Alkyl-phospholipid mediated toxicity and regulation of transcriptomic profile of cell cycle genes in liver cancer cells

Abstract

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 3^rd 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.