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Gecko Skin Extract Induce Cytotoxicity and Apoptosis in Human Breast Cancer Cell Line

Tanmoy Bhowmik, Dilip Muhuri, Ajay Kumar Biswas, Aparna Gomes and Antony Gomes*

Gecko has been used against malignant tumors in medicinal practice in china, especially digestive system tumors such as esophageal cancer, gastric cancer, and liver cancer. Reduce inflammation, active against allergic responses, swelling having low toxicity are other activity of Gecko skin extract, has been mentioned in chinese folk medicine. The aim of this study was to examine the alcoholic skin extract of Gecko species induced cytotoxicity and apoptosis activity in breast cancer (MCF7) cell line. in vitro, the inhibitory effect of gecko skin alcohol extract (GSAE) on cytotoxicity of human breast cancer cells was measured by MTT assay and trypan blue exclusion assay which resulted in MCF7 cell proliferation was significantly inhibited in both dose- and time- dependent manner (P < 0.01). High percentage (71.19%) early cell apoptosis in GSAE-treated group (IC50 dose) exhibited selective cytotoxic action of GSAE confirmed by PE-Annexin V/FITC assay. Flow cytometry study showed that MCF7 cell cycle was arrested at G1 phase. Immunocytochemical study revealed that GSAE down regulated nuclear proliferative antigen Ki-67. Gecko alcohol extract decreased the potential of mitochondrial membrane and increased the expression of Bax, cytochrome c, caspase9, caspase3 and Ca2+ in cytosol. GSAE also downregulated the expression of Bcl2, HSP70 and HSP90. Although the level of VEGF did not change significantly, the expression of MMP9 was remarkably down-regulated in GSAE treated group. Thus GSAE showed significantly inhibitory effect in MCF7 cell line and acts through mitochondrial intrinsic pathway of cell apoptosis with downregulation of nuclear proliferative protein and heat shock protein.