端粒酶卡侯体蛋白1干扰慢病毒对口腔鳞状细胞癌细胞裸鼠致瘤作用的研究

发布时间：2019-05-27 12:05

【摘要】：目的探测端粒酶卡侯体蛋白1(TCAB1)基因沉默对裸鼠移植瘤生长及肿瘤细胞生物学行为的影响。方法采用定量荧光原位杂交(Q-FISH)方法测定RNA干扰慢病毒sh TCAB1和sh阴性对照(NC)处理后经稳定筛选26代人口腔鳞状细胞癌(OSCC)的Cal27细胞系染色体末端端粒的信号强度。然后将RNA干扰的Cal27细胞接种于裸鼠颈部皮下左、右两侧,定期监测肿瘤体积,绘制肿瘤生长曲线,称取瘤体重量。采用免疫组织化学检测移植瘤组织中TCAB1、B细胞淋巴瘤蛋白-2(Bcl-2)、半胱氨酸天冬氨酸特异性蛋白酶-3、血管内皮生长因子(VEGF)和细胞周期蛋白D1的表达。结果 Q-FISH结果显示,sh TCAB1处理后的Cal27细胞较sh NC处理后及野生型Cal27细胞的端粒平均长度明显缩短。sh TCAB1处理组OSCC移植瘤生长速度明显较sh NC对照组减慢。移植瘤平均重量为0.06 g,对照组为0.26 g,抑瘤率达76.9%。免疫组织化学结果显示,sh TCAB1组移植瘤中TCAB1表达明显降低,同时,Bcl-2、VEGF和细胞周期蛋白D1的表达降低,半胱氨酸天冬氨酸蛋白酶-3的表达升高。结论 sh TCAB1慢病毒能够明显抑制裸鼠人OSCC移植瘤的生长,其抗肿瘤机制与诱导肿瘤细胞凋亡、延长肿瘤细胞分裂周期及抑制血管生成有关。
[Abstract]:Objective to investigate the effect of telomerase card body protein 1 (TCAB1) gene silencing on the growth and biological behavior of tumor cells in nude mice. Methods the chromosome ends of Cal27 cell line of 26 generations of human luminal squamous cell carcinoma (OSCC) were stably screened by quantitative fluorescence in situ hybridization (Q-FISH) after treatment with RNA interference lentivirus sh TCAB1 and sh negative control (NC). The signal strength of telomere. Then the Cal27 cells interfered with RNA were inoculated on the left and right sides of the neck of nude mice. The tumor volume was monitored regularly and the tumor growth curve was drawn to weigh the weight of the tumor. The expressions of TCAB1,B cell lymphoma protein-2 (Bcl-2), cystine aspartate specific protease-3, vascular endothelial growth factor (VEGF) and cell cycle D1 were detected by immunohistochemistry. Results the results of Q-FISH showed that the average telomere length of Cal27 cells treated with sh TCAB1 was significantly shorter than that treated with sh NC and wild type Cal27 cells, and the growth rate of OSCC transplantation tumors in sh TCAB1 treatment group was significantly slower than that in sh NC control group. The average weight of the transplanted tumor was 0.06g, and that of the control group was 0.26g. The tumor inhibition rate was 76.9%. The results of immunohistochemistry showed that the expression of TCAB1 was significantly decreased, the expression of Bcl-2,VEGF and cell cycle D1 was decreased, and the expression of cystine aspartate protease-3 was increased in sh TCAB1 group. Conclusion sh TCAB1 lentivirus can significantly inhibit the growth of human OSCC transplantation tumor in nude mice, and its anti-tumor mechanism is related to inducing apoptosis of tumor cells, prolonging the division cycle of tumor cells and inhibiting angiogenesis.
【作者单位】：口腔疾病研究国家重点实验室华西口腔医院(四川大学);
【基金】：国家自然科学基金面上项目(81172579) 四川省科技厅科技支撑计划项目(2013SZ0039)~~
【分类号】：R739.8

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