铜绿假单胞菌注射液对肺癌A549细胞自噬及PI3K/Akt通路的影响

目的探究铜绿假单胞菌注射液(PA-MSHA)对肺癌A549细胞自噬及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法体外培养人肺癌A549细胞,随机分为空白对照组、LY294002组(加入20μmol/L LY294002),PA-MSHA干预组(加入0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA)。干预培养48 h后,MTT法检测各组细胞活性,平板克隆形成实验检测各组细胞增殖能力,透射电子显微镜观察各组细胞自噬情况,Western blot检测各组细胞中自噬相关蛋白p62、LC3Ⅰ、LC3Ⅱ及PI3K/Akt通路相关蛋白PI3K、Akt、p-PI3K、p-Akt表达。结果与空白对照组相比,LY294002组、0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05);与LY294002组相比,0.5×10^9/mL、1.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著降低(P<0.05),自噬体个数减少,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著升高(P<0.05);与0.5×10^9/mL、1.0×10^9/mL PA-MSHA组相比,2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05)。结论PA-MSHA可能通过调控PI3K/Akt信号通路,促进肺癌A549细胞自噬,抑制细胞增殖。

Effect of Pseudomonas aeruginosa injection on autophagy and PI3K/Akt pathway of lung cancer A549 cells

Objective To investigate the effect of Pseudomonas aeruginosa injection(PA-MSHA)on autophagy and phosphatidylinositol-3 kinase(PI3K)/protein kinase B(Akt)pathway in lung cancer A549 cells.Methods Human lung cancer A549 cells were cultured in vitro and randomly divided into the blank control group,the LY294002 group(added 20μmol/L LY294002),and the PA-MSHA intervention group(added 0.5×10^9/mL,1.0×10^9/mL,2.0×10^9/mL PA-MSHA).After 48 hours of culture intervention,MTT method was used to detect the cell activity of each group,the experiment of plate clone formation was used to detect the cell proliferation ability of each group,transmission electron microscope was used to observe the autophagy,and Western blot was used to detect the expressions of autophagy related proteins p62,LC3Ⅰ,LC3Ⅱand PI3K/Akt pathway related proteins PI3K,Akt,p-PI3K and p-Akt.Results Compared with the blank control group,the inhibition rate of cell proliferation and the expression of LC3Ⅱ/LC3Ⅰprotein in the LY294002 group,and the 0.5×10^9/mL,1.0×10^9/mL and 2.0×10^9/mL PA-MSHA groups were significantly higher(P<0.05),the number of autophagy were higher,and the rate of cell clone formation,the expressions of p62,p-PI3K/PI3K,p-Akt/Akt proteins were significantly lower(P<0.05).Compared with the LY294002 group,the inhibition rate of cell proliferation and the expression of LC3Ⅱ/LC3Ⅰprotein in the 0.5×10^9/mL and 1.0×10^9/mL PA-MSHA groups were significantly lower(P<0.05),the number of autophagy were lower,and the rate of cell clone formation,the expressions of p62,p-PI3K/PI3K,p-Akt/Akt proteins were significantly higher(P<0.05).Compared with the 0.5×10^9/mL and 1.0×10^9/mL PA-MSHA groups,the inhibition rate of cell proliferation and the expression of LC3Ⅱ/LC3Ⅰprotein in the 2.0×10^9/mL PA-MSHA group were significantly higher(P<0.05),the number of autophagy were higher,and the rate of cell clone formation,the expressions of p62,p-PI3K/PI3K,p-Akt/Akt proteins were significantly lower(P<0.05).Conclusion PA-MSHA may promote autophagy and inhibit cell proliferation of lung cancer A549 cells by regulating PI3K/Akt signaling pathway.