探讨miR-200b在舌鳞癌细胞中的表达和对增殖与侵袭的影响

发布时间：2018-06-06 04:41

本文选题：miR-200b + 舌鳞癌　；参考：《南华大学》2016年硕士论文

【摘要】：【目的】本研究旨在探讨miR-200b在人舌鳞癌组织和细胞中的表达与临床意义以及生物学意义。【方法】培养正常舌粘膜上皮细胞株(TEC)为对照组(A组),不同舌鳞癌细胞株SCC-9、SCC-2、TSCCa、Tca8113分为B、C、D、E组,作为实验组,运用实时荧光定量PCR检测不同舌鳞癌细胞株与正常舌粘膜上皮细胞株中miR-200b的表达。收集62例舌鳞癌组织与癌旁正常组织,其中癌旁正常组织作为对照组(A1组),舌鳞癌组织为实验组(B1组),运用实时荧光定量PCR检测62例舌鳞癌组织与癌旁正常组织中miR-200b的表达,并分析miR-200b的表达变化与舌鳞癌患者临床病理资料的相关性。运用转染技术将scramble、control-siRNA、miR-200b mimics、Bmi-1-siRNA分别转染于舌鳞癌Tca8113细胞分成A2、A3、B2、B3四组,其中scramble组(A2组)与miR-200b mimics组(B2组),control-siRNA组(A3组)与Bmi-1-siRNA组(B3组)分别配对,A2、A3为对照组,B2、B3为实验组。采用Western blot和qRT-PCR检测miR-200b对Bmi-1基因的蛋白和mRNA表达变化。采用MTT法检测miR-200b与Bmi-1-siRNA对舌鳞癌Tca8113细胞增殖的影响。采用Transwell侵袭实验检测miR-200b与Bmi-1-siRNA对舌鳞癌Tca8113细胞侵袭的影响。【结果】1.qRT-PCR结果显示,与A组比较,B、C、D、E组miR-200b的表达均降低,差异具有统计学意义(P0.05)。与A1组比较,B1组miR-200b的表达亦降低,差异具有统计学意义(P0.05)。相关性分析显示,在62例舌鳞癌组织中,miR-200b的表达水平与患者的临床分期和淋巴结转移呈负相关。2.Western blot与qRT-PCR结果显示,与A2、A3组比较,B2、B3组均能明显下调舌鳞癌Tca8113细胞中Bmi-1的蛋白与mRNA表达水平,差异具有统计学意义(P0.05)。3.MTT结果显示,与A2、A3组比较,B2、B3组OD值均明显减小,差异均有统计学意义(P0.05)。4.Transwell结果显示,与A2、A3组比较,B2、B3组穿过基质胶的Tca8113细胞数明显降低,差异均有统计学意义(P0.01)。【结论】1.miR-200b在舌鳞癌细胞与组织中的表达均下调,miR-200b在舌鳞癌中表达下调与患者的临床分期和淋巴结转移负相关。2.miR-200b可通过靶向Bmi-1抑制舌鳞癌细胞增殖与侵袭。
[Abstract]:[objective] to investigate the expression and clinical significance and biological significance of miR-200b in human tongue squamous cell carcinoma tissues and cells. [methods] normal tongue epithelial cells were cultured as control group A, and different tongue squamous cell lines SCC-9 and SCC-2TSCCaTca8113 were divided into two groups. As an experimental group, real-time quantitative PCR was used to detect the expression of miR-200b in different tongue squamous cell carcinoma cell lines and normal tongue mucosal epithelial cell lines. 62 cases of tongue squamous cell carcinoma and adjacent normal tissues were collected, in which the adjacent normal tissues served as control group A 1 and tongue squamous cell carcinoma as experimental group B 1. The expression of miR-200b in 62 cases of tongue squamous cell carcinoma and adjacent normal tissues was detected by real-time fluorescence quantitative PCR. The correlation between the expression of miR-200b and the clinicopathological data of tongue squamous cell carcinoma was analyzed. Scrambleblebleblecontrol-siRNA-miR-200b mimi-1-siRNA transfected into Tca8113 cells of tongue squamous cell carcinoma was divided into four groups: scramble group (A _ 2 group) and miR-200b mimics group B _ 2 group (control group A _ 3) and Bmi-1-siRNA group (B _ 3 group). Western blot and qRT-PCR were used to detect the expression of Bmi-1 protein and mRNA in miR-200b. The effects of miR-200b and Bmi-1-siRNA on the proliferation of Tca8113 cells in tongue squamous cell carcinoma were detected by MTT assay. Transwell invasion assay was used to detect the effect of miR-200b and Bmi-1-siRNA on the invasion of Tca8113 cells in tongue squamous cell carcinoma. [results] the results of 1.qRT-PCR showed that the expression of miR-200b in group A was significantly lower than that in group A (P 0.05). The expression of miR-200b in group B 1 was significantly lower than that in group A1 (P 0.05). Correlation analysis showed that the expression of miR-200b in 62 cases of tongue squamous cell carcinoma was negatively correlated with the clinical stage and lymph node metastasis. 2. Western blot and qRT-PCR showed that the expression of miR-200b was negatively correlated with the clinical stage and lymph node metastasis in 62 cases of tongue squamous cell carcinoma. The expression of Bmi-1 protein and mRNA in Tca8113 cells of tongue squamous cell carcinoma were significantly down-regulated in B _ 2H _ B _ 3 group compared with A _ 2O _ A3 group. The difference was statistically significant (P 0.05). 3. The results showed that the OD value of B _ (2) B _ (2) B _ (3) group was significantly lower than that of A _ (2) A _ (3) group, and the difference was statistically significant (P 0.05 .4.Transwell). The number of Tca8113 cells passing through matrix glue in B _ 2O _ B _ 3 group was significantly lower than that in A _ 2O _ A _ 3 group. [conclusion] the down-regulation of 1.miR-200b expression in tongue squamous cell carcinoma cells and tissues is negatively correlated with clinical stage and lymph node metastasis. 2. MiR-200b can inhibit tongue squamous cell carcinoma through targeted Bmi-1. Cell proliferation and invasion.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R739.86

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