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

• Published in: Journal of ethnopharmacology 2012-01, Vol.139 (2), p.359-365
• Main Authors: Sreejith, P.S., Mascarenhas, Russel Royna, Praseeja, R.J., Asha, V.V.
• Format: Article
• Language: English
• Subjects: Animals; Anticancer; Antineoplastic Agents, Phytogenic - chemistry; Antineoplastic Agents, Phytogenic - isolation & purification; Antineoplastic Agents, Phytogenic - pharmacology; Antineoplastic Agents, Phytogenic - toxicity; Apoptosis; Apoptosis - drug effects; Apoptosis - genetics; bcl-2-Associated X Protein - genetics; Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; Cell Survival - drug effects; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; Dose-Response Relationship, Drug; Flavonoid; Flavonoids - analysis; Gene Expression Regulation, Neoplastic - drug effects; Glycosmis pentaphylla (Retz.) Correa; Hep3 B; Humans; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Macrophages - drug effects; Mice; Plants, Medicinal; Poly(ADP-ribose) Polymerases - metabolism; Proto-Oncogene Proteins c-bcl-2 - genetics; Reverse Transcriptase Polymerase Chain Reaction; Rutaceae - chemistry; Time Factors
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2011.11.012

Glycosmis pentaphylla (Retz.) Correa is used in Indian traditional medicine against jaundice and other liver disorders. The study aims to determine the in vitro anticancer and apoptosis inducing activity of Glycosmis pentaphylla in hepatocellular carcinoma cell line, Hep3 B. 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. 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. 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.