecommons.org/licenses/by/4.0
Serotype distribution and association of dengue viral load with clinical severity: A serotype-specific qRT-PCR study
Background: Dengue fever is a viral infection caused by arbovirus; an enveloped single standard RNA virus belongs to the family Flaviviridae. Since last two years, onward from 2023, changes in climate, urbanization and distribution pattern of biological vectors leads to increase transmission of dengue virus. In recent outbreak, all four serotype of dengue virus DENV-1, DENV-2, DENV-3 and DENV-4 have been detected in Pakistan. Methods: The current prospective cross-sectional study was conducted from March 2025 to October 2025 at Allama Iqbal Teaching Hospital Dera Ghazi khan. A total of 278 dengue suspect patients were enrolled in the study and were using serotype specific quantitative real-time polymerase chain reaction (qRT-PCR). Results: The mean age of the participant was 35 years ranging from 15 to 76 years and there was a male predominance (64.4%). The results of current study showed that among these 278 tested samples, 28 were positive for dengue virus as confirmed by qRT-PCR. Among the 28 confirmed dengue cases, DENV-2 was most prevalent serotype 16/28, followed by DENV-1 8/28, DENV-3 3/28 and DENV-4 was found to be least prevalent (1/28). Seasonal distribution analysis showed highest positivity in the months of August and September with 7 and 10 positive cases respectively. The results of present study also showed that the mean Ct value for DENV-1, DENV-2, DENV-3 and DENV-4 were 27.8 ± 4.6, 23.9 ±3.5, 28.1 ±5.0 and 28.4 ±4.4 respectively. It was observed that individuals infected with serotype DENV-2 had significantly lower Ct value which indicated higher viral load. Patients with higher viral load showed severe signs and symptoms as well. Conclusion: DENV-2 serotype was found to be the most prevalent serotype during 2025 dengue outbreak followed by DENV-1, DENV-3 and DENV-4 in studied region. Higher viral load was directly proportional to severity of disease particularly in patient infected with DENV-2.
Citrus limon based synthesis of zinc oxide nanoparticles: Antibacterial applications with improved outcomes
Background: Biosynthesis of nanoparticles by biological means, especially plant extract plays a major role in the field of nanotechnology. Zinc Oxide nanoparticles (ZnO-NPs) along with natural resources have demonstrated several advantages over traditional synthetic processes, including ease of use, low cost, eco-friendliness, nontoxic by products, and the absence of critical temperature and pressure conditions. Methods: In the present study, the aqueous extract of lemon (Citrus limon) peel, pulp and leaves was used for the green synthesis of ZnO-NPs using 0.1 M Zinc nitrate dihydrate (Zn (NO3) 2) as a precursor in alkaline condition using NaOH. UV-visible spectroscopy, Fourier Transform Infrared Spectroscopy (FTIR) analysis and Scanning Electron Microscopy (SEM) were used to characterize the structures. Disc diffusion method was used to evaluate the antibacterial effects of the synthesized nanoparticles. Results: ZnO-NPs obtained were characterized by UV-visible spectroscopy in which the surface plasmon resonance was 280 nm. FTIR analysis showed appropriate peaks for various chemical groups present in ZnO-NPs and stretching vibrations at 432 cm-1 indicating the presence of Zinc oxide bond in nanoparticles. SEM results showed spherical and well dispersed structures. Antibacterial property of ZnO-NPs, evaluated by the well diffusion assay, showed zone of inhibition in various pathogenic strains of Gram positive as well as Gram negative bacteria. It was observed that pulp-based nanoparticles had highest activity amongst all strains. Conclusion: Use of aqueous lemon extract as a capping agent would improve the antibacterial property of ZnO-NPs and can be developed as antibacterial agents against a wide range of microorganisms to control and prevent bacterial infections.
Biochemical and molecular evidence of β-lactamases in klebsiella species isolated from septic children admitted in a tertiary care hospital
Background: Klebsiella is a genus of Gram-negative rod-shaped bacteria with a prominent polysaccharide-based capsule. Antibiotic resistance patterns of Klebsiella species are often associated with molecular features. Purpose of this study was to detect Extended-Spectrum Beta-Lactamases (ESBL), AmpC Beta-Lactamases (AmpC) and Metallo-Beta-Lactamases (MBL) enzymes in Klebsiella species. Methods: Total 259 samples were collected from patients (≤15 years old) admitted in various wards and visiting outpatient department of The Children Hospital, Lahore. All samples were processed and screened for ESBL, MBL and AmpC enzymes. Molecular detection by PCR was done to detect their genotypes and results were analyzed using SPSS software version 25. Results: Out of 259 patients, ESBL positive, MBL positive and AmpC positive were 53 (20.5%), 64 (24.7%) and 86 (33.2%), respectively. The isolates found positive to screening test were further processed for molecular analyses. The results of the PCR amplification of the genes showed the presence of NDM-1 in 59 (22.8%), CIT in 30 (11.6%), SHV in 22 (8.5%), CTX-M in 14 (5.4%), EBC gene in 12 (4.6%) and TEM in 2 (0.8%) isolates. Two isolates co-produced all the three enzymes followed by 9 that co-produced MBL and ESBL, 4 isolates co-produced AmpC and MBL and 4 co-produced ESBL and AmpC. Conclusion: A large number of patients were found to have serious bacterial infections with significant number of ESBL, MBL and AmpC-production along with co-existence.
Prevalence and antibiotic resistance of Pseudomonas species isolated from ear specimens
Background: Ear infections are common medical conditions caused by various microbes including viruses, bacteria and fungi. Empirical antibiotic therapy is the preferred method for treatment of ear infection. Without knowing the etiology and antibiotic susceptibility pattern can increase antibiotic resistance. The aim was to find the prevalence of Pseudomonas species and its antibiotic susceptibility in ear specimens. Methods: A total of 726 patients with discharging ears attending ear, nose, throat (ENT) outpatient clinic from December 2019 to December 2020 were collected in Abbas Institute of Medical Sciences and associated clinical laboratories, Muzaffarabad. Pseudomonas species were isolated and characterized by biochemical and microbial techniques. Antibiotic susceptibility was estimated through Kirby -Bauer disc diffusion method. Results: Pseudomonas species were found in 13.3% (97/726) of the total samples tested. The prevalence of Pseudomonas was higher in adults (59/97=61%) as compared to children (38/97=39%). Moreover, the prevalence was higher in male (56/97=58%) than female (41/97=42%). Antibiotic sensitivity data revealed that Pseudomonas species were more sensitive towards Tazobactam (90.7%) followed by Imipenem (81.4%), Meropenem (76.2%), Amikacin (73.1%), Gentamycin (49.4%) and Tobramycin (37.1%). Sensitivity towards Ceftazidime, Cefepime, and Levofloxacin was found to be 31.9%, 24.7% and 21.6% respectively in isolates. Conclusion: Pseudomonas prevalence in ear infections is substantial in the region and should be considered in the patients having a history of acute and chronic discharge. Antibiotic susceptibility should be considered for such bacteria instead of use of empirical antibiotics to reduce resistance and treatment failure.
Antitumor potential of plant protein (Riproximin) against breast and colorectal cancer: Facts from functional and molecular investigations
Background: Ribosomal inactivating proteins comprise a big domain and inactivate the ribosomal machinery of the target cells irreversibly and induce toxic effects. Riproximin is a type II ribosomal inactivating protein which has been found lethal against various cancer cells originating from different tissues including breast and colorectal. The purpose of this study was to evaluate the effects of riproximin exposure on cell cycle relevant genes in breast and colorectal cancer cells. Methods: Toxic effects of riproximin in breast (MDA-MB-231, MCF-7) and colorectal cancer cell lines (SW480, SW620) were identified by MTT assay. AT molecular levels, the cell lines were treated with riproximin separately and expressional levels of the three S-phase cell cycle related genes (CDC6, MCM3, MCM5) were evaluated to identify the effects of riproximin. Untreated samples were used as controls throughout the study. Results: Riproximin induced toxic effects in breast and colorectal cancer cells (IC50<1.2ng/ml, 24 hours). Exposure of the cells with different concentrations of riproximin showed concentration dependent effects on expressional profile of the selected S-phase related genes. The selected genes were continuously down regulated by the protein in the breast and colorectal cancer cell lines. The riproximin associated effects were statistically significant. Conclusion: Riproximin imposed significant cytotoxic and gene modulation effects in breast and colorectal cancer cells. Further studies are required to understand the molecular features affected by riproximin in cancer cells.
Angiotensinogen gene (met235thr and thr174met), angiotensin-converting enzyme (intron 16 insertion/deletion) and angiotensin II type 1 receptor (A1166C) gene polymorphisms in patients with pre-eclampsia
Background: Pre-eclampsia (PE) is a specific disorder of pregnancy characterized by new onset of hypertension (systolic blood pressure ≥ 140 mm Hg or diastolic blood pressure ≥ 90 mm Hg) and proteinuria (˃0.3g /day), presenting after 20th weeks of gestation. PE affects 2 to 4% of all pregnancies worldwide. Methods: Ninety-two pre-eclamptic women and ninety-two controls were genotyped using PCR-RFLP and Allele specific PCR analysis. In the present study, we analyzed allelic and genotypic distribution of AGT gene (Met235Thr and Thr174Met), ACE I/D and AT2R1 (A1166C) polymorphisms in patients with PE and healthy controls. Genetic modelling and haplotype analysis was done for these SNPs. Results: Met235Thr (T704C) polymorphism under dominant and log additive models showed increased risk with pre-eclampsia and reached statistically significant levels for association with the PE but when it was adjusted after Bonferroni correction it became insignificant. While Thr174Met (C521T) polymorphism of AGT gene and AT2R1 A1166C polymorphism did not show any association with PE. ACE I/D polymorphism under recessive model was found significantly associated with the development of PE. While dominant and over dominant models showed protective effect which was also statistically significant. Conclusion: Met235Thr and ACE I/D polymorphisms are significantly associated with the development of pre-eclampsia, while AT2R1 A1166C and Thr174Met polymorphisms are not associated with the disorder in local population.
Primer Optimization for SNP Based PCR And Quantitative Real Time PCR Assays
Primer designing is critical for efficient polymerase chain reaction (PCR) and real time PCR, especially for quantification of real time PCR assays and single nucleotide polymorphism (SNP) for genotype analysis. Careful selection and optimization of primers is necessary for specific, reliable and efficient results in these methodologies. While detecting SNP, it is important for primers to amplify specifically the target region with minimal risk of non-specific amplification or primer dimer formation. In real-time PCR or quantitative PCR (qPCR), primer designing is more complex as primers must be compatible with both the target sequence as well as fluorescent reporter system used to monitor the amplification in real-time. Therefore, knowledge and understanding of primer designing and principles of optimization are necessary for researchers and analysts who are dependent on PCR-based methods for required results. This review provides a comprehensive overview of the key considerations to design primers in SNP-based PCR and real-time PCR assays, covering the important aspects such as primer length, melting temperature, GC content, and specificity. It also elaborates the use of specialized software tools for primer designing and the importance of experimental validation to ensure optimal assay performance.
Alkyl-phospholipid mediated toxicity and regulation of transcriptomic profile of cell cycle genes in liver cancer cells
Background: Hepatocellular carcinoma (HCC) is the primary malignancy of the liver that accounts for about 90% of liver cancers. HCC is the sixth most prevalent cancer globally and the third driving reason for cancer associated deaths. Most used therapeutic approaches for liver cancer include surgery, chemotherapy, immunotherapy and targeted therapy. For liver disease patients, new treatment compounds are needed for a better cure for this disease. The focus of proposed study was to evaluate cytotoxic and cytostatic potential of a 3rd generation alkyl-phospholipid (erufosine) against liver cancer cells. Methods: HepG2 liver cancer cells were exposed to different concentrations of erufosine (0.38-50µM) for 24-72 hours and effects on proliferation were determined by MTT dye reduction assay. Afterwards, the cells were exposed to three distinct concentrations of erufosine (5, 10, 25µM)) followed by total RNA extraction and cDNA synthesis. The synthesized cDNA samples were used to identify expressional alterations in our selected genes (CCNB2, CCND2, CDKN1A, and MCM4) by real-time PCR. Results: It was clearly noticed that a substantial growth inhibition was noticed in HepG2 cells after exposure to erufosine. The genes were altered in discrete fashion where cell cycle inhibitor (CDKN1A) was induced, while cell cycle promoters (CCNB2, CCND2 and MCM4) were down-regulated especially at high concentrations of compound. Conclusion: Erufosine showed substantial cytotoxic and anti-proliferative effects against the liver cancer cells. Expressional variations in multiple cell cycle related genes were imposed by erufosine.
Erufosine alters the genes associated with G2/M Phase of cell cycle in cancers: Molecular evidence from gene expression analysis
Background: Erufosine is a novel alkyl-phospholipid with promising anticancer activity. This study investigated the molecular mechanisms underlying the anticancer effects of erufosine on breast and colorectal cancer cells, with a focus on cell cycle regulation related genes. Methods: Breast (MDA-MB-231, MCF-7) and colorectal cancer (SW480, SW620) cells were treated with erufosine, and cell viability was assessed by MTT assay. Afterwards, the cell lines were exposed to IC25, IC50, and IC75 concentrations of the test compound for 48 hours. qRT-PCR was used to examine expression changes in G2/M phase-related genes including CCNA2, CCNB1 and CDKN3. Fold differences were calculated by using Livak method between the groups. Results: Erufosine exhibited potent cytotoxicity at micro-molar concentrations in the cell lines. In MDA-MB-231 cells, erufosine induced concentration-dependent downregulation of all three G2/M genes. MCF-7 cells showed minimal induction at low concentrations, followed by marked inhibition at higher concentrations. In colorectal cancer cell lines, erufosine consistently inhibited the three selected genes at all applied concentrations. Conclusions: Erufosine induces cytotoxic effects and alterations in G2/M phase related genes in breast and colorectal cancer cells. This study provides molecular evidence for the cell cycle related effects of erufosine and supports its potential as a novel therapeutic agent for cancers, particularly for metastatic phases of disease.
Copyright (c) 2024 Copyright (c) 2024 Authors publishing manuscripts with Biology Circle journals agree to transfer copyright to the journal and will grant journal’s right to publication under CC-BY 4.0 license. Authors are strongly encouraged to distribute the published work at their personal levels including social media pages, academic websites and institutional repositories etc. https://creativecommons.org/licenses/by/4.0
CXCR4 blockage induces cytotoxic effects in liver cancer cells
Background: Hepatocellular carcinoma (HCC) is a malignant tumor which arises from the liver cells (hepatocytes). Chronic inflammation followed by fibroses and cirrhosis ultimately leads to the development of liver carcinoma. Various treatment strategies for HCC include surgery, radiotherapy, targeted agents and chemotherapy. Despite the availability of diverse therapeutic options, 5-year survival rates are low (10-30%) especially in advanced stages of HCC. This, in turn, highlights the need to identify new therapeutic targets/compounds for HCC treatment. The purpose of this study was to evaluate the effects of blocking chemokine receptor (CXCR4) on cell proliferation and apoptosis related genes (FAS/FASL) in liver cancer cells (HepG2). Methods: CXCR4 receptors were blocked by using FDA approved respective antagonist (AMD3100) in HepG2 cells and effects on proliferation were identified. For this purpose, the cells were exposed to various concentrations (7.5-500µM) of the test compound and cell viability was monitored by MTT dye reduction assay for 24, 48 and 72 hours. Afterwards, HepG2 cells were exposed to three distinct concentrations of AMD3100 (50, 100, 200µM) in a separate experiment and expressional modulations in two apoptosis related genes (FAS and FASL) were identified by qRT-PCR methodology. Results: Blocking CXCR4 by using AMD3100 induced minimal growth inhibitory effects on the cells. Blockage of CXCR4 induced marginal up-regulation of FAS and FASL genes in the cells. Conclusion: Further investigations are needed to discover the effects of CXCR4 blockage on liver cancer cells.
Antineoplastic effects of erufosine in combination with oxaliplatin in colorectal cancer
Background: Combinations of the anticancer agents is a commonly practiced strategy in cancer treatment as it provides an efficient way to control the disease burden with reduced side effects. Finding novel compounds to be combined with the available chemotherapeutic drugs for better treatment of colorectal cancer is a much-needed research domain. In this context, alkyl-phospholipids comprise an attractive class of compounds which induce anticancer effects by targeting the cell surface membranes. Erufosine is a 3rd generation of alkyl-phospholipid class and interacts with the surface lipid molecules of the cell membranes to induce antineoplastic effects in cancer cells. Methods: Toxic effects of the selected alkyl-phospholipid (erufosine) and a commonly used chemotherapeutic agent (oxaliplatin) for colorectal cancer were identified in the three human cancer cell lines. For this purpose, the cells were exposed to various concentrations of the test compounds (single agent and combination) for 24-72 hours and cell viability was assessed. Afterwards, the cells were exposed to the distinct concentrations of the test compounds for 48 hours and expressional modulations in a cell cycle master regulator (P21) and stress marker (GADD45A) were identified by qRT-PCR methodology. Results: The results indicated that erufosine and oxaliplatin induced substantial anti-proliferative effects in colorectal cancer cells. The inhibitory effects of erufosine and oxaliplatin alone were significantly pronounced. In addition, synergistic anti-proliferative effects were observed when erufosine was combined with oxaliplatin. In addition, qRT-PCR data showed noteworthy potential of the test compounds to induce expression of P21 and GADD45-A genes at mRNA levels in the cells. Combination of erufosine and oxaliplatin induced P21 gene more prominently in the cells, especially in metastatic colorectal cancer cells. Conclusion: Erufosine and oxaliplatin impose substantial cytotoxic effects in the colorectal cancer cells. The combination of erufosine and oxaliplatin showed synergistic anti-proliferative effects. The compounds up regulated the expression of cell cycle inhibitor and cell stress marker (P21 and GADD45A) in colorectal cancer cells.
Cytotoxic effects and expressional modifications by sericin in liver cancer cells
Background: Liver cancer is the 3rd leading cause of cancer related deaths worldwide with increased risk in males than females. Many natural derivatives are being used as anticancer agents due to their antioxidant and cytotoxic potential. Silk sericin, extracted by the degumming of silkworm cocoon, has emerged as a novel potential anticancer compound. Due to increased elasticity, stability and ease of access made it an important protein for research in cancer studies. In this study, cytotoxic effects of sericin and its impact on expressional changes of selected genes in liver cancer cells were investigated. Methods: Cultured HepG2 cells were treated with extracted sericin by the degumming of local cocoons and commercially available purified sericin (0.03-1 mg/ml) for 24-72 hours. MTT dye reduction assay was used to determine the inhibitory effects of sericin on cultured HepG2 cells. Expressional analysis of GADD45A, GADD45B and CDKN1A was done by real-time PCR. Fold changes were determined by Livak method while comparing the results with untreated controls. Results: The cytotoxic effect of sericin (pure and extracted) was concentration dependent as seen by increasing concentration, the suppression of proliferation became more pronounced. The effects were more intense after the mid-interval exposure period (48 hour). GADD45A was inhibited moderately whereas GADD45B and CDKN1A were induced with increasing concentration of sericin. Conclusion: Proliferation of HepG2 cells was inhibited by sericin exposure. Sericin also showed potential to regulate the expression of genes involved in DNA repair, cell cycle progression, DNA damage and apoptosis.
Role of PTEN and LKB1 in cellular metabolism: Implications in cancer cell biology
Cellular metabolism is crucial for sustaining life, as it generates adenosine triphosphate (ATP) and provides essential building blocks for macromolecules. Recent research has expanded our understanding of metabolism, revealing its significant roles in cellular signaling networks related to immune responses, development, and aging. Cancer cells exhibit distinct metabolic characteristics due to reprogramming, particularly the Warburg effect, where they preferentially utilize glycolysis for energy production even in the presence of oxygen. This metabolic shift supports their uncontrolled proliferation and survival. This review explains the roles of two key tumor suppressor proteins, PTEN and LKB1, in regulating cellular metabolism and their implications for cancer biology. It aims to elucidate how these proteins influence metabolic pathways such as glycolysis and oxidative phosphorylation and to highlight their potential as therapeutic targets in cancer treatment. Dysregulation of LKB1 and PTEN due to mutations leads to enhanced glycolytic activity and metabolic reprogramming in cancer cells, contributing to tumorigenesis. LKB1 acts as a critical activator of AMP-activated protein kinase (AMPK), promoting energy homeostasis and regulating cellular metabolism under stress conditions. Conversely, PTEN functions as a negative regulator of the PI3K/AKT/mTOR signaling pathway, preventing uncontrolled cell growth. Understanding the mechanisms by which LKB1 and PTEN regulate cellular metabolism is essential for developing targeted anticancer therapies that exploit the unique metabolic vulnerabilities of cancer cells. By focusing on these pathways, researchers can identify new strategies to improve treatment efficacy while minimizing harm to normal tissues.
Inhibition of pro-platelet basic protein (CXCL7) induces antineoplastic effects in colorectal cancer cells
Background: Colorectal cancer (CRC), a 3rd leading cause of cancer-related deaths worldwide, is characterized by high metastatic potential and poor survival rates in advanced stages. Pro-platelet basic protein (CXCL7) is a chemokine and plays crucial roles in promoting tumor progression, angiogenesis, immune modulation and metastasis. Elevated CXCL7 expression correlates with increased tumor vascularization, immune cell recruitment, and poor clinical outcomes. This study investigates the potential of targeting CXCL7 to inhibit CRC progression and metastasis related properties. Methods: The role of CXCL7 in CRC was evaluated by using SW480 (human) and CC531 (rat) cell lines. CXCL7 expression was knocked down using siRNA, and its effects on cell proliferation, colony formation, migration, wound healing and apoptosis were assessed through MTT assays, colony formation assays, migration assays, scratch-healing assays and nuclear staining, respectively. RNA expression levels were analyzed through real-time PCR. Results: CXCL7 knockdown was achieved significantly at mRNA levels, with a maximum reduction of 76% and 81% in SW480 and CC531 cells, respectively, within 24 hours. This led to reduced cell proliferation, where SW480 cells showed a maximum inhibition of 56% compared to 39% in CC531 cells. Colony formation declined significantly in both cell lines, with a more pronounced effect on large colonies. Migration and scratch healing capacities were reduced by up to 26% in SW480 cells and 67% in CC531 cells, demonstrating impaired motility. Apoptotic markers, including nuclear condensation and fragmentation, were observed after CXCL7 inhibition. Conclusion: Targeting CXCL7 in CRC reduces tumor cell proliferation, colony formation, migration, and wound healing while inducing apoptosis. These findings highlight CXCL7 as a promising therapeutic target for CRC management and warrant further exploration in clinical settings.
Effects of a ribosome inactivating plant protein (riproximin) on transcriptomic profile of apoptosis pathway genes in breast cancer cells
Background: Treatment options for breast cancer are limited and the available possibilities are largely palliative in nature. In this context, searching for more effective therapeutic compounds is inevitable. Riproximin, a plant derived ribosomal inactivating protein, is one such kind of anticancer compound and has shown substantial anticancer effects while altering the major components of different molecular pathways including cell cycle, autophagy, cell stress and apoptosis. The purpose of this study was to investigate the effects of riproximin exposure on expressional levels of genes associated with apoptotic pathway in breast cancer cell lines. Methods: In first phase, breast cancer cell lines (MDA-MB-231 and MCF-7) were exposed to riproximin and toxic effects were identified by MTT dye reduction assay. Afterwards, the cell lines were exposed to riproximin in a separate experiment and expressional modulations of multiple genes were evaluated via real-time PCRs. Fold changes in this study were calculated by Livak method (2∆∆Ct) while using untreated cells as controls. Results: Proliferation (MTT) assay showed that riproximin induced promising cytotoxic effects in breast cancer cell lines. MCF-7 cells were more responsive towards riproximin exposure as compared to MDA-MB-231 cells. Expressional assessment of the selected genes confirmed the potential of riproximin to alter the levels in breast cancer cells. Specifically, in response to riproximin exposure, three most effectively induced genes in MDA-MB-231 cells were BIK (83fold), TNFSF9 (59fold) and NFKB1 (20fold). In MCF-7 cells, three most up-regulated genes after riproximin treatment were TNF (129fold), LTA (58fold) and TNFSF9 (17fold). Conclusion: Riproximin bears significant cytotoxic potential against breast cancer cell lines. Expressional modulations in multiple apoptotic related genes are imposed by riproximin in the breast cancer cells. Further in vitro and in vivo studies are required to support the evaluation of riproximin in clinical settings against breast cancer cells, while targeting the apoptotic routes for cancer treatments.
Copyright (c) 2024 Copyright (c) 2024 Authors publishing manuscripts with Biology Circle journals should agree to transfer copyright to the journal and will grant journal’s right to publication under CC-BY 4.0 license. Authors are strongly encouraged to distribute the published work at their personal levels including social media pages, academic websites and institutional repositories etc. https://creativecommons.org/licenses/by/4.0
Neureglin 3 gene intronic polymorphisms rs10883866, rs6584400, rs1937970 and rs677221 in patients with Schizophrenia
Background: Schizophrenia is a progressive disease presenting with insufficiencies in thought processes, perceptions, and emotional responsiveness with early onset. There are no laboratory tests or specific biomarkers established so far to diagnose the disease, so clinical and psychiatric analysis are used worldwide. Abnormal neurodevelopment, genetic predisposition and environmental factors are the main factors associated with this disease. The Neuregulin 3 (NRG3) gene encodes the Neuregulin 3 protein which binds to the extracellular domain of ERBB4 receptor tyrosine kinase and controls the proliferation, differentiation, and migration of neural progenitor cells. Methods: Purpose of this study was to find the NRG3 gene polymorphisms i.e., rs10883866, rs6584400, rs1937970 and rs677221 in Schizophrenic patients (n=114) and then compared with healthy controls (n=114). For this purpose, blood samples were taken after the informed consent of patients and the controls followed by DNA extraction and genotyping through PCR-RFLP and PCR-ARMS. Results: The minor allele frequencies of the SNPs rs10883866, rs6584400, rs1937970 and rs677221 in schizophrenic patients were 21.1%, 28.1%, 68.1% and 62.2% respectively. The SNPs rs10883866 and rs6584400 in protective effect toward the Schizophrenia whereas, the other two SNPs showed any statistically significant difference in the allelic distribution between cases and controls. Haplotype block analysis revealed that C-G haplotype in block 1, A-G in block 2 had frequencies of 62.4% and 60.7% in cases. The haplotype G-A in block 1 showed the protective effect towards the disease Schizophrenia with the p-value 0.011. Conclusion: NRG3 has association with the disease schizophrenia. More studies should be conducted including different phenotypic traits of schizophrenia and meticulous clinical evaluation of the patients with larger sample size is suggested to find out the association of NRG3 gene with the schizophrenia.
Cytotoxic effects of alkyl-phospholipids (Erufosine and Perifosine) in combination with cell cycle check point inhibitor (ATM) in breast cancer cells
Background: Breast cancer is the most common cancer in females with continuously growing incidence over the last two decades. In advanced stages, the disease is difficult to manage and imposes a major morbidity and mortality burden. Available treatment options are limited with a moderate capacity to cure. Situation demands to look for alternative therapeutic compounds. Alkyl-phospholipids (ALPs) are attractive options, and the two latest generations of this class (erufosine and perifosine) have shown substantial anticancer potential against cancer. In addition, ATM, an important factor during DNA damage/response, has come up for its corresponding therapeutic relevancy. Combining the two above-mentioned classes can lead to effective therapeutic options against breast cancer. Methods: Toxic effects of the selected compounds on breast cancer cell lines were determined by MTT dye reduction assay. The selected cell lines (MDA-MB-231 and MCF-7) were cultured in 96-well plates and exposed to various concentrations of the compounds for three different time intervals (24, 48 and 72 hours). Following the exposure of cells with the agents, expression modulations in CDKN family of genes were monitored at transcriptome level via real-time PCR methodology. Results: Exposure with the ALPs and ATM inhibited cell proliferation of the two breast cancer cell lines. Overall, ALPs were more capable of inhibiting cell proliferation. Combination of ALPs and ATM worked synergistically and reduced the cellular proliferation more effectively especially in MCF-7 cells. ALPs and the selected ATM inhibitor induced the expression of CDKN family members. Overall, the induction was maximally seen in CDKN1A gene with a maximum induction level of 40fold in MCF-7 cells. Conclusion: ALPs and ATM have the potential to inhibit proliferation of breast cancer cells and can support each other synergistically. Important cell cycle inhibitor genes like CDKN family can be induced by using above mentioned compounds.
Chemokine expression modulations in primary and metastatic colorectal cancer
Background: Chemokine are small proteins primarily involved in cell movement, immune response, inflammation and tissue repair. Chemokine also plays an important role in disease conditions including cancer growth. The objective of this study was to analyze expressional modifications in the chemokine in primary and metastatic colorectal cancer by using clinical isolates and in vivo model respectively. Methods: Serum samples from a total of 40 colorectal cancer patients (stage II and III) and healthy controls were collected following the due ethical codes. Circulatory levels of the chemokine (CCL family) were quantified using enzyme-linked immunosorbent assay (ELISA). To investigate metastasis associated expression changes of the chemokine, colorectal cancer animal model and microarray methodologies were exploited. Results: Increased circulatory levels of CCL2, CCL17 and CCL22 were detected in stage II and III colorectal cancer patients when compared with healthy controls. In contrast, CCL11 levels were substantially low in colorectal cancer patients. As far as rat animal model for colorectal cancer liver metastasis is concerned, only CCL2 levels were considerably high during the early phase of colorectal cancer cells’ implantation (Day 03) into the liver. Other chemokines did not demonstrate noticeable de-regulation during the whole period of experimental procedures (up to Day 21). Conclusion: Differential circulatory levels of chemokine were detected in colorectal cancer patients and healthy controls. Liver metastasis of colorectal cancer cells was accompanied by a marked increase of CCL2 during early phase of settlement. The findings reflected a discrete change in the circulatory and metastasis associated with chemokine profile in colorectal cancer.
Effects of synthetic alkyl-phospholipid (Perifosine) on survival and cell cycle relevant genes of hepatocellular carcinoma cells
Background: Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality globally, particularly in Asia. Limited treatment options highlight the needs for novel therapies. Alkyl-phospholipids (ALPs) offer a promising alternative to traditional chemotherapy by targeting cell membrane integrity and signaling pathways rather than DNA replication. Perifosine, a synthetic ALP, has shown cytotoxic effects in cancer cells, but its mechanisms on cell cycle regulation in HCC require further investigation. Methods: This in vitro study evaluated the effects of perifosine on HepG2 liver cancer cells. Cytotoxicity of perifosine was assessed using the MTT assay while applying various concentrations over 24-, 48-, and 72-hours intervals. Gene expression analysis of cell cycle regulator genes (CCNA1, CCNA2, and CCND1) was conducted using qRT-PCR after treating the HepG2 cancer cells with perifosine at IC25, IC50, and IC75 concentrations for 48 hours. Results: Perifosine demonstrated time and dose dependent cytotoxicity, with increased cell death at higher concentrations and prolonged exposure. Perifosine exposure imposed discrete expression modifications in CCNA1, CCNA2 and CCNDI genes. Maximum de-regulation was observed for CNNA1 (8fold) and CCNA2 (-3.5fold) in response to perifosine exposure. Conclusion: Perifosine exhibited cytotoxic effects and imposed de-regulation in cell cycle related genes in liver cancer cells.
Cancer stem cells: Review of current state and future directions
Cancer is a group of disorders characterized by the abnormal proliferation of cells. Cancer can infiltrate or spread to different areas of the body. Cancer stem cells (CSCs) are small groups of cancer cells that have the ability to self-renew and develop into tumors by different signaling pathways. CSCs play a significant role in the development of many types of cancer and are a significant factor in tumor metastasis and relapses. They are usually resistant to the existing treatments primarily focusing on most tumor cells. Recent technological advances have made it easier to study the basic role of CSCs in cancer biology and target them for potential therapeutic strategies. In this minireview, we give a brief overview of recent insights into the characteristics, mechanisms, current treatment opportunities, challenges, and future directions of cancer stem cells and their potential to revolutionize cancer treatment.
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