赖氨大黄酸通过升高miRNA-451抑制肺癌A549细胞迁移  

Rhein lysinate inhibits migration in lung cancer A549 cells by miRNA-451 upregulation

作　　者：纪春梅 甄永占[2] 骆广玲[3] 赵毓芳[2] 朱丽华[2] 

机构地区：[1]唐山协和医院呼吸内科,河北唐山063000 [2]河北联合大学基础医学院组织学与胚胎学教研室,河北唐山063000 [3]开滦医疗集团林西医院妇产科,河北唐山063000

出　　处：《第三军医大学学报》2014年第3期248-252,共5页Journal of Third Military Medical University

基　　金：河北省自然科学基金(H2012401030);国家自然科学基金(81001439)~~

摘　　要：目的研究赖氨大黄酸(rhein lysinate,RHL)对肺癌细胞株(A549)迁移的影响及miRNA-451在其中的作用。方法选取处于对数生长期的A549细胞株,并分为对照组、不同剂量(0、10、20、40、80、160μmol/L)的RHL组。采用MTT法检测各组细胞48 h增殖情况;细胞划痕实验和Transwell实验检测各组细胞48 h迁移情况;Real-time PCR检测各组细胞48 h miRNA-451和基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)的转录水平;Western blot检测各组细胞48 h MMP-9蛋白水平变化。结果细胞培养48 h后,RHL处理组A549细胞增殖活性呈现明显剂量依赖性抑制。细胞划痕实验和Transwell实验均发现80μmol/L RHL能够抑制A549细胞迁移。同时80μmol/L以上剂量的RHL能够显著升高miR-451水平,降低MMP-9的转录水平(P<0.05)。Western blot检测结果也证实了80μmol/L以上剂量的RHL能够显著抑制MMP-9蛋白表达水平(P<0.05)。结论 RHL通过诱导miR-451的表达、抑制MMP-9的表达,从而抑制A549细胞迁移。Objective To investigate the effect of Rhein lysinate （RHL） on the migration of human lung cancer cell line （A549） and the role of miRNA-451 signal pathway in cell migration, and to provide theoretical foundation for clinical trial of RHL against lung cancer. Methods A549 cells in logarithm growth phase were selected and randomly divided into various groups： a blank control group and different concentra- tions of RHL groups. After A549 cells were treated by RHL for 48 h, MrFF assay was used to detect A549 cell proliferation. Wound healing and Transwell assay were used to detect the migratory ability of A549 cells. The levels of miRNA-451 and MMP-9 mRNA were detected by real-time PCR. The expression of MMP-9 protein was detected by Western blot. Results After A549 cells were treated by RHL for 48 h, cell viability was inhibited in a dose-dependent manner. Wound healing and Transwell assay results showed that RHL （80 μmol/L） could inhibit the migratory ability of A549 cells. Compared with control group, the levels of miR-451 were increased, while the mRNA levels of MMP-9 were decreased in RHL （ ≥ 80μmol/L） -treated groups （ P 〈 0. 05 ）. Compared with control group, Western blot results also showed that the expression of MMP-9 protein was decreased in RHL （ ≥80 μmol/L）-treated group （P 〈0. 05）. Conclusion RHL can inhibit the migratory ability of A549 cells by inducing the expression of miR-451 and thereby inhibiting the transcription and expression of MMP-9.

关 键 词：赖氨大黄酸 肺肿瘤 细胞迁移 miRNA-451 基质金属蛋白酶-9 

分 类 号：R73-361[医药卫生—肿瘤] R734.2[医药卫生—临床医学]