背景与目的:簇集素蛋白(clusterin,CLU)在肺癌的发生、发展中发挥重要作用。本研究用分泌型CLU体外处理肺癌细胞株,旨在探讨分泌型CLU对肺癌细胞株的影响。方法:用真核表达纯化的分泌型CLU处理非小细胞肺癌细胞株H460、A549,采用Boydene小室试验检测细胞的迁移能力,用CCK8检测细胞的增殖能力,利用芯片技术分析CLU处理H460细胞的microRNA表达谱,然后采用实时荧光定量PCR检测分泌型CLU处理H460细胞中microRNA的表达,初步分析分泌型CLU执行肿瘤生物学功能的microRNA相关分子机制。结果:分泌型CLU处理组H460迁移的细胞是BSA处理组的3.10倍(P〈0.0001);分泌型CLU处理组A549迁移的细胞是BSA处理组的3.40倍(P〈0.0001);分泌型CLU抑制了肺癌细胞株H460/A549的生长(P〈O.0001);利用芯片技术发现microRNA-302b-3p、microRNA-23a-5p和gmicroRNA-101-5p高表达。结论:在非小细胞肺癌细胞株H460/A549中,真核表达的分泌型CLU促进了细胞迁移、抑制了细胞生长,并且能引起microRNA表达谱的改变,本研究为探索分泌型CLU在非小细胞肺癌中的作用提供了重要的实验依据。
Background and purpose: Primary study showed that clusterin was associated with tumorigenicity. The goal of the study is to investigate the role of secretory clusterin in non-small cell lung cancer cell A549/H460. Methods: The lung cancer cell A549/H460 was treated with purified secretory clusterin, the Boyden Chamber migration assay was used to detected the migration of the lung cancer cell; the CCK8 assay was used to detected the growth of the ceils; microRNA expression spectrum in H460 treated with secretory clusterin was analyzed, after that, we used the real-time florescence quantification detected the expression of microRNA in H460, and the biological function of microRNA molecular mechanisms of secretory clusterin was analyzed. Results: Secretory clusterin promoted the migration in A549/H460 (P〈0.000 1); Secretory clusterin inhibited the growth in H460/A549 (P〈0.000 1); MicroRNA-302b-3p, microRNA-23a-5p and microRNA-101-5p was overexpressed in H460 when treated with secretory clusterin. Conclusion: Secretory clusterin could promote the migration and inhibit the growth of lung cancer cell; It could change the microRNA expression spectrum as well. Our studies revealed that secretory clusterin could be used as a tool for further study, and it is a potential target in the treatment of lung cancer.