放疗诱导宫颈癌细胞中经典Wnt通路的表达情况及顺铂耐药性研究

目的:研究放疗诱导后的耐放疗宫颈癌Hela、Siha细胞系顺铂耐药情况及经典Wnt通路相关分子在其化疗耐受中的作用。方法:分割剂量多次照射诱导宫颈癌细胞并成功建立耐放疗细胞系,将细胞分为R0组(空白对照)、R1组(分割剂量4GY,照射6次)和R2组(分割剂量6GY,照射4次)。CCK8法进行耐药实验,Real-time PCR及Western blot法检测Wnt/β-catenin通路及耐药分子表达情况。结果:Hela R0、R1、R2顺铂IC50分别为3.86、6.65μmol/L和6.53μmol/L,Siha R0、Siha R1、Siha R2顺铂IC50分别为4.79、7.25μmol/L和7.98μmol/L;与R0组相比,其他两组的IC50显著升高,差异有统计学意义(P0.001)。放疗诱导宫颈癌Hela、Siha细胞中经典Wnt通路分子在基因与蛋白水平受到激活,其下游基因C-Myc、Cyclin D1表达增多,放疗诱导Hela细胞中耐药蛋白ABCB1、ABCG2表达升高,而放疗诱导Siha细胞中上述耐药蛋白变化无统计学差异。结论:经典Wnt通路可能是放疗诱导宫颈癌细胞化疗耐受性获得的重要机制,这为复发性宫颈癌患者提供了潜在治疗靶点。

The expressions of Wnt canonical pathway and the study of cis-platinum resistance in ra-dio-induced cervical cancer cells

Objective：To investigate the expression of the main molecules in Wnt ca-nonical pathway and cis-platinum resistance in radiation induced cervical cancer cell lines Hela and Siha. Methods：Fractional dose radiotherapy was used to establish radio-resistant cell lines. Real-time PCR and Western Blot assays were employed to test the expressions of the Wnt path-way main members and chemoresistant related protein-ABCB1 and ABCG2. CCK8 assay was used to detect cell viability after cisplatine treatment. Results：The IC50 of R0,R1 and R2 were 3. 86,6. 65,6. 53μmol/ L in Hela,respectively,and 4. 79,7. 25,7. 98μmol/ L in Siha,respec-tively. Compared to R0,IC50 in R1 and R2 were significantly increased（P〈0. 001）. The Wnt canonical pathway proteins such as Wnt1,β-catenin,C-Myc,Cyclin D1 were activated in the ra-dio-resistant cell lines at both mRNA and protein levels,the chemoresistant related protein AB-CB1 and ABCG2 were activated in radio-induced Hela cells only. Conclusions：The activation of Wnt canonical pathway could be a significant molecular mechanism of cis-platinum resistance in radiation induced cervical cancers. It may provide a potential target of treatment for recurrent cervical cancer.