Mechanism Study on Induction of Apoptosis by Diosgenin-3-O-α-L-Rhamnopyranosyl-(1→4)-β-D-Glucopyranoside (DRG) in HCT-15 Cell in Vitro

WNG Sn-long;LUO Hong-mei;CI ing;CUI Cheng-in;LIU Hong-wei;WU Chun-fu

Chinese Pharmaceutical Journal ›› 2011, Vol. 46 ›› Issue (15) : 1167-1172.

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Chinese Pharmaceutical Journal ›› 2011, Vol. 46 ›› Issue (15) : 1167-1172.

Mechanism Study on Induction of Apoptosis by Diosgenin-3-O-α-L-Rhamnopyranosyl-(1→4)-β-D-Glucopyranoside (DRG) in HCT-15 Cell in Vitro

  • WANG San-long1，LUO Hong-mei2，CAI Bing3，CUI Cheng-bin4， LIU Hong-wei5， WU Chun-fu5
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Abstract

OBJECTIVE To investigate the molecular mechanism of the induction of apoptosis by Chinese medicine monomer diosgenin-3-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranoside (DRG) in HCT-15 cell in vitro. METHODS The apoptotic mechanism induced by DRG was investigated by Western blotting， in vitro Bcl-xL competitive binding assay and reverse transcription-PCR (RT-PCR) assay. RESULTS It was proved that DRG triggered a mitochondria-controlled apoptotic pathway to induce apoptosis in HCT-15 cells， which involved the release of cytochrome C from mitochondria into the cytosol in a time-dependent manner， the up-regulation of the ratio of Bax/Bcl-2 expression， and the cleavage of caspase-9， caspase-3 and PARP， without effect on the expression of P53， Bcl-xL and the cleavage of caspase-7. Whereas， the changes of proteins related to death receptor pathway such as FADD and caspase-8 were not detected. In addition， DRG was not found to inhibit the BH3 peptide from combining with Bcl-xL protein. Semi-quantitative RT-PCR indicated the changes of mRNA levels of Bcl-2 and Bax genes， which might contribute to the decreased protein levels of Bcl-2 and the increased protein levels of Bax caused by DRG， revealing that both Bcl-2 and Bax gene were involved in the apoptosis induced by DRG. CONCLUSION The molecular mechanism of DRG inducing HCT-15 cells apoptosis involves in such processes as affecting the mRNA transcription level of Bax and Bcl-2 genes， up-regulating the ratio of Bax/Bcl-2 protein expression level and at last activating classical mitochondrial pathway but not death receptor pathway and P53-independent manner to induce cell apoptosis.

Key words

apoptosis / HCT-15 cell / Dioscorea futschauensis R. Kunth / diocgenin-3-α-L-rhamnopyranosyl-β-D-glucopyranoside (DRG) / mitochondrial pathway / death receptor pathway

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WNG Sn-long;LUO Hong-mei;CI ing;CUI Cheng-in;LIU Hong-wei;WU Chun-fu. Mechanism Study on Induction of Apoptosis by Diosgenin-3-O-α-L-Rhamnopyranosyl-(1→4)-β-D-Glucopyranoside (DRG) in HCT-15 Cell in Vitro[J]. Chinese Pharmaceutical Journal, 2011, 46(15): 1167-1172

References


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