The apoptosis inducing effect of Glycosmis pentaphylla (Retz.) Correa and its influence on gene expression in hepatocellular carcinoma cell line, Hep3 B

Abstract

Ethnopharmacological relevance: Glycosmis pentaphylla (Retz.) Correa is used in Indian traditional medicine against jaundice and other liver disorders.

Aim of the study: The study aims to determine the in vitro anticancer and apoptosis inducing activity of Glycosmis pentaphylla in hepatocellular carcinoma cell line, Hep3 B.

Materials and methods: The cytotoxic and apoptosis inducing activity of the crude extract and active fractions were estimated on Hep3 B and RAW264.7 cell lines by MTT assay, Hoechst staining, DNA fragmentation, morphological studies, reverse transcription polymerase chain reaction and anti-poly-(ADP-ribose)-polymerase assays. The phytochemical profiling of active extract was done by TLC and HPTLC methods.

Results: Ethanol extract of Glycosmis pentaphylla was more effective than other extracts in reducing the proliferation of Hep3 B cells. As revealed by the results from DNA fragmentation, Hoechst staining, morphological studies, RT-PCR, PARP cleavage and gene expression studies, active extract induced apoptosis on Hep3 B cell line in concentration and time dependent manner with increase in the Bax/Bcl2 gene expression ratio. Chemo profiling data revealed the presence of flavonoid in the active fraction.

Conclusions: The study showed that major active component in the ethanol extract of Glycosmis pentaphylla is a flavonoid which induces apoptosis on cancer cell line, Hep3 B, by increasing the expression ratio of Bax/Bcl2 genes in a time and dose dependent manner.