[RNA干扰沉默切除修复交叉互补1基因并影响肺癌细胞系对顺铂的敏感性]
[RNA interference silencing excision repair cross-complementing 1 gene and affecting cisplatin sensitivity on lung cancer cell lines].
作者信息
Shang Xiao-bin, Yu Zhen-tao, Tang Peng, Zhang Xi-zeng
机构信息
Department of Esophageal Tumor, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.
出版信息
Zhonghua Yi Xue Za Zhi. 2008 Oct 28;88(39):2799-802.
OBJECTIVE
To research the changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing (ERCC)1 gene in lung cancer cell lines.
METHODS
The small interference RNA (siRNA) targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to lung cancer cell line A549. 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.
RESULTS
In A549 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfection. The sensitivity to cisplatin of A549 cell line was increased by 3.07 times after disturbing the ERCC1 gene.
CONCLUSIONS
The sensitivity to cisplatin of lung cancer cell lines A549 could be enhanced by RNA interfering ERCC1 gene.
目的
通过RNA干扰肺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。
方法
体外转录设计并合成靶向ERCC1基因的小干扰RNA（siRNA），转染至肺癌细胞系A549。采用逆转录聚合酶链反应（RT-PCR）、蛋白质免疫印迹法（Western blot）和免疫细胞化学法评估ERCC1的信使核糖核酸（mRNA）和蛋白质。通过噻唑蓝（MTT）法检测干扰后顺铂敏感性的变化。
结果
在A549细胞中，转染后24、48和72小时，ERCC1的mRNA和蛋白质水平显著降低。干扰ERCC1基因后，A549细胞系对顺铂的敏感性提高了3.07倍。
结论
RNA干扰ERCC1基因可增强肺癌细胞系A549对顺铂的敏感性。
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