沉默MMP-2对人宫颈鳞癌SiHa细胞生物学行为的影响

Effect of Silencing MMP-2 on Biological Behavior in Human Cervical Squamous SiHa Cells

目的:研究基质金属蛋白酶2(MMP-2)对人宫颈鳞癌SiHa细胞增殖、迁移、侵袭及裸鼠成瘤能力的影响。方法:分别用MMP-2沉默慢病毒感染人宫颈鳞癌SiHa细胞(MMP-2-LV组)、空载慢病毒感染人宫颈鳞癌SiHa细胞为阴性对照组(NC-LV组)。转染后采用实时荧光定量聚合酶链反应(qRT-PCR)观测MMP-2 mRNA的表达,蛋白质印迹(Western blotting)检测MMP-2蛋白的表达,流式细胞技术、Transwell侵袭、迁移和CCK-8细胞增殖实验观测MMP-2对细胞凋亡、侵袭、迁移和细胞增殖的影响,在裸鼠上构建移植瘤模型观测MMP-2对小鼠皮下成瘤能力。结果:转染后,与NC-LV组比较,MMP-2-LV组细胞MMP-2 mRNA和蛋白的表达量较低(P<0.05),增殖能力降低(P<0.01),侵袭、迁移能力降低(P<0.05),而细胞凋亡率升高(P<0.05)。裸鼠成瘤实验显示,接种MMP-2-LV组细胞的裸鼠皮下成瘤体积小于接种NC-LV组细胞的裸鼠皮下成瘤体积(P<0.01)。结论:沉默MMP-2可抑制SiHa细胞增殖、侵袭及迁移能力并促进其凋亡,MMP-2可能是宫颈鳞癌的潜在诊疗靶点。

Objective:To investigate the effect of matrix metalloproteinase 2(MMP-2)on cell proliferation,migration,invasion of human cervical squamous carcinomas SiHa cells and tumorigenic ability of nude mice.Methods:Cervical squamous carcinoma SiHa cells were transfected with MMP-2 silencing lentiviral vector(MMP-2-LV group),and cervical squamous carcinoma SiHa cells were transfected with a control lentiviral vector(NC-LV group,negative control).The mRNA levels of MMP-2 were measured using qRT-PCR.The expression of MMP-2 protein was detected by Western blotting.The effects of MMP-2 on cell proliferation,invasion,migration and apoptosis were observed by CCK-8 cell proliferation assay,Transwell invasion,migration assay,and flow cytometry.The effect of MMP-2 on subcutaneous tumor formation ability of nude mice was observed by transplanted tumor model in nude mice.Results:Compared with the NC group,the MMP-2-LV group showed lower expression of MMP-2 mRNA and protein(P<0.01),and it showed a reduction of the cell proliferation ability(P<0.01),invasion and migration ability(P<0.05),as well as the increase of apoptosis(P<0.05).The nude mice tumor formation showed lower subcutaneous tumorigenic average tumor volume of nude mice inoculated with MMP-2-LV group than the NC-LV group(P<0.01).Conclusions:Silencing MMP-2 can inhibit the proliferation,invasion and migration of SiHa cells and promote apoptosis,suggesting that MMP-2 could be a potential therapeutic target for cervical squamous cell carcinoma.