Jagged1对非小细胞肺癌细胞增殖、转移和凋亡的影响及机制研究

Effects of Jagged1 on proliferation, metastasis and apoptosis of non small cell lung cancer cells and its action mechanism

目的探讨Jagged1对非小细胞肺癌细胞增殖、转移和凋亡的影响及作用机制。方法采用Western blot检测Jagged1在正常支气管上皮细胞系16-HBE和非小细胞肺癌细胞系NCI-H1975、A549、NCI-H520、SK-MES-1中的表达情况。将体外培养的A549细胞分为对照组(正常培养)、NC-siRNA组(转染阴性对照)和Jagged1-siRNA组(转染Jagged1干扰序列Jagged1-siRNA),采用Western blot检测转染效果及STAT3信号通路相关蛋白p-STAT3、STAT3、CyclinD1、Bcl-2、Survivin、VEGF的表达情况,MTT法、克隆形成实验、Transwell小室、划痕实验和流式细胞仪分别检测细胞增殖、克隆形成、侵袭、迁移和凋亡能力。结果NCI-H1975、A549、NCI-H520、SK-MES-1细胞中Jagged1蛋白的表达水平较16-HBE细胞均明显升高(P<0.05),且A549细胞中最高,SK-MES-1细胞中最低。与对照组比较,Jagged1-siRNA组细胞中Jagged1、CyclinD1、Bcl-2、Survivin、VEGF蛋白的表达水平和STAT3磷酸化水平均明显降低,同时细胞增殖、克隆形成、侵袭和迁移能力均明显减弱,而细胞凋亡能力明显增强(P<0.05);而NC-siRNA组和对照组比较未见明显改变(P>0.05)。结论Jagged1在非小细胞肺癌细胞中高表达,干扰Jagged1表达可抑制非小细胞肺癌细胞增殖、侵袭、迁移并促进细胞凋亡,其作用机制可能与抑制STAT3信号通路的活化有关。

Objective To investigate the effect of Jagged1 on proliferation,metastasis and apoptosis of non small cell lung cancer(NSCLC)cells,and to explore its action mechanism.Methods The expression levels of Jagged1 in normal bronchial epithelial cell line 16HBE and non small cell lung cancer cell lines NCI-H1975,A549,NCI-H520 and SK-MES-1 were detected by Western Blot.The A549 cells cultured in vitro were divided into control group(normal culture),NC-siRNA group(transfection negative control)and Jagged1-siRNA group(transfected with Jagged1 interference sequence Jagged1 siRNA),the transfection effects and the expression levels of STAT3 signal pathway related proteins including pSTAT3,STAT3,CyclinD1,Bcl2,Survivin and VEGF were detected by Western Blot.Moreover the cell proliferation,colony formation,invasion,migration and apoptosis capacity were detected respectively by MTT assay,colony formation assay,transwell chamber,scratch assay and flow cytometry.Results The expression levels of Jagged1 protein in NCI-H1975,A549,NCI-H520 and SK-MES-1 cells were significantly higher than those in 16 HBE cells(P<0.05),and the expression levels of Jagged1 protein were the highest in A549 cells and the lowest in SK-MES-1 cells.As compared with those in control group,the expression levels of Jagged1,Cyclin D1,Bcl2,Survivin,VEGF protein and STAT3 phosphorylation in Jagged1 siRNA group were significantly decreased,meanwhile the cell proliferation,cloning,invasion and migration were significantly weakened,however the cell apoptosis ability was significantly enhanced(P<0.05).There were no significant differences in above indexes between NC-siRNA group and control group(P>0.05).Conclusion Jagged1 is highly expressed in NSCLC cells.To interference with Jagged1 expression can inhibit the proliferation,invasion,migration and promote apoptosis of NSCLC cells,and its action mechanism may be related to the inhibition of STAT3 signaling pathway activation.