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  本文選題：Exosomes　切入點：MicroRNA-29c　出處：《重慶醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：研究膀胱癌細胞分泌的exosomes源性microRNA-29c是否通過下調BCL-2及MCL-1誘導膀胱癌細胞凋亡。 材料與方法：免疫組織化學檢測28例膀胱癌及其癌旁標本BCL-2，MCL-1的表達。實時熒光定量PCR檢測膀胱癌組織及癌旁標本中microRNA-29c的表達。將攜帶microRNA-29c的腺病毒感染膀胱癌BIU-87細胞，超濾和蔗糖密度梯度法提取膀胱癌源性exosomes。實時熒光定量PCR檢測細胞上清液中exosomes中microRNA-29c表達，將帶有microRNA-29c的exosomes、空載組及空白組exosomes作用于BIU-87細胞，流式細胞儀檢測凋亡結果，RT-PCR及western blot檢測BCL-2及MCL-1變化。 結果：膀胱癌及癌旁組織中BCL-2表達陽性率分別為60.7%、17.9%（P=0.01），MCL-1為82.1%、10.7%（P0.01），組間差異有統(tǒng)計學意義；膀胱癌組織中BCL-2與MCL-1表達與microRNA表達呈負相關。經microRNA-29c腺病毒感染的膀胱癌細胞分泌的exosomes中microRNA-29c表達水平為9.97±2.73，空白組及空載組分別為1.00±0.15、1.57±0.35，，組間差異有統(tǒng)計學意義(P0.05)；exosomes源性microRNA-29c作用于膀胱癌細胞后凋亡率（27.77±1.30）%明顯高于空白（3.47±0.81）%和空載組（1.53±0.25）%，組間差異有統(tǒng)計學意義(P0.01)，同時能降低BCL-2mRNA和蛋白表達及MCL-1蛋白表達。 結論：經microRNA-29c腺病毒感染的膀胱癌細胞能通過exosomes運輸microRNA-29c，exosomes源性microRNA-29c可通過下調BCL-2及MCL-1蛋白表達，誘導膀胱癌細胞凋亡。
[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 鹵2.73, and that in blank group and no-load group was 1.00 鹵0.15 ~ 1.57 鹵0.35, respectively. The difference between the two groups was statistically significant. The apoptotic rate was 27.77 鹵1.30% in bladder cancer cells, which was significantly higher than that in blank group (3.47 鹵0.81%) and no-load group (1.53 鹵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.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.14

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