Effect of RNA Interference of BID and BAX mRNAs on Apoptosis in Granulosa Cell-derived KGN Cells

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  • SAI Takafumi
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • MATSUDA Fuko
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • GOTO Yasufumi
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • MAEDA Akihisa
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • SUGIMOTO Miki
    Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan
  • GAO Hong-Mei
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • KABIR Abul Khair Mohammad Ahan
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • LI Jun-You
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan
  • MANABE Noboru
    Animal Resource Science Center, The University of Tokyo, Kasama 319-0206, Japan Research Center for Food Safety, The University of Tokyo, Tokyo 113-8657, Japan

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Abstract

In mitochondrion-dependent type II apoptosis, BH3-interacting domain death agonist (BID) and BCL-2-associated X protein (BAX) promote death ligand and receptor-mediated cell death. In porcine ovaries, the levels of BID and BAX increase in follicular granulosa cells during atresia. In the present study, to confirm the pro-apoptotic activity of BID and BAX in granulosa cells, we examined the effect of RNA interference of BID or BAX on apoptosis using a human ovarian granulosa tumor cell line, KGN. By reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, expression of BID and BAX was detected in KGN cells. Then, we suppressed BID and BAX mRNA expression in KGN cells using small interfering RNA (siRNA). When BID or BAX was suppressed, a significant decrease in the apoptotic cell rate was noted. In granulosa-derived cells, BID and BAX showed pro-apoptotic activity. These results suggest that BID and BAX act as signal-transducing factors in mitochondrion-dependent type II apoptosis.

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