RNAi knockdown of caspase-activated DNase inhibits rotenone-induced DNA fragmentation in HeLa cells

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• 作者：Tadamiki Tsuruta ; Kentaro Oh-hashi ; Yoshihito Ueno ; Yukio Kitade ; Kazutoshi Kiuchi ; Yoko Hirata
• 关键词：CAD ; ICAD ; Rotenone ; siRNA
• 刊名：Neurochemistry International
• 出版年：2007
• 出版时间：March 2007
• 年：2007
• 卷：50
• 期：4
• 页码：601-606
• 全文大小：425 K

文摘

Rotenone, an inhibitor of mitochondrial complex I, induces apoptosis in a variety of cells. However, little is known about endogenous endonucleases involved in rotenone-induced DNA fragmentation, a biochemical hallmark of apoptosis. We used a chemically modified siRNA which is thought to be more effective than a non-modified siRNA to study whether caspase-activated DNase (CAD) contributes to this phenomenon. Western blot analysis showed that CAD protein decreased to 8 % of control levels in human cervical carcinoma HeLa cells after a 48 h transfection of siRNA. Consistent with the reduction of the protein level, the siRNA was found to inhibit rotenone-induced DNA fragmentation. These results suggest that CAD is the endogenous endonuclease that mediates internucleosomal DNA degradation in rotenone-induced apoptosis.