Skp2及p27kip1在原发性胃癌组织的表达及对胃癌细胞增殖、凋亡的影响

Expression of Skp2 and p27kip1 in primary gastric cancer and their effects on proliferation and apoptosis of gastric cancer cells

目的探究S期激酶相关蛋白2(S-phase kinase associated protein 2,Skp2)及细胞周期素依赖的蛋白激酶抑制物(Cyclin dependent protein kinase inhibitor,p27kip1)在原发性胃癌组织的表达及si-Skp2对胃癌细胞增殖、凋亡和p27Kip1表达的影响。方法组织学实验:临床入组2018年12月~2021年12月唐山市开滦总医院林西医院归档的资料齐全完整的63例原发性胃癌患者,获得患者的癌组织和癌旁组织,通过qPCR对两组患者胃癌组织和胃癌旁组织Skp2和p27Kip1mRNA表达水平进行检测。细胞学实验,将人胃癌细胞SGC-7901分为对照组和si-Skp2组,对照组转染NC质粒,si-Skp2组转染si-Skp2质粒,CCK8法检测各组细胞24 h、48 h和72 h增殖能力,流式细胞分别分析两组细胞细胞存活、早期凋亡和晚期凋亡,Western blot检测两组细胞Skp2和p27kip1蛋白表达水平。结果组织学实验表明,相比于癌旁组织,癌组织中Skp2mRNA表达显著上升,p27Kip1表达显著降低,差异有统计学意义(P<0.01)。细胞学实验表明,与对照组相比,si-Skp2组细胞24 h、48 h和72 h细胞增殖能力显著降低;细胞存活显著降低、细胞晚期凋亡显著增加(P<0.01),细胞早期凋亡没有显著的统计学差异(P>0.05);Western blot实验表明si-Skp2组细胞Skp2表达显著降低,p27Kip1蛋白表达水平显著上调(P<0.01)。结论原发性胃癌组织中Skp2表达显著高表达,p27Kip1显著低表达,si-Skp2具有显著抑制胃癌细胞增殖,促进细胞晚期凋亡的作用,其机制与p27Kip1的调控相关。

Objective To explore S-phase kinase associated protein 2(Skp2)and cyclin dependent protein kinase inhibitors Expression of cyclin dependent protein kinase inhibitor(p27kipl)in primary gastric cancer tissue and its effect on proliferation and apoptosis of gastric cancer cells.Methods Histological experiment:63 patients with primary gastric cancer were enrolled from December 2018 to December 2021 in Linxi Hospital of Kailuan General Hospital of Tangshan city.The cancer tissues and adjacent tissues of the patients were obtained.The expression levels of Skp2 and p27kipl mRNA in gastric cancer tissues and adjacent tissues were detected by qPCR.Cytological experiment:human gastric cancer cell line SGC-7901 was divided into control group and si Skp2 group.NC plasmid was transfected into control group and si-Skp2 plasmid was transfected into si Skp2 group.The proliferation ability of cells in each group was detected by CCK8 method at 24 h,48 h and 72 h.The cell survival,early apoptosis and late apoptosis of the two groups were analyzed by flow cytometry.The expression levels of Skp2 and p27kipl protein were detected by Western blot.Results Histological experiments showed that compared with adjacent tissues,the expression of Skp2 mRNA increased significantly and the expression of p27kipl decreased significantly,(P<0.01).Cytological experiments showed that compared with the control group,the cell proliferation ability of si-Skp2 group decreased significantly at 24 h,48 h and 72 h,cell survival decreased significantly,late apoptosis increased significantly(P<0.01),and there was no significant difference in early apoptosis(P>0.05);Western blot showed that the expression of Skp2 decreased significantly and the expression level of p27kipl Protein increased significantly in si Skp2 group(P<0.01).Conclusion The expression of Skp2 was significantly higher and p27kipl was significantly lower in primary gastric cancer.Si-Skp2 could sig-nificantly inhibit the proliferation of gastric cancer cells and promote late apoptosis,and its mechanism was related to the regulation of p27kipl.