miR-124在卵巢癌中的表达及其对细胞生物学行为的影响

发布时间：2018-05-18 18:53

本文选题：卵巢癌 + miR-124　；参考：《重庆医科大学》2014年硕士论文

【摘要】：目的：卵巢癌发病率位居妇科恶性肿瘤第三位，，由于缺乏特异性症状，早期诊断困难，而晚期卵巢癌的死亡率却高居不下。因此，研究的热点致力于发现卵巢癌的病因及危险因素，并探索可用于早期诊断卵巢癌及预示预后的肿瘤标志物。本文拟探讨miR-124在卵巢癌组织中的表达及其对卵巢癌细胞生物学特性的影响。方法：收集2008年5月至2012年11月期间重庆市肿瘤研究所织标本库有完整病历资料的35例卵巢癌患者组织标本及对应的癌旁组织，利用实时荧光定量-PCR方法检测卵巢癌组织中miR-124的表达水平；利用脂质体转染法分别将miR-124模拟片段(mimic)转染入卵巢癌SKOV3细胞中，恢复细胞内miR-124表达；分别运用CCK-8和流式细胞术检测miR-124对卵巢癌SKOV3细胞增殖及凋亡的影响；transwell小室法检测其对SKOV3细胞迁移及侵袭能力的影响；运用western blot法检测SKOV3细胞中PIK3CA及其下游蛋白的表达量变化。结果：35例卵巢癌患者肿瘤组织中miR-124表达量(1.5254±1.7054)与对应癌旁组织表达量(2.9420±3.4422)相比明显下降，差异具有统计学意义(P＜0.05)；通过转染miR-124模拟片段恢复了卵巢癌SKOV3细胞中miR-124的表达；与lipofectamine组和negative control组相比，转染miR-124组卵巢癌细胞增殖能力下降(P＜0.01)，早期凋亡比例增加(P＜0.05)，侵袭(P＜0.01)和迁移能力下降(P＜0.01)；并且过表达miR-124的SKOV3细胞PIK3CA及下游的p-AKT表达明显下降(P＜0.01)，而总AKT蛋白无明显变化（P＞0.05）。结论：miR-124在卵巢癌组织中表达量下降，且恢复其在卵巢癌细胞中的表达能够明显抑制其细胞增殖、迁移和侵袭，促进细胞凋亡，这一抑制功能可能是部分通过抑制卵巢癌细胞中PI3K/AKT信号通路的活性产生的。
[Abstract]:Objective: the incidence of ovarian cancer is the third in gynecological malignant tumors, and the early diagnosis is difficult because of lack of specific symptoms, while the mortality rate of advanced ovarian cancer is high. Therefore, the focus of the study is to identify the etiology and risk factors of ovarian cancer, and to explore tumor markers for early diagnosis and prognosis of ovarian cancer. The purpose of this study was to investigate the expression of miR-124 in ovarian cancer tissues and its effect on the biological characteristics of ovarian cancer cells. Methods: from May 2008 to November 2012, 35 specimens of ovarian cancer tissues and their corresponding adjacent tissues were collected from Chongqing Institute of Oncology (Chongqing Institute of Oncology) with complete medical records. The expression of miR-124 in ovarian cancer tissues was detected by real-time fluorescence quantification-PCR method, and the mimic miR-124 fragment was transfected into ovarian cancer SKOV3 cells by liposome transfection to restore the expression of miR-124. Effects of miR-124 on proliferation and apoptosis of ovarian cancer SKOV3 cells were detected by CCK-8 and flow cytometry respectively. The effects of miR-124 on the migration and invasion of SKOV3 cells were detected by transwell chamber assay, and the expression of PIK3CA and its downstream proteins in SKOV3 cells were detected by western blot assay. Results the expression of miR-124 in the tumor tissues of 35 patients with ovarian cancer was significantly lower than that in the corresponding paracancerous tissues (2.9420 卤3.4422) (P < 0.05), and the expression of miR-124 in ovarian cancer SKOV3 cells was recovered by transfection of miR-124 mimic fragments. Compared with the lipofectamine and negative control groups, In miR-124 transfection group, the proliferative ability of ovarian cancer cells decreased (P < 0.01), the proportion of early apoptosis increased (P < 0.05), invasion and migration decreased (P < 0.01), and the expression of PIK3CA and downstream p-AKT in SKOV3 cells with overexpression of miR-124 decreased significantly (P < 0.01), but the total AKT protein did not change significantly (P > 0.05). Conclusion the cell proliferation, migration, invasion and apoptosis of ovarian cancer cells can be significantly inhibited by the expression of 1% miR-124 in ovarian cancer cells, and the recovery of the expression in ovarian cancer cells can promote the apoptosis of ovarian cancer cells. This inhibition may be partly due to inhibition of PI3K/AKT signaling pathway activity in ovarian cancer cells.
【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】：R737.31

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