RNPC1基因对乳腺癌细胞MCF-7阿霉素药物敏感性的影响

Effects of RNPC1 on the sensitivity of breast cancer cell MCF-7 with doxorubicin

目的 :研究RNPC1对乳腺癌细胞MCF-7阿霉素药物敏感性的影响。方法 :在MCF-7细胞中使用慢病毒转染法,设敲除(sh RNPC1)组和过表达RNPC1组、敲除对照(SCR)组和过表达对照(NC)组。用实时荧光定量PCR和Western blot法检测各组MCF-7细胞中RNPC1的m RNA和蛋白水平。以不同浓度的阿霉素处理各组MCF-7细胞,CCK-8法检测各组细胞生长抑制率。阿霉素处理各组MCF-7细胞后,用流式细胞术检测各组细胞凋亡率,Western blot法检测各组细胞中调亡相关蛋白的表达。结果:sh RNPC1组RNPC1 m RNA和蛋白相对表达量均低于SCR组,过表达RNPC1组RNPC1 m RNA及蛋白相对表达量均高于NC组(P<0.05)。阿霉素处理各组MCF-7细胞后,sh RNPC1组IC50明显于低于SCR组(P<0.05),过表达组IC50明显高于NC组(P<0.05)。阿霉素处理各组MCF-7细胞24 h后,发现sh RNPC1组凋亡率明显高于SCR组;过表达RNPC1组凋亡率显著低于NC组(P<0.05)。阿霉素处理各组MCF-7细胞24 h后,sh RNPC1组抑凋亡蛋白BCL-XL、BCL-2的表达低于SCR组,促凋亡蛋白BIM和BID的表达高于SCR组;过表达RNPC1组抑凋亡蛋白BCL-CL、BCL-2的表达高于NC组,促凋亡蛋白BIM和BID的表达低于NC组。结论:RNPC1能降低乳腺癌细胞MCF-7对阿霉素的敏感性,其机制与抗细胞凋亡有关。

Objective： To investigate the effects of RNPC1 on the sensitivity of breast cancer cell MCF-7 with doxorubicin （DOX）.Methods： Lentivirus was used to over-express and knock-down RNPC1 in the MCF-7 breast cancer cells.The cells were divided into overexpress RNPC1 （RNPC1） group and its control （NC group）,knock-down RNPC1 （sh RNPC1 group） and its control （SCR group）.The relative m RNA and protein expression of RNPC1 was detected by q RT-PCR and Western blot,respectively.The experimental groups were treated with different concentration of DOX,and cell growth inhibition rate was tested cell counting kit 8 （CCK-8）.After the experimental groups was treated with DOX,cell apoptosis rate was analysis by flow cytometer assay and the expression of apoptosis-related protein was detected by Western blot assay.Results： The q RT-PCR and Western blot results showed that the m RNA and protein level of RNPC1 was increased after transfection with RNPC1 overexpression （RNPC1）.While,RNPC1 was reduced after transfection with knockdown RNPC1 （sh RNPC1） lentivirus.After treatment with different concentration of DOX,the IC50 of sh RNPC1 group was significantly lower than SCR group （P 0.05）.Furthermore,the IC50 of RNPC1 group was higher than NC group （P0.05）.The flow cytometer assay was used after treatment with DOX for 24 h in the experimental groups.The cell apoptosis rate of sh RNPC1 group was higher than SCR group,while,the cell apoptosis rate of RNPC1 was lower than NC group.Overexpression of RNPC1 increased the expression of BCL-XL and BCL-2 after treatment with DOX.Reversely,knockdown of RNPC1 reduced the expression of BCL-XL and BCL-2.Furthermore,overexpression of RNPC1 reduced the expression of BIM and BID after treatment with DOX.While,knockdown of RNPC1 increased the expression of BIM and BID.Conclusion： RNPC1 could decrease the sensitivity of breast cancer cell MCF-7to DOX,and the underlying mechanism might be related to cell apoptosis.