miR-369-3p通过调控VEGFC基因表达对膀胱癌细胞增殖及凋亡的影响  

Effects of miR-369-3p on the proliferation and apoptosis of bladder cancer cells by regulating the expression of VEGFC gene

作　　者：赵振伶 邵焕军[1] 郝丽娜 刘军[1] 

机构地区：[1]安徽省亳州市人民医院泌尿外科,236800 [2]昆明医科大学第一附属医院泌尿外科,650032

出　　处：《中国医师杂志》2018年第1期92-95,99,共5页Journal of Chinese Physician

摘　　要：目的观察miR-369-3p对膀胱癌细胞EJ和5637中血管内皮细胞生长因子C(VEGFC)基因的调控作用,并观察其对细胞增殖及凋亡的影响。方法转染miR-NC(对照组)或转染miR-369-3p(观察组)至膀胱癌细胞株EJ和5637;采用Targetscan靶基因预测软件预测miR-369-3p的靶基因;采用荧光实时定量聚合酶链反应(qPCR)检测VEGFC mRNA表达变化;Western blotting检测VEGFC、p-Raf-1和磷酸化细胞外信号调节激酶1/2(p-ERK1/2)在蛋白水平的变化;采用细胞计数试剂盒(CCK-8法)和克隆形成实验检测miR-369-3p对膀胱癌细胞增殖的影响;采用流式细胞术检测miR-369-3p对细胞凋亡的影响。结果转染miR-369-3p后,EJ、5637细胞中VEGFC在mRNA水平和蛋白水平的表达均明显下降(P<0.05),p-Raf-1和p-ERK1/2蛋白表达明显降低,细胞的增殖能力明显降低(P<0.05),细胞形成的克隆数明显减少(P<0.05),细胞凋亡明显增加(P<0.01)。结论 miR-369-3p可通过干扰VEGFC基因的表达,抑制膀胱癌细胞的增殖及促进膀胱癌细胞的凋亡,可能为膀胱癌的生物治疗提供一个新的靶标。Objective To observe the effect of miR-369-3p on vascular endothelial growth factor C （VEGFC） gene in bladder cancer cells EJ and 5637 and observe its effect on cell proliferation and apopto- sis. Methods Bladder cancer cell lines EJ and 5637 were transfected with miR-NC （control group） or transfected with miR-369-3p （experimental group）. The target gene of miR-369-3p was predicted by Tar- gets can target gene prediction software. Fluorescence real-time quantitative polymerase chain reaction （qPCR） was used to detect the changes of VEGFC at mRNA level. The changes of VEGFC, p-Raf-1 and phospborylated extracellular signal-regulated kinase 1/2 （p-ERK1/2） at the protein level were detected by Western blotting. The effect of miR-369-3p on the proliferation of bladder cancer cells was detected by cell counting kit （ CCK-8 ） and clone formation experiment. The effect of miR-369-3p on apoptosis was detected by flow cytometry. Results After transfection with miR-369-3p, the expression of VEGFC at mRNA and protein levels was significantly decreased （ P 〈 0. 05 ） , the expression of p-Raf-1 and p-ERK1/2 protein was significantly decreased, the cell proliferation ability was significantly decreased （ P 〈 0. 05 ） , the number of clones formed was significantly decreased （ P 〈 0. 05 ） and the apoptosis was significantly increased （ P 〈 O. 01 ）. Conclusions miR-369-3p can inhibit the proliferation and promote the apoptosis of bladder cancer cells by interfering with the expression of VEGFC gene, which may provide a new target for the biological therapy of bladder cancer.

关 键 词：微RNAs/药理学 血管内皮生长因子c/药物作用/代谢 膀胱肿瘤/药物疗法/病 理学/代谢 细胞增殖/药物作用 细胞凋亡/药物作用 

分 类 号：R737.14[医药卫生—肿瘤]