HIPK2抑制非小细胞肺癌上皮间质转化及迁移侵袭的作用及机制

doi:10.6040/j.issn.1671-7554.0.2016.227

摘要/Abstract

摘要： 目的研究同源结构域相互作用蛋白激酶2(HIPK2)对非小细胞肺癌(NSCLC)细胞上皮间质转化(EMT)以及迁移侵袭的影响及机制。方法应用Lipofectamine 2000将HIPK2高表达质粒pcDNA3.1-HIPK2和对照质粒pcDNA3.1-Vector转染至A549、H520细胞,分别为高表达组和对照组;应用慢病毒转染HIPK2干扰质粒GV248 shHIPK2和对照质粒GV248 shVector,在A549细胞中分别构建沉默组和对照组;在A549 细胞沉默组中应用Lipofectamine 2000转染GV248 shZEB1质粒和GV248 shVector质粒,分别为干扰组和对照组。Western blotting检测HIPK2表达水平及其对EMT标记蛋白的影响;应用细胞划痕实验、Transwell实验、Matrigel实验检测细胞迁移、侵袭能力。结果与对照组相比,高表达组上皮细胞标记蛋白表达增强(P<0.01)、间质细胞标记蛋白表达下降(P<0.01),EMT受到抑制。高表达组迁移距离小于对照组(P<0.05),高表达组细胞迁移和侵袭至下室的细胞数也明显减少(P<0.01)。沉默HIPK2则促进EMT发生、增强细胞的迁移和侵袭。干扰锌指E盒结合蛋白(ZEB1)的表达可以缓解HIPK2沉默引起的EMT和迁移侵袭现象。结论 HIPK2具有抑制NSCLC细胞EMT和迁移侵袭的作用,其发生机制可能与转录因子ZEB1有关。

关键词: 癌,非小细胞肺, 迁移, 侵袭, 上皮间质转化, 同源结构域相互作用蛋白激酶2

Abstract: Objective To investigate the role of homeodomain interacting protein kinase 2(HIPK2)in epithelial-mesenchymal transition(EMT)and metastasis in non-small cell lung cancer(NSCLC)cells, and to explore the possible mechanism. Methods The A549 and H520 cells were divided into high expression group and control group, then transfected with pcDNA3.1-HIPK2 and pcDNA3.1-Vector with Lipofectamine 2000. HIPK2 silenced and control cell lines were constructed in A549 by the transfection of GV248 shHIPK2 and GV248 shVector with lentivirus. GV248 shZEB1 and GV248 shVector were transfected with Lipofectamine 2000 in A549 cells in which HIPK2 was silenced. The expression of HIPK2 and EMT markers were detected by Western blotting. The migration and invasion ability of cells were detected through wound scratch assay, Transwell assay and Matrigel assay. Results Compared with control group, the expression of epithelial cell markers in high expression group were enhanced(P<0.01), and the expression of mesenchymal cell markers were descended(P<0.01), which repressed the EMT process. Meanwhile, wound scratch assay showed that the migrated distance of high expression group were less than the control group(P<0.05). Transwell assay and Matrigel assay showed that the number of cells that migrated and invaded through the membrane were obviously 山东大学学报 (医学版)54卷11期 -郑荟,等.HIPK2抑制非小细胞肺癌上皮间质转化及迁移侵袭的作用及机制 \=-less than those in control group(P<0.01). Conversely, silencing HIPK2 generated the opposite effects. The enhanced EMT and metastasis induced by HIPK2 silencing could be reversed by interfering zinc finger E-box binding homeobox 1(ZEB1). Conclusion HIPK2 represses EMT, migration and invasion of NSCLC cells, and the possible mechanism is related to transcription factor ZEB1.

Key words: Carcinoma, non-small cell lung, Epithelial-mesenchymal transition, Homeodomain interacting protein kinase 2, Migration, Invasion

中图分类号:

R734.2

郑荟,魏光伟. HIPK2抑制非小细胞肺癌上皮间质转化及迁移侵袭的作用及机制[J]. 山东大学学报（医学版）, 2016, 54(11): 7-12.

ZHENG Hui, WEI Guangwei. Mechanical function of HIPK2 in epithelial-mesenchymal transition and metastasis of non-small cell lung cancer[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(11): 7-12.

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