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KMID : 0644120040170020011
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Journal of Oriental Gynecology
2004 Volume.17 No. 2 p.11 ~ p.26
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Hangreunghwan water extract induced apoptosis in Human Cervical carcinoma HeLa cells.
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Yoo Sim-Keun
Kim Song-Baeg
Cho Han-Baek
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Abstract
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To address the ability of Hangreunghwan(HRH) to induce cell death, we investigated the effect of HRH on cell viability. Twenty-four hours later, loss of viability occurred following HRH exposure in a dose-dependent manner. The treatment of HRH, a commonly used herb formulation in Korea, Japan and China, caused a decrease in cell viability. HRH also resulted in apoptotic morphology a brightly blue-fluorescent condensed nuclei by Hoechst 33258-staining, and reduction of cell volume. Our results show that 2mg/ml HRH induces mitochondria membrane potential collapse. Immunoblotting data also shows that the expression of Bcl-2, antiapoptotic protein, decrease by the addition of HRH. This GFP-Bax overexpression system shows that an important pro-apoptotic Bcl-2-family protein, Bax is translocated to mitochondria by the addition of 2 mg/ml HRH. Interestingly, the treatment of BAPTA-AM, calcium chelator, shows that calcium is involved in HRH-induced apoptosis including caspase-3 activation in HeLa cells. BAPTA-AM reverses HRH-induced inhibition of XIAP expression. In addition, the calcium chelator inhibits the increase of Bax expression which is induced by HRH. However, the treatment of BAPTA-AM does not have any effect on HRH-induced Bax translocation. In conclusion, calcium is involved in both HRH-increased Bax expression and HRH-regulated XIAP expression, resulting in apoptosis in HeLa human cervical carcinoma cells.
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KEYWORD
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Hang-Reung-Hwan (HRH), Bax, mitochondria membrane potential, mitochondria, apoptosis, HeLa cells
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