秦艽多糖通过调控STMN1表达影响肺癌细胞增殖、迁移及侵袭

Effects of Gentiana Macrophylla Pall Polysaccharide on the Proliferation,Migration and Invasion of Lung Cancer Cells by Regulating STMN1 Expression

目的:探讨秦艽多糖是否可通过调控微管不稳定性蛋白1(STMN1)表达而抑制肺癌细胞增殖、迁移及侵袭。方法:体外培养肺癌细胞A549,随机分为对照组、秦艽多糖低、中、高浓度组(25,50,100μg·ml^(-1));采用甲基噻唑基四唑(MTT)与平板克隆形成实验检测细胞增殖能力;Transwell小室实验检测细胞迁移及侵袭;pcDNA 3.1-STMN1转染至A549细胞,采用上述方法检测细胞增殖、迁移及侵袭;蛋白免疫印迹法(Western blot)检测STMN1、增殖标记蛋白细胞增殖核抗原-67(Ki67)、神经型钙黏蛋白(N-cadherin)、上皮型钙黏蛋白(E-cadherin)的表达量。结果:与对照组比较,秦艽多糖中、高浓度组细胞活力、克隆形成数、迁移细胞数、侵袭细胞数及STMN1、Ki67、N-cadherin蛋白水平显著降低(P<0.05),E-cadherin蛋白水平显著升高(P<0.05),且秦艽多糖各浓度组间的指标差异均有统计学意义(P<0.05);与高浓度秦艽多糖+pcDNA 3.1组比较,高浓度秦艽多糖+pcDNA 3.1-STMN1组细胞活力、克隆形成数、迁移细胞数、侵袭细胞数及STMN1、Ki67、N-cadherin蛋白水平显著升高(P<0.05),E-cadherin蛋白水平显著降低(P<0.05)。结论:秦艽多糖可能通过下调STMN1的表达从而抑制肺癌细胞增殖、迁移及侵袭。

Objective:To investigate the inhibitory effects of gentiana macrophylla pall polysaccharide on the proliferation,migration and invasion of lung cancer cells by regulating the expression of stathmin 1(STMN1).Methods:Lung cancer cells A549 were cultured in vitro and randomly divided into control group and gentiana macrophylla pall polysaccharide low,medium and high-dose groups(25,50,100μg·ml^(-1)).MTT and plate clone formation experiments were used to detect cell proliferation ability.Transwell cell test was used to detect cell migration and invasion.pcDNA 3.1-STMN1 was transfected into A549 cells,and cell proliferation,migration and invasion were detected by the above methods.Western blot was used to detect the expression levels of STMN1,Ki67,N-cadherin and E-cadherin.Results:Compared with those in the control group,the cell viability,number of clones formed,number of migrating cells,number of invasive cells and protein levels of STMN1,Ki67 and N-cadherin in gentiana macrophylla pall polysaccharide medium and high-dose groups significantly reduced(P<0.05),the protein levels of E-cadherin were significantly increased(P<0.05),and there were statistically significant differences in various indexes among gentiana macrophylla pall polysaccharide low,medium and highdose groups(P<0.05).Compared with those in gentiana macrophylla pall polysaccharide high-dose+pcDNA 3.1 group,the cell viability,number of clones formed,number of migrating cells,number of invasive cells and protein levels of STMN1,Ki67 and N-cadherin in gentiana macrophylla pall polysaccharide high-dose+pcDNA 3.1-STMN1 group were significantly increased(P<0.05),and the protein levels of E-cadherin were significantly decreased(P<0.05).Conclusion:Gentiana macrophylla pall polysaccharide may inhibit the proliferation,migration and invasion of lung cancer cells by down-regulating the expression of STMN1.