Nck1蛋白通过Rac1-PAK1-MMP2信号通路对宫颈鳞癌组织血管生成的调控机制研究

Study on the regulatory mechanism of Nck1 protein on angiogenesis in cervical squamous cell based on Rac1-PAK1-MMP2 signaling pathway.

目的Nck1蛋白通过Rac1-PAK1-MMP2信号通路对宫颈鳞癌组织血管生成的调控机制研究。方法回顾性选择2018年10月至2019年10月福建中医药大学附属人民医院的宫颈鳞癌病理组织标本80例,根据癌变组织是否发生侵袭分为侵袭组(n=38)与未侵袭组(n=42),另选择子宫颈鳞状上皮正常组织作为对照组(n=30)。采用免疫组织化学法检测3组标本中Nck1蛋白表达水平,利用免疫组织化学法观察侵袭组与非侵袭组标本中微血管密度(MVD)。通过制备Nck1过表达、低表达的宫颈癌细胞siHa,人脐静脉内皮细胞ECV304,测定宫颈癌细胞中Rac1、PAK1、MMP2表达水平,经Nck1过表达、低表达的siHa上清液干扰后的ECV304细胞增殖能力、血管形成能力、迁移能力。结果侵袭组Nck1蛋白表达水平明显高于非侵袭组与对照组,非侵袭组Nck1蛋白表达水平高于对照组,差异均有统计学意义(P<0.05)。侵袭组MVD与肿瘤直径明显大于非侵袭组,差异均有统计学意义(P<0.05)。宫颈鳞癌患者Nck1蛋白表达水平与MVD呈正相关关系(P<0.05)。Nck1过表达的siHa细胞中Rac1、PAK1、MMP2表达水平均明显高于Nck1低表达的siHa细胞,差异均有统计学意义(P<0.05)。经Nck1过表达的siHa细胞上清液干扰后的ECV304增殖率、迁移率、管腔形成数均高于经Nck1低表达的siHa细胞上清液干扰后的ECV304细胞,差异均有统计学意义(P<0.05)。结论宫颈病理标本中Nck1蛋白处于高表达状态,其中已发生癌细胞侵袭、迁移的标本Nck1蛋白表达水平更高,过表达的Nck1能够促进血管内皮细胞增殖、迁移,这一作用机制与宫颈癌细胞中Rac1/PAK1/MMP2信号通路有关。

Objective To investigate the regulatory mechanism of Nck1 protein on angiogenesis in cervical squamous cell Based on Rac1-PAK1-MMP2 signaling pathway.Methods From October 2018 to October 2019,80 cases of cervical squamous cell carcinoma pathological tissue specimens from the Affiliated People's Hospital of Fujian University of Traditional Chinese Medicine were retrospectively selected.They were divided into the invasion group(n=38)and the non-invasion group(n=42)according to whether the cancerous tissue was invaded,and the normal cervical squamous epithelial tissue was selected as the control group(n=30).Immunohistochemistry was used to detect the expression of Nck1 protein in the three groups,and the microvessel density(MVD)in the invasion group and non-invasive group.The expression levels of Rac1,PAK1 and MMP2 in siHa with overexpression and underexpression of Nck1 were determined,and the proliferation,angiogenesis and migration ability of ECV304 cells transfected with siHa supernatant with overexpression and underexpression of Nck1 were determined.Results The protein expression level of Nck1 among three groups was the highest in the invasion group,followed by non-invasion group and control group,the differences were statistically significant(P<0.05).The MVD and tumor diameters in the invasion group were significantly larger than those in the non-invasive group,and the differences were statistically significant(P<0.05).Nck1 protein expression level was positively correlated with MVD in patients with cervical squamous cell carcinoma,the differences were statistically significant(P<0.05).The expression levels of Rac1,PAK1 and MMP2 in siHa cells with overexpressed Nck1 were significantly higher than those in siHa cells with underexpressed Nck1,the differences were statistically significant(P<0.05).The proliferation rate,migration rate and lumen formation number of ECV304 transfected with siHa supernatant with overexpressed Nck1 were higher than those of ECV304 cells transfected with siHa supernatant with underexpressed Nck1,the differences were statistically significant(P<0.05).Conclusion Nck1 protein is highly expressed in cervical pathological specimens,among which the expression level of Nck1 protein is higher in the samples with cell invasion and migration.Overexpressed Nck1 can promote proliferation and migration of vascular endothelial cells,which is related to the Rac1/PAK1/MMP2 signaling pathway in cervical cancer cells.