本文選題：卵巢癌 + miR-124�。� 參考：《重慶醫(yī)科大學》2014年碩士論文

【摘要】：目的：卵巢癌發(fā)病率位居婦科惡性腫瘤第三位，，由于缺乏特異性癥狀，早期診斷困難，而晚期卵巢癌的死亡率卻高居不下。因此，研究的熱點致力于發(fā)現(xiàn)卵巢癌的病因及危險因素，并探索可用于早期診斷卵巢癌及預示預后的腫瘤標志物。本文擬探討miR-124在卵巢癌組織中的表達及其對卵巢癌細胞生物學特性的影響。 方法：收集2008年5月至2012年11月期間重慶市腫瘤研究所織標本庫有完整病歷資料的35例卵巢癌患者組織標本及對應的癌旁組織，利用實時熒光定量-PCR方法檢測卵巢癌組織中miR-124的表達水平；利用脂質(zhì)體轉(zhuǎn)染法分別將miR-124模擬片段(mimic)轉(zhuǎn)染入卵巢癌SKOV3細胞中，恢復細胞內(nèi)miR-124表達；分別運用CCK-8和流式細胞術(shù)檢測miR-124對卵巢癌SKOV3細胞增殖及凋亡的影響；transwell小室法檢測其對SKOV3細胞遷移及侵襲能力的影響；運用western blot法檢測SKOV3細胞中PIK3CA及其下游蛋白的表達量變化。 結(jié)果：35例卵巢癌患者腫瘤組織中miR-124表達量(1.5254±1.7054)與對應癌旁組織表達量(2.9420±3.4422)相比明顯下降，差異具有統(tǒng)計學意義(P＜0.05)；通過轉(zhuǎn)染miR-124模擬片段恢復了卵巢癌SKOV3細胞中miR-124的表達；與lipofectamine組和negative control組相比，轉(zhuǎn)染miR-124組卵巢癌細胞增殖能力下降(P＜0.01)，早期凋亡比例增加(P＜0.05)，侵襲(P＜0.01)和遷移能力下降(P＜0.01)；并且過表達miR-124的SKOV3細胞PIK3CA及下游的p-AKT表達明顯下降(P＜0.01)，而總AKT蛋白無明顯變化（P＞0.05）。 結(jié)論：miR-124在卵巢癌組織中表達量下降，且恢復其在卵巢癌細胞中的表達能夠明顯抑制其細胞增殖、遷移和侵襲，促進細胞凋亡，這一抑制功能可能是部分通過抑制卵巢癌細胞中PI3K/AKT信號通路的活性產(chǎn)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 鹵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.
【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】：R737.31

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