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目的探讨 PTEN 基因对胃癌细胞生物学行为的影响及其机制。方法脂质体转染并经免疫细胞化学、Western blot 鉴定得到稳定高表达 PTEN 蛋白的 SGC7901细胞克隆;细胞倍增时间测定、平板克隆形成试验及流式细胞术分析转染前后细胞增殖能力及细胞周期与凋亡率的变化;应用 ELISA 法、明胶酶谱分析、免疫细胞化学及 Western blot 等技术分别检测转染前后细胞中血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-9与 MMP-2蛋白的分泌与表达。结果成功地建立了稳定高表达 PTEN 蛋白的 SGC7901细胞模型。PTEN-SGC7901细胞(PTEN 基因转染细胞组)的倍增时间较 SGC7901(未转染细胞组)、PBP-SGC7901细胞(空白质粒转染细胞组)显著延长(P<0.05)。PTEN-SGC7901细胞的克隆形成率明显低于对照组,PTEN-SGC7901细胞对 SGC7901、PBP-SGC7901细胞的集落抑制率分别为69.8%、64.8%。PTEN-SGC7901组细胞的 G_1期细胞含量明显较对照组增多(P<0.05),但三组细胞之间的凋亡率无明显差别(P>0.05)。PTEN-SGC7901细胞 VEGF 与MMP-9蛋白的表达和分泌明显低于对照组(P<0.05),但 MMP-2蛋白在三组间的表达无差异(P>0.05)。结论 PTEN 可抑制胃癌细胞株 SGC7901细胞的生长与增殖;PTEN 可能通过下调 SGC7901细胞株 VEGF 及 MMP-9的表达与分泌来抑制肿瘤浸润与转移的发生。
Objective To investigate the effect of PTEN gene on the biological behavior of gastric cancer cells and its mechanism. Methods Liposomes were transfected into SGC7901 cells, which were stably expressed PTEN protein by immunocytochemistry and Western blot. The cell doubling time, plate clone formation assay and flow cytometry were used to analyze the cell proliferation and cell cycle And apoptosis rate. The levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP) -9 and MMP-9 were detected by ELISA, gelatin zymography, immunocytochemistry and Western blot before and after transfection. 2 protein secretion and expression. Results The SGC7901 cell model was successfully established to stably express PTEN protein. The doubling time of PTEN-SGC7901 cells (PTEN gene transfected cells) was significantly longer than that of SGC7901 (untransfected cells) and PBP-SGC7901 cells (blank transfected cells) (P <0.05). The colony formation rate of PTEN-SGC7901 cells was significantly lower than that of the control group. The colony-inhibitory rates of PTEN-SGC7901 cells to SGC7901 and PBP-SGC7901 cells were 69.8% and 64.8%, respectively. The cell number of G_1 phase in PTEN-SGC7901 group was significantly higher than that in control group (P <0.05), but there was no significant difference between the three groups (P> 0.05). The expression and secretion of VEGF and MMP-9 in PTEN-SGC7901 cells were significantly lower than those in the control group (P <0.05). However, there was no difference in the expression of MMP-2 between the three groups (P> 0.05). Conclusion PTEN can inhibit the growth and proliferation of gastric cancer cell line SGC7901. PTEN may inhibit the invasion and metastasis of tumor by down-regulating the expression and secretion of VEGF and MMP-9 in SGC7901 cell line.