宫颈癌细胞系Siha和C33a中RRM2表达水平与细胞对吉西他滨敏感性的相关分析

Correlation between RRM2 Expression Level of Cervical Cancer Cell Line Siha and C33a and Cell Sensitivity to Gemcitabine

目的：研究宫颈癌细胞系C33a和Siha中核糖核苷酸还原酶小亚基M2（RRM2）表达与细胞对吉西他滨（Gem）敏感性的相关性。方法：以0.1-3.2μmol/L Gem处理C33a细胞,以0.5-512μmol/L Gem处理Siha细胞,72 h后用MTT法测定细胞活力,计算半数抑制浓度（IC50）;使用Western blot法和逆转录-聚合酶链反应检测各细胞中RRM2表达;采用浓度梯度倍增和大剂量冲击相结合的方法对Siha细胞进行Gem耐药诱导培养,构建Siha/Gem耐药细胞株;在Siha/Gem细胞中,使用RNA干扰技术敲低RRM2表达并检测敲低前后Gem对细胞的IC50。结果：Gem对C33a、Siha细胞的IC50分别为0.63、16.8μmol/L。与C33a细胞比较,Siha细胞中RRM2的表达较高。与Siha细胞相比,Siha/Gem耐药细胞（耐药指数为16.26）中RRM2表达增强。Gem对敲低RRM2表达的Siha/Gem耐药细胞的IC50降低,其逆转耐药倍数为4.24。结论：宫颈癌细胞系中RRM2表达可能与细胞对Gem的敏感性呈负相关,敲低RRM2表达可增加细胞对Gem的敏感性。

OBJECTIVE：To study the correlation between RRM2 expression of cervical cancer cell line C33 a and Siha and cell sensitivity to gemcitabine（Gem）. METHODS：C33a and Siha were treated with 0.1-3.2 μmol/L and 0.5-512 μmol/L of gemcitabine respectively for 72 h;cell viability was measured by MTT assay to calculate the value of IC50. The expression of RRM2 was measured by Western blot and RT-PCR. Siha cell was treated with gradient concentration and large dose of Gem to establish Siha/Gem drug-resistant cell line. RNA interference technology knockdown the expression of RRM2 in Siha/Gem cell,and IC50 of Gem to cell was determined before and after knockdown. RESULTS：The IC50 values of Gem to Siha and C33 a were 16.8 μ mol/L and 0.63μmol/L. Compared with C33 a cells,the expression of RRM2 in Siha cell was higher. Compared with Siha cells,Siha/Gem drug-resistant cell（drug resistant index of 16.26）showed higher RRM2 expression. Siha/Gem drug-resistant cell knockdown RRM2,IC50 of Gem to it was decreased,and inverse drug resistant times was 4.24. CONCLUSIONS：There is an negative correlation between RRM2 expression and Gem sensitivity in cervical cancer cell lines. The knockdown of RRM2 in Siha/Gem increases the sensitivity to Gem.