Silencing of Ref-1 expression by retrovirus-mediated shRNA sensitizes HEK293 cells to hydrogen peroxide-induced apoptosis.
Redox factor-1 (Ref-1) is a multifunctional protein involved in DNA base excision repair (BER) and transcription factor modification. By the use of retrovirus-delivered shRNA, we effectively suppressed endogenous Ref-1 expression in human embryonic kidney (HEK) 293 cells. Our results showed that downregulation of Ref-1 rendered cells more sensitive to H(2)O(2)-induced apoptosis. In this process, the absence of Ref-1 decreased the ratio of Bcl-2/Bax protein expression, which resulted in cytochrome c release and caspase-3 activation. These data indicate that constitutive Ref-1 is required for cellular defense and that this protective function is dependent on its role in the regulation of Bcl-2 family proteins.
Xiong
Jian-Jun
JJ
Department of Biochemistry and Molecular Biology, Sun Yat-sen University, Guangzhou, China.
Zhang
Yan
Y
Wang
Jia-Li
JL
Bao
Guo-Dong
GD
Zhang
Yang
Y
Zhu
Zhen-Yu
ZY
D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D001483:Base Sequence; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D045304:Cytochromes c / Q000378:metabolism; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics* / Q000378:metabolism; D004789:Enzyme Activation; D020868:Gene Silencing; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity*; D018384:Oxidative Stress / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*; D051028:bcl-2-Associated X Protein / Q000235:genetics
D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D051028:bcl-2-Associated X Protein; D045304:Cytochromes c; D006861:Hydrogen Peroxide; D053148:Caspase 3; C085108:APEX1 protein, human; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase