沉默hnRNP A2/B1基因后通过Bcl-2/Bax影响宫颈癌CaSki细胞的凋亡  

Silencing hnRNP A2/B1 induced cell apoptosis via Bcl-2/Bax in cervical cancer CaSki cells

作　　者：蔡僮 许修颖 方文 CAI Tong;XU Xiu-ying;FANG Wen(Department of Clinical Biochemistry, Guizhou Medical University, Guiyang 550004, China)

机构地区：[1]贵州医科大学医学检验学院临床生化教研室

出　　处：《天津医药》2019年第9期902-906,共5页Tianjin Medical Journal

基　　金：国家自然科学基金资助项目（81560481）;贵州省科技厅与贵州医科大学联合基金资助项目（2015-7410）

摘　　要：目的探索沉默核内不均一核糖核蛋白A2/B1(hnRNP A2/B1)基因后对宫颈癌CaSki细胞的增殖、凋亡的影响及其相关作用机制。方法建立稳定沉默hnRNP A2/B1的CaSki细胞株,分为未沉默hnRNP A2/B1的空白组(CaSki组)、转染阴性序列的阴性对照组(CaSki-NC组)、转染阳性hnRNP A2/B1干扰序列的实验组(Ca Ski-shRNA组)。运用实时荧光定量PCR(qRT-PCR)及蛋白印迹法(Western blot)对各组细胞hnRNP A2/B1的mRNA及蛋白表达水平进行验证,四甲基偶氮唑蓝(MTT)法检测各组细胞增殖能力,细胞克隆实验检测各组细胞克隆形成能力,流式细胞术检测各组细胞凋亡情况,Western blot检测各组细胞凋亡相关蛋白Bcl-2、Bax的表达变化。结果与Ca Ski组、CaSki-NC相比较,CaSki-shRNA组细胞hnRNP A2/B1的mRNA及蛋白表达水平均明显减低,细胞的增殖能力在48 h降低,克隆形成能力下降,凋亡率增加,Bcl-2蛋白的表达减低,Bax表达升高(P<0.01),而CaSki组与Ca Ski-NC组之间差异无统计学意义(P>0.05)。结论沉默hnRNP A2/B1基因后能抑制宫颈癌CaSki细胞的增殖能力,且可能通过Bcl-2/Bax影响宫颈癌CaSki细胞的凋亡。Objective To investigate the effect of silencing hnRNP A2/B1 on the proliferation and apoptosis of cervical cancer CaSki cells and the relative mechanism. Methods hnRNP A2/B1 stably silencing CaSki cell line was established and divided into three groups. The cells with hnRNP A2 / B1 target silencing were used as the CaSki-shRNA group, the negative vector was used as the negative control group (CaSki-NC). The blank control group (CaSki) was not treated. Quantitative real-time PCR (qRT-PCR) and Western blot assay were used to detect the mRNA and protein expression levels of hnRNP A2/B1. Cell proliferation and colony formation capacity were determined by MTT assay and plate formation assay. The apoptosis was detected by flow cytometry. Western blot assay was performed to reveal the expressions of apoptosisrelated proteins Bcl-2 and Bax. Results The results of qRT-PCR and Western bolt assay confirmed that the mRNA and protein expressions of hnRNP A2/B1 decreased significantly in the CaSki-shRNA group compared with those of CaSki group and the CaSki-NC group. In the CaSki-shRNA group, the proliferation and colony forming ability of CaSki cells decreased, the apoptosis increased and the expression of Bcl-2 was significantly downregulated and Bax was upregulated (P<0.01).There was no significant difference between CaSki group and CaSki-NC group (P>0.05). Conclusion Silencing hnRNP A2/B1 can inhibit the proliferation in cervical cancer CaSki cells and may induce cell apoptosis via Bcl-2/Bax.

关 键 词：宫颈肿瘤 细胞凋亡 原癌基因蛋白质C-BCL-2 BCL-2相关X蛋白质 核内不均一核糖核蛋白A2/B1 

分 类 号：R711.74[医药卫生—妇产科学]