摘要
目的探讨MEX3A基因沉默对膀胱癌细胞增殖和凋亡的影响,并验证MEX3A蛋白在人膀胱癌组织中的表达。方法选取8例病理诊断证实为膀胱癌患者的癌组织和癌周组织标本,其中男性5例,女性3例;年龄50~84岁,平均年龄63.4岁。实验组应用MEX3A-shRNA慢病毒颗粒转染5637膀胱癌细胞株,对照组采用阴性对照慢病毒转染。实时定量聚合酶链反应(PCR)检测MEX3A基因敲减效率,使用噻唑蓝(MTT)法检测细胞增殖,Caspase3/7法检测细胞凋亡。应用免疫组织化学染色检测膀胱癌及癌周组织细胞中MEX3A蛋白表达量。结果实时荧光定量PCR检测经shRNA慢病毒感染两组目的基因mRNA水平的表达丰度,实验组为(0.26±0.05),对照组为(1.00±0.05);两组比较,MEX3A基因在实验组5637膀胱癌细胞中的mRNA水平的表达量受到抑制(t=17.7,P<0.05)。实验组与对照组相比,细胞增殖明显受到抑制(P<0.05),细胞凋亡显著增加(P<0.05)。在400倍光学显微镜下观察,棕黄色颗粒为人MEX3A蛋白与兔MEX3A抗体反应后染色所致,表示有MEX3A蛋白表达;在癌组织中较多,着色较深,癌周组织中较少,着色较浅。MEX3A蛋白在癌及癌周组织总的表达量分别为(7.33±2.73)分、(4.42±1.98)分,两组比较差异有统计学意义(P<0.05);癌组织中的表达量约是癌周组织中的1.66倍。结论MEX3A基因可以促进膀胱癌细胞增殖,并抑制其凋亡;MEX3A蛋白在膀胱癌组织中呈现高表达。
Objective To investigate the effect of MEX3A reticence on proliferation and apoptosis of bladder cancer cells, and verify the expression of MEX3A protein in human bladder cancer. Methods The cancer and peri-cancerous tissue samples of 8 patients who were pathologically proved as bladder cancer were collected, which included 5 males and 3 females, aged 50-84 years old with mean age of 63.4 years old. The 5637 bladder cancer cell in experimental group was transfected with MEX3 A-shRNA lentiviral particles, and control group transfected with negative control lentivirus. The knockdown efficiency of MEX3A gene was detected by real-time polymerase chain reaction（PCR）, and cell proliferation was detected by methylthia-zolyldiphenyl-tetrazolium bromide（MTT） assay, and apoptosis was detected Caspase 3/7 assay. The cancer and peri-cancerous tissues were observed by immunohistochemistry respectively, and then detected expression levels of MEX3A protein.Results The expression of shRNA lentivirus infected mRNA levels of 2 groups were detected by real-time quantitative PCR,experimental group was（0.26 ± 0.05） and control group was（1.00 ± 0.05）. The mRNA level expression of MEX3A in 5637 bladder cancer cells of experimental group was inhibited（t = 17.7, P〈0.05）. Compared with control group, the cell proliferation was significantly inhibited in experimental group（P〈0.05）, while apoptosis was significantly increased（P〈0.05）. Observed under 400-fold optical microscope, the brown granules was stained reaction product of human MEX3A protein and rabbit MEX3A antibody, which showed MEX3A protein expression. The granules distributed in cancer tissue with darker color and less were observed in peri-cancerous tissue with lighter color. The total expressed levels of MEX3A protein in cancer and peri-cancerous tissues were（7.33 ± 2.73） scores and（4.42 ± 1.98） scores, and there were statistical significances between 2 groups（P〈0.05）, and the expression levels of MEX3A protein in cancer was 1.66 times of that in peri-cancerous tissues. Conclusion It is demonstrated that the MEX3A gene may promote proliferation of bladder cancer cells and inhibit its apoptosis, and MEX3A protein shows high expression in bladder cancer tissue.
出处
《生物医学工程与临床》
CAS
2018年第1期82-86,共5页
Biomedical Engineering and Clinical Medicine
基金
国家自然科学基金资助项目(81371552)
