b7 h3 tlt 2在口腔鳞癌中的表达及其意义word格式论文.docxVIP

优秀毕业论文 精品参考文献资料 中文摘要 目的：研究 B7-H3、TLT-2 在口腔鳞癌组织和正常口腔黏膜组织中的表达差异，以及 B7-H3 对口腔鳞癌细胞生物学的影响及其意义。 方法：采用普通 PCR、实时荧光定量 PCR 和免疫组化检测 B7-H3、TLT-2 在口腔鳞癌组 织和正常口腔黏膜组织的表达差异；构建 B7-H3 腺病毒表达载体，感染人舌鳞癌细胞株 Tca8113，荧光显微镜下观测 GFP 荧光的强度和密度，RT-qPCR 检测各组细胞内 B7-H3mRNA 表达量，以确定 Adv-pEGFP-B7-H3 表达载体的感染效率。再采用 CCK-8、 PI 染色流式细胞仪检测 B7-H3 高表达对 Tca8113 细胞增殖和细胞周期的影响。采用 SPSS11.0 软件包对数据进行处理。 结果：普通 PCR 显示，口腔鳞癌组织及正常口腔黏膜组织均表达 B7-H3 和 TLT-2mRNA， 但 RT-qPCR 结果显示目的基因在两者中表达差异较大，口腔鳞癌组织 B7-H3mRNA 表达 水平相当于正常口腔粘膜的 7.2904±0.3647 倍，而口腔鳞癌组织 TLT-2mRNA 表达水平相 当于正常口腔粘膜的 5.8573±0.3504 倍，均显著高于正常口腔黏膜组织(P0.05)；与对照 组相比，感染复数(MOI)为 10、50、100 组在作用 24h 呈现促进细胞增殖作用，S 期细胞 比例显著增加，在 72h 时增殖作用最强(P0.05)；与感染复数(MOI)1 组比较，同时间段 50、100 组促细胞增殖作用和 S 期细胞比例显著增加(P0.05)，MOI 50 组与 MOI 100 组 比较无显著差异(P0.05)。 结论：口腔鳞癌组织B7-H3、TLT-2mRNA和蛋白表达显著高于其在正常口腔黏膜组织中表 达；在人舌鳞癌细胞株Tca8113中，B7-H3高表达可促进口腔鳞癌细胞增殖。 关键词：口腔鳞癌; Tca8113细胞; B7-H3; TLT-2; 细胞增殖 II Abstract PURPOSE: To investigate the differences of B7-H3 and TLT-2 expression between in oral squamous cell carcinoma tissues and normal oral mucosa, then further research the function and significance of B7-H3 expression on the biology of oral squamous cell carcinoma cells. METHODS: B7-H3 and TLT-2 expression in oral squamous cell carcinoma tissues and normal oral mucosa were detected by RT-PCR/RT-qPCR and immunohistochemical assay. B7-H3 adenovirus expression vector was constructed, and Tca8113 cells were transfected. Then we have used fluorescence microscope to observe GFP fluorescence intensity and density, in order to detect infection efficiency of Adv-pEGFP-B7-H3. B7-H3mRNA expression in each group cells were detected by RT-qPCR, then the cell proliferation and cell cycle were evaluated by CCK-8 and flow cytometry (PI staining) assay, respectively. The date was analyzed with SPSS 11.0 software package. RESULTS: RT-PCR results showed B7-H3 and TLT-2mRNA are both expressed in oral squamous cell carcinoma and normal oral mucosa. But RT-qPCR results showed that purpose gene have difference expression between two groups.