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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。
Objective To construct pEGFP-N1-DCN eukaryotic expression vector and transfect it into SGC-7901 gastric cancer cell line SGC-7901 to observe its effect on cell proliferation, cycle, migration and invasion. Methods The cells were divided into three groups: blank control group, pEGFP-N1 group and pEGFP-N1-DCN group. The plasmids were transfected into SGC-7901 by lipofectamine. The expression of epidermal growth factor receptor (EGFR) and TGF-β1 were detected by MTT assay. Cell proliferation was detected by MTT assay. Cell cycle was detected by flow cytometry. Transwell assay was used to detect the effect of decorin on cell migration and invasion. Results The results of RT-PCR and Western blot showed that the expression of EGFR and TGF-β1 were decreased in DCN group (P <0.05). The MTT assay showed that the expression of EGFR and TGF-β1 in the blank control group, pEGFP-N1 group and pEGFP- Cell D (490) values were (1.190 ± 0.037), (1.169 ± 0.052) and (0.572 ± 0.021), respectively, which were significantly inhibited by DCN (P <0.05). Flow cytometry analysis of cell cycle blank control group, The number of G0 ~ G1 cells in pEGFP-N1 group and pEGFP-N1-DCN group were (46.12 ± 0.63), (46.07 ± 0.82) and (56.36 ± 0.92), respectively. DCN blocked cells in G0 ~ G1 (P < 0.05). Transwell migration and invasion assays showed that DCN reduced the ability of cell migration and invasion (P <0.05). Conclusion DCN can inhibit the proliferation, migration and invasion of tumor cells.