摘要
目的探讨中介因子复合体(mediator 27,MED27)在肺癌组织样本和肺癌细胞中的表达并进一步观察MED27在肺癌细胞中的生物学功能。方法采用免疫组化法和Western blot法检测MED27在70例肺癌组织以及5种不同肺癌细胞中的表达,分析MED27蛋白表达与肺癌临床病理特征的相关性;设计沉默MED27基因的siRNA,利用Western blot法检测MED27siRNA在肺癌细胞中的沉默效率;CCK-8法检测细胞增殖能力的变化,划痕和Transwell实验评估细胞的迁移和侵袭能力的变化;Western blot法检测迁移侵袭相关蛋白的变化。结果免疫组化和Western blot检测结果表明,MED27在肺癌组织以及肺癌细胞系中的表达水平明显上调(P<0.05)。MED27表达与淋巴结转移呈正相关(χ2=9.438,P=0.002,P<0.05),与患者性别、年龄、肿瘤T分期、远处转移等均无相关性(P>0.05);利用小干扰RNA沉默MED27的表达,可以抑制H460细胞的增殖、迁移和侵袭(P<0.05)。同时,迁移相关蛋白MMP-2和MMP-9表达明显下调,侵袭相关蛋白E-cadherin表达也明显下调,而E-cadherin的负性调控蛋白Snail表达升高。结论 MED27在肺癌组织和细胞系中高表达,且MED27阳性肺癌患者预后更差。沉默MED27的表达可以抑制肺癌细胞的增殖、迁移和侵袭能力,以上结果提示MED27可以作为临床肺癌基因治疗的潜在靶标。
Purpose To investigate the expression level of MED27 in lung cancer tissue samples and lung cancer cell lines and to further study the biological function of MED27 in lung cancer cells. Methods Immunohistochemistry and Western blot were used to detect MED27 expression in 70 lung cancer tissues and 5 different lung cancer cell lines, and the correlation between MED27 expression and gender, age as well as PTNM was also analyzed. The silence sequence of MED27 was designed by the siRNA technique. Western blot was used to detect the silence efficiency of MED27. The proliferation, migration and invasion ability of cells were assessed by CCK-8 assay, Scratch assay and Transwell assay after the MED27 was knocked down. Western blot was used to detect the expression of protein involved in the cell proliferation, migration and invasion. Results The results of immunohistochemistry and Western blot showed that MED27 expression was higher in lung cancer tissues and cells （ P 〈 0. 05 ）. The expression of MED27 was positively correlated with lymph node metastasis （ X2 = 9. 438, P = 0. 002, P 〈 0. 05 ）. However, it was not related with gender, age, tumor size and distant metastasis （ P 〉 0. 05 ）. The knockdown of MED27 by MED27 specific siRNA could inhibit the prolifera- tion, migration and invasion of H460 cells （ P 〈 0.05 ）. The expression of MMP-2 and MMP-9 involved in the cell migration that were significantly inhibited in H460 cells transfected by MED27 siRNA, and the expression of E-cadherin, related with cell invasion was also decreased, while E-cadherin negative regulatory protein Snail was increased. Conclusion MED27 is highly expressed in lung cancer tissues and cells and high expression of MED27 predicts poor prognosis in lung cancer patients. The knockdown of MED27 inhibits the proliferation, migration and invasion ability of lung cancer cells. All of the above results suggest that MED27 is expected to be a candidate target of lung cancer gene therapy.
出处
《临床与实验病理学杂志》
CSCD
北大核心
2017年第10期1086-1091,共6页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学青年基金(81702831
81402139)
