目的 探讨NET-1表达与子宫颈癌微血管生成的关系及其分子机制.方法 采用免疫组化MaxVision法检测80例子宫颈鳞癌、10例慢性子宫颈炎组织中NET-1、NF-κB p65、VEGF蛋白表达.采用CD34内皮标记,结合Weinner计数法检测子宫颈鳞癌组织的微血管密度(microvascular density,MVD).通过阳离子脂质体将真核表达质粒pCMV6-NET-1转染子宫颈鳞癌细胞以获得NET-1基因过表达;利用NF-κB特异抑制剂PDTC处理子宫颈鳞癌细胞以抑制NF-κB的激活;应用Western blot法检测细胞蛋白的表达.结果 子宫颈鳞癌组织中NET-1呈高表达,NET-1表达与子宫颈鳞癌MVD、NF-κB p65和VEGF蛋白表达呈正相关.NET-1基因转染引起子宫颈鳞癌SiHa细胞胞核NF-κB p65和胞质VEGF水平上高,应用NF-κB抑制剂PDTC预处理SiHa细胞能显著抑制NET-1引起的VEGF表达上调.结论 NET-1促进子宫颈鳞癌微血管生成,其机制与NET-1激活子宫颈鳞癌NF-κB信号途径上调VEGF表达有关.%Purpose To explore the association of NET-1 expression with the angiogenesis of cervical squamous cell carcinoma and the related molecular mechanism.Methods Immunohistochemistry MaxVision was used to detect the expression of NET-1,NF-κB p65 and VEGF in 80 cases of cervical squamous cancer tissue and 10 cases of chronic cervicitis tissue.The microvascular density (MVD) was assessed by labeling endothelia with CD34-antibody and counted according to Weinner's standard.The eukaryotic expression plasmid pCMV6-NET-1 was transfected into cervical squamous cancer cells by cationic liposome in order to obtain over expression of NET-1 gene.In order to inhibit the activation of NF-kappa B,cervical squamous carcinoma cells were treated with NF-kappa B specific inhibitor PDTC.The protein expression in SiHa cells was assessed by Western blotting.Results The expression level of NET-1 in the cervical squamous carcinoma tissue was significantly higher than that in the tissue of control group.NET-1 expression was positively correlated to the MVD,NF-κB p65 and VEGF expression,respectively.Transient transfection of NET-1 gene significantly increased the nuclear expression of NF-κB p65 and cytoplasm expression of VEGF in the SiHa cells.However,pretreatment of SiHa cells with NF-inhibitor PDTC significantly inhibited NET-1 induced upregulation of VEGF expression.Conclusion NET-1 promotes the angiogenesis of cervical squamous carcinoma through the activation of the NF-κB signaling which upregulate the expression of VEGF in the cancer cells.
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