Exosomes源性microRNA-29c诱导膀胱癌细胞凋亡的研究

发布时间：2018-03-14 06:35

本文选题：Exosomes　切入点：MicroRNA-29c　出处：《重庆医科大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：研究膀胱癌细胞分泌的exosomes源性microRNA-29c是否通过下调BCL-2及MCL-1诱导膀胱癌细胞凋亡。材料与方法：免疫组织化学检测28例膀胱癌及其癌旁标本BCL-2，MCL-1的表达。实时荧光定量PCR检测膀胱癌组织及癌旁标本中microRNA-29c的表达。将携带microRNA-29c的腺病毒感染膀胱癌BIU-87细胞，超滤和蔗糖密度梯度法提取膀胱癌源性exosomes。实时荧光定量PCR检测细胞上清液中exosomes中microRNA-29c表达，将带有microRNA-29c的exosomes、空载组及空白组exosomes作用于BIU-87细胞，流式细胞仪检测凋亡结果，RT-PCR及western blot检测BCL-2及MCL-1变化。结果：膀胱癌及癌旁组织中BCL-2表达阳性率分别为60.7%、17.9%（P=0.01），MCL-1为82.1%、10.7%（P0.01），组间差异有统计学意义；膀胱癌组织中BCL-2与MCL-1表达与microRNA表达呈负相关。经microRNA-29c腺病毒感染的膀胱癌细胞分泌的exosomes中microRNA-29c表达水平为9.97±2.73，空白组及空载组分别为1.00±0.15、1.57±0.35，，组间差异有统计学意义(P0.05)；exosomes源性microRNA-29c作用于膀胱癌细胞后凋亡率（27.77±1.30）%明显高于空白（3.47±0.81）%和空载组（1.53±0.25）%，组间差异有统计学意义(P0.01)，同时能降低BCL-2mRNA和蛋白表达及MCL-1蛋白表达。结论：经microRNA-29c腺病毒感染的膀胱癌细胞能通过exosomes运输microRNA-29c，exosomes源性microRNA-29c可通过下调BCL-2及MCL-1蛋白表达，诱导膀胱癌细胞凋亡。
[Abstract]:Aim: to investigate whether exosomes derived microRNA-29c secreted by bladder cancer cells can induce apoptosis of bladder cancer cells by down-regulating BCL-2 and MCL-1. Materials and methods: the expression of BCL-2mCL-1 in 28 cases of bladder cancer and its adjacent specimens was detected by immunohistochemistry. The expression of microRNA-29c in bladder cancer tissues and adjacent specimens was detected by real-time fluorescence quantitative PCR. The adenovirus carrying microRNA-29c was infected into BIU-87 cells of bladder cancer. Ultrafiltration and sucrose density gradient method were used to extract exosomes from bladder carcinomas. The expression of microRNA-29c in the supernatant of cells was detected by real-time fluorescence quantitative PCR. The exosomes with microRNA-29c, exosomes in empty and blank groups were used in BIU-87 cells. Apoptosis was detected by flow cytometry and BCL-2 and MCL-1 were detected by western blot and RT-PCR. Results: the positive rates of BCL-2 expression in bladder cancer and adjacent tissues were 60.7 and 17.9, respectively. The MCL-1 of P0.01 and MCL-1 were 82.1 and 10.7, respectively. The difference between the two groups was statistically significant. There was a negative correlation between the expression of BCL-2 and MCL-1 and the expression of microRNA in bladder cancer tissues. The expression of microRNA-29c in exosomes secreted by microRNA-29c adenovirus infected bladder cancer cells was 9.97 卤2.73, and that in blank group and no-load group was 1.00 卤0.15 ~ 1.57 卤0.35, respectively. The difference between the two groups was statistically significant. The apoptotic rate was 27.77 卤1.30% in bladder cancer cells, which was significantly higher than that in blank group (3.47 卤0.81%) and no-load group (1.53 卤0.25%). The difference was statistically significant (P 0.01). The expression of BCL-2mRNA and protein and the expression of MCL-1 protein were also decreased. Conclusion: bladder cancer cells infected with microRNA-29c adenovirus can transport microRNA-29cTexosomes derived microRNA-29c through exosomes and induce apoptosis by down-regulating the expression of BCL-2 and MCL-1 proteins.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.14

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