COX-2基因沉默对Tca8113细胞株成瘤性及侵袭转移能力的影响

发布时间：2018-12-20 07:40

【摘要】：舌鳞状细胞癌是高度转移性并具有鳞状细胞分化潜能的上皮性肿瘤。癌细胞通常会向周围神经血管等正常组织浸润,也会发生颈部淋巴结的转移,淋巴结转移多数情况下在病变发生的同侧区域,但是当病变组织位于中线处或者病变已经累及至中线时,肿瘤细胞则有向对侧区域或者双侧转移的趋势。临床上根据肿瘤大小,部位及转移扩散范围确定TNM分期以明确治疗方案,目前多主张采用手术联合放化疗的综合序列治疗的方式,目前学者们正积极地向个性化基因靶向治疗的方向进行研究。本研究以慢病毒为载体通过RNAi技术转染COX-2基因稳定沉默后对Tca8113细胞株的成瘤性及其侵袭转移能力的影响为目的,初探COX-2基因沉默对人舌鳞状细胞癌作用的可能机制。选取稳定传代的活细胞分别设置COX-2基因沉默组(pLKO.1shCOX-2)、空载体组(Tca8113scr)和未处理组(Tca8113)并对其进行常规细胞培养,采用Western-blot验证COX-2蛋白的沉默效率；采用细胞克隆形成实验检测成瘤能力的变化；通过Transwell小室的侵袭实验和迁移实验检测COX-2基因沉默后侵袭转移能力的变化。通过以上实验研究主要得到了以下结论： (1)慢病毒转染的Tca8113细胞株COX-2蛋白表达明显降低； (2)COX-2基因沉默后可以有效地抑制Tca8113细胞株的成瘤情况； (3)COX-2基因沉默后降低了Tca8113细胞株的侵袭转移能力。
[Abstract]:Tongue squamous cell carcinoma is a highly metastatic epithelial tumor with squamous cell differentiation potential. Cancer cells usually infiltrate into normal tissues such as peripheral nerve and blood vessels, and neck lymph nodes metastasize, most often in the ipsilateral region of the lesion. However, when the lesion tissue is located at the midline or the lesion has been involved in the midline, the tumor cells tend to metastasize to the contralateral region or to both sides. According to the tumor size, location and the range of metastasis and diffusion, TNM staging was determined to determine the treatment plan. At present, it is advocated to adopt the comprehensive sequence therapy of surgery combined with radiotherapy and chemotherapy. At present, scholars are actively conducting research in the direction of individualized gene targeted therapy. The aim of this study was to explore the possible mechanism of COX-2 gene silencing on human tongue squamous cell carcinoma (TSCC) in order to investigate the effect of lentivirus transfection of COX-2 gene on tumorigenesis and invasion and metastasis of Tca8113 cell line by RNAi technique. COX-2 gene silencing group (pLKO.1shCOX-2), empty vector group (Tca8113scr) and untreated group (Tca8113) were selected and cultured by routine cell culture. Western-blot was used to verify the silencing efficiency of COX-2 protein. Cell clone formation assay was used to detect the change of tumorigenic ability, and the invasion and metastasis ability of COX-2 gene after silencing was detected by Transwell chamber invasion test and migration assay. The main conclusions were as follows: (1) the expression of COX-2 protein in lentivirus transfected Tca8113 cell line was significantly decreased; (2) COX-2 gene silencing could effectively inhibit the tumorigenesis of Tca8113 cell line. (3) COX-2 gene silencing decreased the invasion and metastasis ability of Tca8113 cell line.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.86

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