發(fā)布時(shí)間：2018-03-11 20:15

  本文選題：Snail　切入點(diǎn)：子宮內(nèi)膜癌　出處：《蚌埠醫(yī)學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：第一部分Snail蛋白在子宮內(nèi)膜癌中的表達(dá)與臨床意義 目的:通過(guò)對(duì)Snail在子宮內(nèi)膜癌中表達(dá)的研究來(lái)探索Snail與子宮內(nèi)膜癌浸潤(rùn)和轉(zhuǎn)移及預(yù)后的關(guān)系。 方法:采用免疫組織化學(xué)方法，結(jié)合組織芯片技術(shù)，檢測(cè)124例子宮內(nèi)膜癌、28例不典型增生內(nèi)膜、35例正常內(nèi)膜組織中Snail的表達(dá)水平，，分析Snail蛋白表達(dá)與臨床病理因素的關(guān)系。 結(jié)果:子宮內(nèi)膜癌組織中Snail的表達(dá)率為46.8%，與不典型增生內(nèi)膜和正常內(nèi)膜組織相比，有顯著統(tǒng)計(jì)學(xué)差異(P 0.01)。Snail表達(dá)與FIGO分期、淋巴結(jié)轉(zhuǎn)移明顯相關(guān)(P 0.05)。Snail陽(yáng)性患者生存期明顯短于陰性患者。 結(jié)論：Snail可能通過(guò)調(diào)控HIF-1α和E-cadherin的表達(dá)，在子宮內(nèi)膜癌的發(fā)生及侵襲轉(zhuǎn)移過(guò)程中發(fā)揮重要作用，針對(duì)干預(yù)Snail途徑的方案可能對(duì)于子宮內(nèi)膜癌的診治提供新的方法。 第二部分shRNA抑制Snail基因表達(dá)的作用 目的：通過(guò)shRNA抑制轉(zhuǎn)錄因子Snail的表達(dá)，觀察人子宮內(nèi)膜癌ISK細(xì)胞在體外侵襲轉(zhuǎn)移能力以及和上皮-間質(zhì)轉(zhuǎn)化的關(guān)系。 方法：構(gòu)建表達(dá)Snail的短發(fā)夾RNA(Small hairpin-shaped RNA, shRNA)的RNA干擾載體(Snail shRNA vector)和表達(dá)不針對(duì)任何已知mRNA的shRNA陰性對(duì)照RNA干擾載體(control shRNA vector)，分別轉(zhuǎn)染ISK細(xì)胞后篩選得到Snail表達(dá)受抑制的ISK-shSnail細(xì)胞和Snail表達(dá)未受影響的ISK-shControl細(xì)胞。分別采用RT-PCR和Western blot技術(shù)在mRNA和蛋白水平驗(yàn)證各組細(xì)胞。用CCK-8檢測(cè)各組細(xì)胞的增殖能力。用Transwell小室模型結(jié)合基質(zhì)膠的方法比較各組細(xì)胞在遷移能力和侵襲能力的差異。 結(jié)果：Snail-shRNA組與ISK未轉(zhuǎn)染組相比,Snail表達(dá)明顯減弱(P 0.01),Transwell chamber穿膜細(xì)胞數(shù)顯著減少(P 0.01); ISK-shControl組Snail表達(dá)及Transwell chamber穿膜細(xì)胞數(shù)和ISK未轉(zhuǎn)染組相比無(wú)顯著差異(P0.05)。CCK-8法檢測(cè)細(xì)胞生長(zhǎng)，Snail干擾后ISK細(xì)胞生長(zhǎng)明顯減慢,與未轉(zhuǎn)染組相比有顯著差異。 結(jié)論:通過(guò)RNA干擾沉默Snail基因的表達(dá)能有效地抑制子宮內(nèi)膜癌ISK體外侵襲遷移能力,同時(shí)逆轉(zhuǎn)上皮間質(zhì)轉(zhuǎn)化，針對(duì)干預(yù)Snail途徑的方案可能對(duì)于子宮內(nèi)膜癌的診治具有重要意義。
[Abstract]:The expression and clinical significance of Snail protein in endometrial carcinoma. Objective: to study the expression of Snail in endometrial carcinoma and explore the relationship between Snail and invasion, metastasis and prognosis of endometrial carcinoma. Methods: immunohistochemical method and tissue microarray technique were used to detect the expression of Snail in 28 cases of atypical hyperplasia of endometrial carcinoma and 35 cases of normal endometrium. The relationship between the expression of Snail protein and clinicopathological factors was analyzed. Results: the expression rate of Snail in endometrial carcinoma was 46.8%, which was significantly different from that in atypical hyperplasia endometrium and normal endometrial tissue (P 0.01) and FIGO stage. The survival time of patients with positive lymph node metastasis was significantly shorter than that of patients with negative lymph node metastasis. Conclusion the expression of HIF-1 偽 and E-cadherin may play an important role in the development, invasion and metastasis of endometrial carcinoma by controlling the expression of HIF-1 偽 and E-cadherin. The intervention of Snail pathway may provide a new method for the diagnosis and treatment of endometrial carcinoma. The second part: the role of shRNA in inhibiting the expression of Snail gene. Aim: to investigate the relationship between invasion and metastasis of human endometrial carcinoma ISK cells in vitro and epithelial-interstitial transformation by shRNA inhibiting the expression of transcription factor Snail. Methods: the short hairpin RNA(Small hairpin-shaped (shRNAs) and shRNA negative control RNA interference vector (RNA control shRNA vector) expressing Snail were constructed. After transfection into ISK cells, the expression of Snail was inhibited. ISK-shSnail cells and ISK-shControl cells with unaffected Snail expression were examined by RT-PCR and Western blot techniques at the mRNA and protein levels, respectively. The proliferative ability of each group was detected by CCK-8. The Transwell cell model combined with matrix glue was used to compare the cell proliferation. The difference of cell migration and invasion ability in each group. Results compared with the untransfected ISK group, the expression of Snail in the 10% Snail-shRNA group was significantly lower than that in the untransfected ISK group, while the number of transmembrane cells in the ISK-shControl group and the number of Transwell chamber perforating cells in the ISK-shControl group was significantly decreased compared with the untransfected group of ISK. The growth of ISK cells slowed down after disturbance. There was a significant difference compared with the untransfected group. Conclusion: silencing the expression of Snail gene by RNA interference can effectively inhibit the invasion and migration of endometrial carcinoma ISK in vitro and reverse the epithelial stromal transformation. The intervention of Snail pathway may be of great significance in the diagnosis and treatment of endometrial carcinoma.
【學(xué)位授予單位】：蚌埠醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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