【作者】 徐娜； 张芳；

【Author】 Na Xu;Fang Zhang;Shanxi Medical University School and Hospital of Stomatology;

【机构】 山西医科大学口腔医学院口腔医院；

【摘要】 目的:探究核因子κB激酶抑制剂(IKKα)调控的乳腺丝氨酸蛋白酶抑制剂(MASPIN)与口腔潜在恶性病变(OPMDs)的关系,并进一步探究MASPIN在OPMDs癌变中的作用。方法:免疫组化和ELISA分别检测MASPIN、IKKα和炎性因子TNF-α、IL-6在OLP、OLK、OSCC及健康对照(HC)组织中的表达和亚细胞定位情况;RT-PCR、Western Blot、ELISA、小干扰RNA IKKα和过表达质粒MASPIN转染实验检测LPS刺激的HOK细胞中MASPIN与IKKα的调节关系及炎症因子TNF-α和IL-6的表达情况。结果:免疫组化结果显示MASPIN在HC、OLP、OLK和OSCC组中表达逐渐下调,而与HC组相比,IKKα在OLK和OSCC组中高表达,并且部分表达于胞核;而且,ELISA结果证明,与OSCC相比,TNF-α和IL-6在OLP和OLK明显低表达(P<0.01),而与HC组相比,TNF-α和IL-6在OLP和OLK组中高表达(P<0.05)。在细胞层面,LPS(10ug/ml)刺激HOK细胞成功模拟了人口腔炎性环境,RT-PCR、Western Blot和ELISA结果证明,IKKα沉默可促进MASPIN表达,并抑制炎性因子的分泌;而MASPIN过表达则抑制了IKKα的表达,同时抑制了炎性因子的分泌。结论:以上结果表明MASPIN可能通过NF-κB非经典途径抑制炎性因子的分泌,而且MASPIN高表达有望成为阻止OPMDs癌变的潜在抑制剂。更多还原

【Abstract】 Objective: To explore the relationship between nuclear factor kappa B kinase inhibitor(IKKα)-mediated mammary serine protease inhibitor(MASPIN) and oral potential malignant disorders(OPMDs) and further explore the role of MASPIN in the carcinogenesis of OPMDs. Methods: Immunohistochemistry staining and ELISA assays were used to detect the expression and subcellular localization of MASPIN, IKKα and inflammatory factors TNF-α, IL-6 in OLP, OLK, OSCC and healthy control(HC) tissues, respectively; RT-PCR, Western Blot, ELISA, siRNA IKKα and overexpression plasmid MASPIN transfection assays were performed in LPS-induced HOK cells to detect the regulated relationship between MASIN and IKKα and the expression of TNF-α and IL-6. Results: Immunohistochemistry showed that the expression of MASPIN was down-regulated from HC to OSCC groups gradually, while the expression of IKKα which partially entered into nucleus was up-regulated in OLK and OSCC group compared with HC group. Moreover, the expression of TNF-α and IL-6 were decreased significantly in OLP and OLK tissues compared with OSCC tissues(P<0.01) but overexpressed in OLP and OLK tissues compared with HC tissues(P<0.05) by ELISA. In a cultured cell model, HOK cells stimulated by LPS(10ug/ml) simulated the inflammatory environment of oral successfully. RT-PCR, Western Blot and ELISA showed that IKKα-silenced could promote the expression of MASPIN and inhibit the secretion of inflammatory factors; while MASPIN overexpression could inhibit the expression of IKKα and the secretion of inflammatory factors. Conclusion:These results suggest that MASPIN may inhibit the secretion of inflammatory factors through the non-canonical pathway of NF-κB, and high expression of MASPIN is expected to be a potential inhibitor against the canceration of OPMDs.更多还原