缺氧诱导因子-1α在皮肤鳞状细胞癌中的表达及促进细胞增殖作用

发布时间：2018-03-21 11:19

本文选题：缺氧诱导因子-1α　切入点：血管内皮生长因子　出处：《华中科技大学》2011年硕士论文　论文类型：学位论文

【摘要】：目的:研究缺氧诱导因子-1α(HIF-1α)、血管内皮生长因子(VEGF)在鳞状细胞癌细胞中的表达情况,及基因干扰HIF-1α后对鳞状细胞癌细胞的影响。方法:应用逆转录PCR( RT-PCR)、蛋白免疫印记(Western Blot)技术比较鳞状细胞癌细胞系(A431细胞)和永生化角质细胞系(hacat细胞)mRNA和蛋白水平表达;及基因敲除HIF-1α基因后HIF-1α、VEGF mRNA和蛋白表达情况;应用流式细胞仪技术和CCK8细胞增殖检测法检测基因敲除HIF-1α基因后A431细胞凋亡和增殖情况。结果:RT-PCR技术检测A431细胞与hacat细胞相比HIF-1α、VEGF mRNA表达增加(P 0.05),敲除HIF-1α基因的A431细胞与未敲除HIF-1α基因的A431细胞相比HIF-1α、VEGF mRNA表达降低(P 0.05);Western Blot技术检测A431细胞与hacat细胞相比蛋白表达增加(P 0.05),敲除HIF-1α基因的A431细胞与未敲除HIF-1α基因的A431细胞相比HIF-1α、VEGF蛋白表达降低(P 0.05);流式细胞仪技术检测敲除HIF-1α基因的A431细胞与未敲除HIF-1α基因的A431细胞相比,凋亡细胞数增加、凋亡细胞比例增高且差异性有意义(P 0.05);CCK8细胞增殖检测法检测敲除HIF-1α基因的A431细胞与未敲除HIF-1α基因的A431细胞相比,细胞增殖速度慢,存在差异性表达(P 0.05)。结论:HIF-1α、VEGF在鳞状细胞癌细胞系A431中的表达明显高于永生化的角质细胞系hacat细胞中的表达。敲除HIF-1α基因后,A431细胞中HIF-1α、VEGF的表达均下降,且凋亡细胞数增加,增殖速度降低。这些均可说明基因敲出HIF-1α后,可抑制VEGF的表达和功能,抑制鳞癌细胞的增殖并促进其凋亡,表明HIF-1α基因靶向治疗可做为治疗鳞状细胞癌的新型手段之一。
[Abstract]:Aim: to study the expression of hypoxia inducible factor-1 伪 (HIF-1 伪) and vascular endothelial growth factor (VEGF) in squamous cell carcinoma (SCC) and the effect of gene interference with HIF-1 伪 on squamous cell carcinoma (SCC). Methods: reverse transcriptase PCR (RT-PCR) technique was used to compare the expression of HIF-1 mRNA and protein in the squamous cell carcinoma cell line (Con A431 cell line) and immortalized keratinocyte cell line (Hhacat cell line), and the expression of HIF-1 伪 mRNA and protein after gene knockout (HIF-1 伪 gene knockout). Apoptosis and proliferation of A431 cells after HIF-1 伪 gene knockout were detected by flow cytometry and CCK8 cell proliferation assay. Results the expression of HIF-1 伪 -VEGF mRNA in A431 cells was higher than that in hacat cells. The expression of HIF-1 伪 -VEGF mRNA in A431 cells with and without HIF-1 伪 gene knockout was lower than that in A431 cells without HIF-1 伪 gene knockout. Western Blot technique was used to detect the expression of HIF-1 伪 -VEGF mRNA in A431 cells compared with hacat cells. The expression of HIF-1 伪 in A431 cells with HIF-1 伪 knockout was significantly lower than that in A431 cells without HIF-1 伪 gene knockout, and the expression of HIF-1 伪 -VEGF protein in A431 cells with HIF-1 伪 knockout gene was detected by flow cytometry compared with A431 cells without HIF-1 伪 gene knockout, and the expression of HIF-1 伪 protein in A431 cells with HIF-1 伪 gene knockout was significantly lower than that of A431 cells without HIF-1 伪 gene knockout by flow cytometry. When the number of apoptotic cells increased, the proportion of apoptotic cells increased and the difference was significant. The proliferation of A431 cells with HIF-1 伪 knockout was slower than that of A431 cells without HIF-1 伪 gene knockout by P0.05CCK8 cell proliferation assay. Conclusion the expression of HIF-1 伪 -VEGF in squamous cell carcinoma cell line A431 was significantly higher than that in immortalized keratinocyte cell line A431. The expression of HIF-1 伪 -VEGF was decreased and the number of apoptotic cells increased after knockout of HIF-1 伪 gene. These results suggest that HIF-1 伪 gene knockout can inhibit the expression and function of VEGF, inhibit the proliferation and promote the apoptosis of squamous cell carcinoma cells, suggesting that the targeted therapy of HIF-1 伪 gene can be used as one of the new methods for the treatment of squamous cell carcinoma.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R739.5

【相似文献】