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目的检测SOX15在结肠癌细胞株中表达水平以及对细胞增殖、凋亡、迁移侵袭的影响。方法应用RT-PCR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水平。通过脂质体Lipofectamine 2000将重组质粒p EGFP-N1-SOX15及空质粒p EGFP-N1转染进Caco2细胞,应用RT-PCR及Western blot分别检测SOX15在Caco2细胞中的m RNA以及蛋白表达水平;CCK-8检测SOX15对Caco2细胞活力的影响,克隆形成实验观察细胞增殖变化,Transwell检测SOX15对细胞迁移及侵袭能力的调控,流式细胞仪分析细胞凋亡率。结果 SOX15在Caco2、SW480、HT29、HCT116结肠癌细胞中m RNA及蛋白质表达明显低于其在正常组织中的表达水平(P<0.01)。p EGFP-N1-SOX15转染组细胞m RNA(1.23±0.10)和蛋白(1.13±0.08)表达显著高于p EGFP-N1对照组[(0.54±0.06)、(0.19±0.03),P<0.01]及空白对照组[(0.51±0.03)、(0.14±0.02),P<0.01];与空白对照组及p EGFP-N1对照组相比,p EGFP-N1-SOX15转染组可明显抑制细胞增殖及迁移侵袭能力(P<0.01),并诱导细胞发生凋亡(P<0.01)。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Caco2的增殖及迁移、侵袭能力,并促进细胞凋亡。
Objective To detect the expression of SOX15 in colon cancer cell lines and its effect on cell proliferation, apoptosis, migration and invasion. Methods The expression of SOX15 in Caco2, SW480, HT29, HCT116 colon cancer cells and normal colon tissues was detected by RT-PCR. The recombinant plasmids p EGFP-N1-SOX15 and empty plasmid p EGFP-N1 were transfected into Caco2 cells by Lipofectamine 2000. The mRNA and protein levels of SOX15 in Caco2 cells were detected by RT-PCR and Western blot respectively. The effect of SOX15 on the viability of Caco2 cells was detected by CCK-8. The proliferation of Caco2 cells was detected by clonogenic assay. The effect of SOX15 on cell migration and invasion was analyzed by Transwell assay. The apoptosis rate was analyzed by flow cytometry. Results The mRNA and protein expressions of SOX15 in Caco2, SW480, HT29 and HCT116 colon cancer cells were significantly lower than those in normal tissues (P <0.01). The expression of m RNA (1.23 ± 0.10) and protein (1.13 ± 0.08) in the EGFP-N1-SOX15 group was significantly higher than that in the p EGFP-N1 group [(0.54 ± 0.06), ] And blank control group [(0.51 ± 0.03), (0.14 ± 0.02), P <0.01]. Compared with blank control group and p EGFP-N1 control group, p EGFP-N1-SOX15 transfected group could significantly inhibit the proliferation of cells Proliferation and migration (P <0.01), and induce cell apoptosis (P <0.01). Conclusion SOX15 is overexpressed in colon cancer cells. Overexpression of SOX15 can significantly inhibit the proliferation, migration and invasion of Caco2 cells and promote the apoptosis of colon cancer cells.