Liu Guo-yan, Qu Quan-xin, Mi Ruo-ran, Qi Jing
Department of Obstetrics and Gynecology, General Hospital of Tianjin Medical University, Tianjin 300052, China.
Zhonghua Fu Chan Ke Za Zhi. 2006 May;41(5):339-42.
To study changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing (ERCC) 1 gene in ovarian cancer cell lines.
The small interference RNA (siRNA) targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to ovarian cancer cell line ES-2. The mRNA and protein of ERCC1 were evaluated by means of RT-PCR, western blot and immunocytochemistry. The changes of cisplatin sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTT) assay.
In ES-2 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfection. The sensitivity to cisplatin of ES-2 cell line was increased by 53.88 times after disturbing the ERCC1 gene.
The sensitivity to cisplatin of ovarian cancer cell lines ES-2 could be enhanced by RNA interfering ERCC1 gene.
通过RNA干扰卵巢癌细胞系中的切除修复交叉互补（ERCC）1基因，研究顺铂敏感性的变化。
设计并体外转录合成靶向ERCC1基因的小干扰RNA（siRNA），转染至卵巢癌细胞系ES-2。采用逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法（western blot）及免疫细胞化学法检测ERCC1的mRNA和蛋白。采用甲基噻唑基四氮唑（MTT）法检测干扰后顺铂敏感性的变化。
在ES-2细胞中，转染后24、48和72小时，ERCC1的mRNA和蛋白水平显著降低。干扰ERCC1基因后，ES-2细胞系对顺铂的敏感性提高了53.88倍。
RNA干扰ERCC1基因可增强卵巢癌细胞系ES-2对顺铂的敏感性。
