【摘要】：目的：通過觀察高表達(dá)的miR-101對子宮頸癌細(xì)胞增殖、遷移、凋亡等的影響,并分析靶基因蛋白EZH2和COX-2在子宮頸癌細(xì)胞中的表達(dá),探討miR-101在宮頸細(xì)胞癌變中的作用。方法：將人工合成的miR-101的mimics(上調(diào)組)和inhibitor(下調(diào)組)通過LipofectamineTM2000轉(zhuǎn)染實(shí)驗(yàn)組宮頸癌細(xì)胞株(Hela, Siha)以及對照組食管鱗癌細(xì)胞株Eca109。應(yīng)用qRT-PCR險測各組細(xì)胞中miR-101的表達(dá)量、MTT法檢測定細(xì)胞增殖活力、流式細(xì)胞術(shù)檢測細(xì)胞凋亡率、細(xì)胞劃痕實(shí)驗(yàn)觀察細(xì)胞遷移能力,免疫組織化學(xué)法檢測EZH2或COX-2蛋白的表達(dá)。結(jié)果：兩種細(xì)胞株qRT-PCR結(jié)果：上調(diào)組miR-101的表達(dá)量均高于未轉(zhuǎn)染組,下調(diào)組miR-101的表達(dá)量均低于未轉(zhuǎn)染組。MTT結(jié)果：上調(diào)組96后細(xì)胞的增殖能力降低,下調(diào)組96h后細(xì)胞的增殖能力增高。流式細(xì)胞結(jié)果：Hela,Siha細(xì)胞未轉(zhuǎn)染組凋亡率分別為12.2%,8.5%,上調(diào)組凋亡率分別為97.6%,76.6%,下調(diào)組Siha細(xì)胞凋亡率為21.6%。劃痕實(shí)驗(yàn)結(jié)果：Hela上調(diào)組的劃痕距離(42.65um±2um)低于未轉(zhuǎn)染組(181.38um±2um)。Siha上調(diào)組的劃痕距離(36.63um±2um)低于未轉(zhuǎn)染組(184.12um±2um)。免疫組織化學(xué)結(jié)果：上調(diào)組EZH2,COX-2蛋白陽性率較未轉(zhuǎn)染組降低,下調(diào)組兩種蛋白的陽性率較未轉(zhuǎn)染組增高。以上結(jié)果差異均具有統(tǒng)計學(xué)意義(P0.05)。結(jié)論：miR-101的過表達(dá)可抑制子宮頸癌細(xì)胞增殖、抑制宮頸癌細(xì)胞遷移、促進(jìn)癌細(xì)胞的凋亡。miR-101在宮頸細(xì)胞癌變過程中發(fā)揮重要作用。EZH2,COX-2是miR-101的靶基因之一,二者與miR-101之間呈負(fù)性調(diào)節(jié)關(guān)系。
[Abstract]:Aim: to investigate the effects of overexpression of miR-101 on the proliferation, migration and apoptosis of cervical cancer cells, and to analyze the expression of EZH2 and COX-2 in cervical cancer cells, and to explore the role of miR-101 in cervical cancer. Methods: the synthetic miR-101 mimics (up-regulated group) and inhibitor (down-regulated group) were transfected by LipofectamineTM2000 into the cervical cancer cell line (Hela, Siha) and the control esophageal squamous cell carcinoma cell line Eca109.. The expression of miR-101 was measured by qRT-PCR, the proliferative activity was detected by MTT assay, the apoptosis rate was detected by flow cytometry, and the cell migration was observed by cell scratch assay. Immunohistochemical method was used to detect the expression of EZH2 or COX-2 protein. Results: the expression of miR-101 in the up-regulated group was higher than that in the non-transfected group, and the expression of miR-101 in the down-regulated group was lower than that in the untransfected group. MTT results showed that the proliferation ability of the cells in the up-regulated group was decreased after 96 months. The proliferation ability of the down-regulated group was increased after 96 h. Flow cytometry results showed that the apoptotic rate of Hela,Siha cells was 12.28.5in the untransfected group, 97.6and 76.6in the upregulated group, and 21.6in the down-regulated group. The scratch distance (42.65um 鹵2um) of Hela up-regulated group was lower than that of 181.38um 鹵2um). Siha up-regulated group (36.63um 鹵2um) lower than that of untransfected group (184.12um 鹵2um). Immunohistochemical results: the positive rate of EZH2,COX-2 protein in up-regulation group was lower than that in non-transfection group, and the positive rate of two proteins in down-regulated group was higher than that in untransfected group. The differences were statistically significant (P0.05). Conclusion: overexpression of miR-101 can inhibit the proliferation of cervical cancer cells, inhibit the migration of cervical cancer cells and promote the apoptosis of cancer cells. MiR-101 plays an important role in the process of cervical cancer. EZH2,COX-2 is one of the target genes of miR-101. There was a negative regulatory relationship between them and miR-101.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R737.33

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