铜绿假单胞菌注射液对NCI-N87胃癌细胞株增殖能力的影响及其机制研究

Effect of Pseudomonas Aeruginosa Mannose-sensitive Hemagglutinin on Proliferation of Gastric Cancer NCI-N87 Cells and Its Mechanism

目的探讨铜绿假单胞菌注射液(Pseudomonas aeruginosa mannose-sensitive hemagglutinin,PA-MSHA)对NCI-N87胃癌细胞增殖能力的影响及其分子作用机制。方法MTT法检测PA-MSHA对NCI-N87胃癌细胞增殖的影响,流式细胞术检测细胞凋亡率,划痕实验和侵袭实验检测细胞迁移和侵袭能力的变化,WesternBlot检测PTEN蛋白和p-AKT蛋白的表达变化,转染PTEN siRNA敲低PTEN表达后,再次行MTT法检测PA-MSHA对细胞增殖的影响。结果PA-MSHA抑制NCI-N87胃癌细胞的增殖具有剂量和时间依赖性。与对照组比较,NCI-N87细胞经PA-MSHA处理后,凋亡率显著增加(P<0.05),迁移和侵袭能力显著下降(P<0.05)。与对照组比较,PA-MSHA显著提高PTEN蛋白表达,降低p-AKT蛋白表达(P<0.05)。通过siRNA转染敲低PTEN表达后,PA-MSHA抑制NCI-N87细胞增殖的能力较对照组显著下降(P<0.05)。结论PA-MSHA对NCI-N87胃癌细胞具有抑制增殖、促进凋亡、抑制侵袭转移的作用,其机制与PTEN/AKT信号通路有关。

OBJECTIVE To investigate the effect of Pseudomonas aeruginosa mannose-sensitive hemagglutinin(PA-MSHA)on the proliferation of gastric cancer NCI-N87 cells and its mechanism.METHODS MTT assay was used to detect the effect of PA-MSHA on the proliferation of gastric cancer cells NCI-N87.The apoptosis rate was tested by flow cytometry.The ability of cell migration and invasion was detected by wound healing assay and invasion assay.Western Blot was carried out to examine the protein expression of PTEN and p-AKT.MTT assay was used to further examine the effect of PA-MSHA on cell proliferation after PTEN siRNA transfection.RESULTS PA-MSHA inhibited the proliferation of gastric cancer cells NCI-N87 in a dose-and time-dependent manner.Compared with control group,after treatment with PA-MSHA,the apoptosis rate of NCI-N87 cells was significantly increased(P<0.05),and the migration and invasion ability of NCI-N87 cells was significantly decreased(P<0.05).Compared with control group,PA-MSHA significantly increased the expression of PTEN protein and decreased the level of p-AKT protein(P<0.05).Knock down of PTEN by siRNA transfection weakened the ability of PA-MSHA on inhibiting the proliferation of NCI-N87 cells compared with control group(P<0.05).CONCLUSION PA-MSHA can inhibit proliferation,promote apoptosis and inhibit invasion and metastasis of gastric cancer cell line NCI-N87,and its mechanism may be related to PTEN/AKT signaling pathway.