干扰Snail基因表达对子宫内膜癌侵袭转移的影响

发布时间：2018-03-11 20:15

本文选题：Snail　切入点：子宫内膜癌　出处：《蚌埠医学院》2014年硕士论文　论文类型：学位论文

【摘要】：第一部分Snail蛋白在子宫内膜癌中的表达与临床意义目的:通过对Snail在子宫内膜癌中表达的研究来探索Snail与子宫内膜癌浸润和转移及预后的关系。方法:采用免疫组织化学方法，结合组织芯片技术，检测124例子宫内膜癌、28例不典型增生内膜、35例正常内膜组织中Snail的表达水平，，分析Snail蛋白表达与临床病理因素的关系。结果:子宫内膜癌组织中Snail的表达率为46.8%，与不典型增生内膜和正常内膜组织相比，有显著统计学差异(P 0.01)。Snail表达与FIGO分期、淋巴结转移明显相关(P 0.05)。Snail阳性患者生存期明显短于阴性患者。结论：Snail可能通过调控HIF-1α和E-cadherin的表达，在子宫内膜癌的发生及侵袭转移过程中发挥重要作用，针对干预Snail途径的方案可能对于子宫内膜癌的诊治提供新的方法。第二部分shRNA抑制Snail基因表达的作用目的：通过shRNA抑制转录因子Snail的表达，观察人子宫内膜癌ISK细胞在体外侵袭转移能力以及和上皮-间质转化的关系。方法：构建表达Snail的短发夹RNA(Small hairpin-shaped RNA, shRNA)的RNA干扰载体(Snail shRNA vector)和表达不针对任何已知mRNA的shRNA阴性对照RNA干扰载体(control shRNA vector)，分别转染ISK细胞后筛选得到Snail表达受抑制的ISK-shSnail细胞和Snail表达未受影响的ISK-shControl细胞。分别采用RT-PCR和Western blot技术在mRNA和蛋白水平验证各组细胞。用CCK-8检测各组细胞的增殖能力。用Transwell小室模型结合基质胶的方法比较各组细胞在迁移能力和侵袭能力的差异。结果：Snail-shRNA组与ISK未转染组相比,Snail表达明显减弱(P 0.01),Transwell chamber穿膜细胞数显著减少(P 0.01); ISK-shControl组Snail表达及Transwell chamber穿膜细胞数和ISK未转染组相比无显著差异(P0.05)。CCK-8法检测细胞生长，Snail干扰后ISK细胞生长明显减慢,与未转染组相比有显著差异。结论:通过RNA干扰沉默Snail基因的表达能有效地抑制子宫内膜癌ISK体外侵袭迁移能力,同时逆转上皮间质转化，针对干预Snail途径的方案可能对于子宫内膜癌的诊治具有重要意义。
[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 伪 and E-cadherin may play an important role in the development, invasion and metastasis of endometrial carcinoma by controlling the expression of HIF-1 伪 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.
【学位授予单位】：蚌埠医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

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