SOX15基因对结肠癌细胞生物学功能的影响
发布时间：2018-08-28 10:03
【摘要】：目的 检测SOX15基因于结肠癌中的表达高低以及对细胞增殖、凋亡、迁移、侵袭的影响。方法 RT-PCR被用于检测SOX15在Caco2、SW480、HT29、HCT116结肠癌细胞以及正常结肠组织中的表达程度高低。通过脂质体Lipofectamine 2000将重组质粒pEGFP-N1-SOX15与空质粒pEGFP-N1转染入Caco2及SW480结肠癌细胞后,应用RT-PCR及Western blot分别检测SOX15在Caco2及SW480细胞中mRNA以及蛋白质表达水平;CCK-8检测SOX15对Caco2及SW480细胞增殖的影响、克隆形成观察癌细胞克隆形成率、流式细胞仪分析细胞凋亡率、Transwell检测SOX15对细胞迁移、侵袭功能的调节控制。结果 与人正常结肠组织相比,SOX15在Caco2、SW480、HT29、HCT116结肠癌细胞中mRNA及蛋白质表达水平明显低于其在正常结肠组织的表达水平。p EGFP-N1-SOX15转染组细胞mRNA和蛋白表达显著高于p EGFP-N1对照组细胞。与pEGFP-N1对照组相比,pEGFP-N1-SOX15转染组可明显抑制细胞增殖及迁移侵袭能力,抑制癌细胞克隆形成率,并诱导细胞凋亡。结论 SOX15在结肠癌细胞中低表达,过表达SOX15可明显抑制结肠癌细胞的增殖及迁移、侵袭能力,并促进细胞凋亡。
[Abstract]:Objective to detect the expression of SOX15 gene in colon cancer and its effect on cell proliferation, apoptosis, migration and invasion. Methods RT-PCR was used to detect the expression of SOX15 in Caco2,SW480,HT29,HCT116 colon cancer cells and normal colon tissues. The recombinant plasmid pEGFP-N1-SOX15 and empty plasmid pEGFP-N1 were transfected into Caco2 and SW480 colon cancer cells by liposome Lipofectamine 2000. The mRNA and protein expression levels of SOX15 in Caco2 and SW480 cells were detected by RT-PCR and Western blot respectively. The effect of SOX15 on the proliferation of Caco2 and SW480 cells was detected by CCK-8. Clone formation was observed and apoptosis rate was analyzed by flow cytometry. Transwell was used to detect the regulation of cell migration and invasion by SOX15. Results compared with normal colon tissue, the expression of mRNA and protein in Caco2,SW480,HT29,HCT116 colon cancer cells was significantly lower than that in normal colon tissues. The expression of mRNA and protein in the transfected cells of p EGFP-N1-SOX15 was significantly higher than that in the control cells of p EGFP-N1. Compared with pEGFP-N1 control group, pEGFP-N1-SOX15 transfection group could significantly inhibit cell proliferation, migration and invasion, inhibit cancer cell clone formation, and induce apoptosis. Conclusion the expression of SOX15 in colon cancer cells is low. Overexpression of SOX15 can significantly inhibit the proliferation, migration, invasion and apoptosis of colon cancer cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35
本文编号：2209062
[Abstract]:Objective to detect the expression of SOX15 gene in colon cancer and its effect on cell proliferation, apoptosis, migration and invasion. Methods RT-PCR was used to detect the expression of SOX15 in Caco2,SW480,HT29,HCT116 colon cancer cells and normal colon tissues. The recombinant plasmid pEGFP-N1-SOX15 and empty plasmid pEGFP-N1 were transfected into Caco2 and SW480 colon cancer cells by liposome Lipofectamine 2000. The mRNA and protein expression levels of SOX15 in Caco2 and SW480 cells were detected by RT-PCR and Western blot respectively. The effect of SOX15 on the proliferation of Caco2 and SW480 cells was detected by CCK-8. Clone formation was observed and apoptosis rate was analyzed by flow cytometry. Transwell was used to detect the regulation of cell migration and invasion by SOX15. Results compared with normal colon tissue, the expression of mRNA and protein in Caco2,SW480,HT29,HCT116 colon cancer cells was significantly lower than that in normal colon tissues. The expression of mRNA and protein in the transfected cells of p EGFP-N1-SOX15 was significantly higher than that in the control cells of p EGFP-N1. Compared with pEGFP-N1 control group, pEGFP-N1-SOX15 transfection group could significantly inhibit cell proliferation, migration and invasion, inhibit cancer cell clone formation, and induce apoptosis. Conclusion the expression of SOX15 in colon cancer cells is low. Overexpression of SOX15 can significantly inhibit the proliferation, migration, invasion and apoptosis of colon cancer cells.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.35
【参考文献】
相关期刊论文 前1条
1 李雄武;张徽;柳满然;;Sox2下调表达对MDA-MB-231乳腺癌细胞β-catenin与转录中介因子γ的影响[J];第三军医大学学报;2016年01期
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