Praseeja R J, PhD Student
Screening of anti-HCC activity of Cuscuta reflexa and its chemo sensitising effects
Hepatocellular carcinoma (HCC) is a type of primary liver cancer associated with high mortality rate. Development of an effective therapeutic agent is needed to prolong the life of HCC patients. Cuscuta reflexa Roxb.is used in the folk medicine to treat various disorders including weakness of liver, jaundice etc. We have examined the anti-HCC effect of C. reflexa on the HCC cell line, Hep 3B. The cytotoxic effect of C. reflexa extract on Hep 3B and HEK-293 cell lines were checked using MTT assay. The apoptosis inducing ability was studied using Hoechst staining, DNA fragmentation assay and Annexin V-FITC apoptotic assay. The molecular mechanism behind apoptotic induction was analysed using semi quantitative RT–PCR and Western blotting. Cell cycle analysis was done to find the effect of the extract on cell cycle progression. Finally a preliminary phytochemical investigation using TLC and HPTLC were also done. Treatment with this extract significantly increased the percentage of apoptotic cells. It also caused G1 arrest, which may be associated with apoptotic induction. The active fractions present in CRCE were also isolated. Our studies showed that the active extract of C. reflexa could reduce the proliferation and induce apoptotic cell death in Hep 3B cells. Thus, this is a promising plant for further studies towards anti-HCC drug development. For this, further studies on the isolation and characterisation of active principle need to be done.