摘要
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.
出处
《中国药房》
CAS
北大核心
2015年第34期4792-4794,共3页
China Pharmacy