Gao Yun, Su Dan, Ying Lisha, Lv Wangxia, Ma Shenglin
Cancer Research Institute, Zhejiang Cancer Hospital, Hangzhou 310022, China.
Zhongguo Fei Ai Za Zhi. 2010 Sep;13(9):846-9. doi: 10.3779/j.issn.1009-3419.2010.09.02.
The excision repair cross-complementing gene 1 (ERCC1), which is important in the repair of cisplatin-DNA adducts, was reported to be related to cisplatin resistance in tumor cells. The aim of this study is to investigate the changes of cisplatin sensitivity by silencing ERCC1 gene in lung cancer cell.
The small interfering RNA (siRNA) targeting ERCC1 gene was designed and synthesized, and transfected to lung cancer cell A549/DDP. The mRNA and protein expression levels of ERCC1 were evaluated by RT-PCR and Western blot. The changes of cisplatin sensitivity after RNA interference were examined by methyl thiazolyl assay.
In A549/DDP cell, the mRNA and protein levels of ERCC1 were decreased and the sensitivity to cisplatin was increased from 12.49 μg/mL to 9.27 μg/mL after transfection.
The sensitivity to cisplatin of lung cancer cell A549/DDP could be enhanced by RNA interfering ERCC1 gene targeted code 346.
切除修复交叉互补基因1（ERCC1）在顺铂-DNA加合物的修复中起重要作用，据报道其与肿瘤细胞对顺铂的耐药性有关。本研究旨在探讨通过沉默肺癌细胞中的ERCC1基因来改变顺铂敏感性。
设计并合成靶向ERCC1基因的小干扰RNA（siRNA），并转染至肺癌细胞A549/DDP。通过RT-PCR和蛋白质免疫印迹法评估ERCC1的mRNA和蛋白质表达水平。采用甲基噻唑基四唑法检测RNA干扰后顺铂敏感性的变化。
在A549/DDP细胞中，转染后ERCC1的mRNA和蛋白质水平降低，对顺铂的敏感性从12.49μg/mL提高到9.27μg/mL。
靶向编码346的ERCC1基因的RNA干扰可增强肺癌细胞A549/DDP对顺铂的敏感性。
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