【摘要】 目的构建pEGFP-N1-DCN真核表达载体并转染胃癌细胞SGC-7901,观察其对细胞增殖、周期、迁移、侵袭的影响。方法将细胞分为3组,分别为空白对照组、pEGFP-N1组及pEGFP-N1-DCN组,质粒通过脂质体转染SGC-7901,RT-PCR及Western blot方法检测表皮生长因子受体(EGFR)及TGF-β1的表达,MTT法检测细胞增殖,流式细胞仪检测细胞周期,Transwell检测核心蛋白聚糖(DCN)对细胞迁移及侵袭能力的影响。结果 RT-PCR及Western blot检测结果显示,DCN使EGFR及TGF-β1的表达均降低(P<0.05);MTT法检测发现72 h时空白对照组、pEGFP-N1组及pEGFP-N1-DCN组细胞D(490)值分别为(1.190±0.037)、(1.169±0.052)、(0.572±0.021),DCN使细胞增殖受到明显抑制(P<0.05);流式细胞仪分析细胞周期空白对照组、pEGFP-N1组及pEGFP-N1-DCN组细胞G0～G1的细胞数分别为(46.12±0.63)、(46.07±0.82)、(56.36±0.92),DCN使细胞阻滞在G0～G1(P<0.05);Transwell迁移、侵袭实验显示DCN使细胞迁移和侵袭能力均降低(P<0.05)。结论 DCN通过抑制肿瘤细胞的增殖、迁移、侵袭等起到抗肿瘤作用。更多还原

【Abstract】 Objective To construct an eukaryotic expression vector encoding decorin(DCN)(pEGFP-N1-DCN) and determine its effect on proliferation,cell cycle,migration and invasion of human gastric cancer cell line SGC-7901 after transfection.Methods SGC-7901 cells were divided into 3 groups,control,pEGFP-N1 group and pEGFP-N1-DCN group.SGC-7901 cells were transfected by corresponding plasmids with liposome.The expressions of epidermal growth factor receptor(EGFR) and transforming growth factor-β1(TGF-β1) were detected by Western blotting and RT-PCR,cell proliferation by MTT assay,cell cycle by flow cytometry and cell migration and invasion by Transwell chamber test.Results RT-PCR and Western blotting results showed that the expressions of EGFR and TGF-β1 were decreased in pEGFP-N1-DCN group.MTT assay demonstrated that the value of D(490) of the control group,pEGFP-N1 group and pEGFP-N1-DCN group were 1.190±0.037,1.169±0.052,and 0.572±0.021 respectively,indicating that DCN made cell proliferation inhibited(P<0.05).Flow cytometric analysis showed that cell population at G0 to G1 stage of these 3 groups were(46.12±0.63)%,(46.07±0.82)%,and(56.36±0.92)%,and the cells in pEGFP-N1-DCN group were arrested at G0 to G1 stage(P<0.05).Transwell assay showed that DCN inhibits cell migration and invasion(P<0.05).Conclusion DCN plays a role of anti-tumor by inhibiting tumor cell proliferation,migration and invasion.更多还原