Yang Q Y, Li J H, Wang Q Y, Wu Y, Qin J L, Cheng J J, Qiu J
Department of Laboratory Medicine, Tenth People's Hospital of Tongji University, Shanghai, China.
Central Laboratory, Tenth People's Hospital of Tongji University, Shanghai, China.
Genet Mol Res. 2014 Dec 4;13(4):10269-78. doi: 10.4238/2014.December.4.21.
We examined whether metastasis-associated gene 1 (MTA1) promotes cell proliferation via DNA damage repair in ovarian cancer. MTA1 was successfully down-regulated using small interfering RNA in the epithelial ovarian cancer cell lines SKOV-3 and OVCAR-3. Cell growth was evaluated through MTT and colony formation assays. Fluorescence-activated cell sorting analysis was used to evaluate the distribution of cells in the cell cycle, and cytotoxicity assays were performed to study cell sensitivity to cisplatin. A neutral comet assay was used to measure levels of ionizing radiation-induced DNA damage in SKOV-3 cells, and Western blot analyses were carried out to examine the expression of key proteins involved in DNA damage repair pathways. MTA1 knockdown markedly inhibited cell growth and led to S phase cell cycle arrest. In addition, MTA1 depletion conferred sensitivity of ovarian cancer cells to cisplatin. Moreover, MTA1 depletion increased the level of ionizing radiation-induced DNA damage and caused irreparable damage, which was illustrated by a remarkable increase and persistent existence of a comet tail as well as protein expression levels of γH2AX, pRPA, and pChk1, all of which play critical roles in DNA repair. Thus, MTA1 promotes the proliferation of epithelial ovarian cancer cells by enhancing DNA repair.
我们研究了转移相关基因1(MTA1)是否通过DNA损伤修复促进卵巢癌细胞增殖。在卵巢上皮癌细胞系SKOV-3和OVCAR-3中,利用小干扰RNA成功下调了MTA1。通过MTT和集落形成试验评估细胞生长。采用荧光激活细胞分选分析评估细胞周期中细胞的分布,并进行细胞毒性试验以研究细胞对顺铂的敏感性。采用中性彗星试验测量SKOV-3细胞中电离辐射诱导的DNA损伤水平,并进行蛋白质印迹分析以检测DNA损伤修复途径中关键蛋白的表达。敲低MTA1显著抑制细胞生长并导致S期细胞周期阻滞。此外,MTA1缺失使卵巢癌细胞对顺铂敏感。而且,MTA1缺失增加了电离辐射诱导的DNA损伤水平并导致不可修复的损伤,这表现为彗星尾显著增加并持续存在,以及γH2AX、pRPA和pChk1的蛋白表达水平升高,这些蛋白在DNA修复中均起关键作用。因此,MTA1通过增强DNA修复促进卵巢上皮癌细胞增殖。