FRMD8抑制肺癌细胞的增殖与迁移

胡金霞 赵玮 李有杰 战军* 谢书阳*

doi:10.16098/j.issn.0529-1356.2016.05.010

PDF(594 KB)

解剖学报 ›› 2016, Vol. 47 ›› Issue (5) : 638-644. DOI: 10.16098/j.issn.0529-1356.2016.05.010

肿瘤生物学

FRMD8抑制肺癌细胞的增殖与迁移

胡金霞¹ 赵玮¹ 李有杰¹ 战军^2*谢书阳^1*

作者信息 +

FRMD8 inhibits amplification and migration of lung cancer cells

HU Jin-xia¹ ZHAO Wei¹ LI You-jie¹ ZHAN Jun^2*XIE Shu-yang^1*

Author information +

文章历史 +

摘要

目的探讨FERM domain-containing protein 8（FRMD8）在肺癌发生发展中的作用。方法用Western blotting检测肺癌组织及相应癌旁正常组织中FRMD8的表达情况；敲低FRMD8后平板克隆形成实验检测细胞克隆形成能力，MTT法检测细胞增殖情况；过表达FRMD8后流式细胞术检测细胞凋亡情况，Western blotting检测过表达FRMD8后某些细凋亡相关蛋白表达变化；过表达及敲低FRMD8后Transwell细胞迁移实验检测对A549细胞迁移能力的影响。结果 1. 与正常组织相比，肺癌组织中FRMD8表达下调（P＜0.05）； 2. FRMD8敲低，细胞克隆形成能力及增殖速度提高； 3. FRMD8可诱导细胞凋亡，并影响细胞凋亡相关基因的表达或活化： FRMD8过表达可下调Caspase-3、8和bax表达，而p53、激活型Caspase-3、8以及bcl-2 的表达上调； 4. FRMD8过表达抑制细胞迁移能力（P＜0.01），而敲低FRMD8则促进细胞迁移（P＜0.001）。结论 FRMD8可抑制肺癌的发生发展，并具有肿瘤抑制子的功能。

Abstract

Objective To investigate the role of FERM domain-containing protein 8 （FRMD8）in lung cancer progression, and explore the mechanism. Methods Western blotting was used to detect expression of FRMD8 in 14 pairs of matched non-involved lung tissue and lung cancer tissues; Colony formation assay and MTT assay were performed for detecting cell proliferation after A549 cells were transfected with siRNAs. Flow cytometry/FACS was applied to detect apoptosis of A549 cells after FRMD8 being up-regulated, Expression of apoptosis-related proteins was evaluated by Western blotting following FRMD8 was up-regulated. A549 cells were transiently transfected with siRNAs or FRMD8 expression vectors, then Transwell migration assay was carried out to test the influence of FRMD8 on cell migration. Results 1. Compared with normal tissues, FRMD8 expression in lung cancer tissues was down-regulated significantly(P＜0.05); 2. Colony formation and MTT assay showed that inhibition of FRMD8 expression by siRNAs promoted proliferation of A549 cells; 3.Over-expression of FRMD8 induced apoptosis and regulated expression of apoptosis-related proteins: Caspase-3, Caspase-8 and Bax were down-regulated, and cleaved Caspase-3, Caspase-8 and bcl-2, p53 were up-regulated; 4. Down-regulation of FRMD8 promoted migration（P＜0.01）. Over-expression of FRMD8 repressed migration of A549 cells（P＜0.001）.Conclusion FRMD8 represses lung cancer progression and may act as a tumor repressor.

导出引用

胡金霞赵玮李有杰战军谢书阳. FRMD8抑制肺癌细胞的增殖与迁移[J]. 解剖学报. 2016, 47(5): 638-644 https://doi.org/10.16098/j.issn.0529-1356.2016.05.010

HU Jin-xia ZHAO Wei LI You-jie ZHAN Jun XIE Shu-yang. FRMD8 inhibits amplification and migration of lung cancer cells[J]. Acta Anatomica Sinica. 2016, 47(5): 638-644 https://doi.org/10.16098/j.issn.0529-1356.2016.05.010

参考文献

［1］Kategava LS,Changkakoty B,Biechele T,et al.Bili inhibits Wnt/beta-catenin signaling by regulating the recruitment of axin to LRP6［J］.PLoS One,2009,4(7):e6129.

［2］Chishti AH, Kim AC, Marfatia SM, et al. The FERM domain: a unique module involved in the linkage of cytoplasmic proteins to the membrane［J］. Trends Biochem Sci, 1998, 23, 281-282.

［3］Frame MC, Patel H, Serrels B,et al.The FERM domain: organizing the structure and function of FAK［J］. Nat Rev Mol Cell Biol, 2010, 11(11): 802-814.

［4］Zhan J, Zhu X, Guo Y, et al. Opposite role of Kindlin-1 and Kindlin-2 in lung cancers［J］. PLoS One,2012,7(11):e50313.

［5］Haase D, Meister M, Muley T,et al. FRMD3, a novel putative tumour suppressor in NSCLC［J］. Oncogene,2007,26(30): 4464-4468.

［6］Hu J, Niu M, Li X,et al.FERM domain-containing protein FRMD5 regulates cll motility via binding to integrin beta5 subunit and ROCK1［J］.FEBS Lett,2014,588(23):4348-4355.

［7］Wang T, Pei X, Zhan J, et al. FERM-containing protein FRMD5 is a p120-catenin interacting protein that regulates tumor progression［J］. FEBS Lett, 2012,586(19): 3044-3050.

［8］Galvan A, Falvella FS, Frullanti E, et al. Genome-wide association study in discordant sibships identifies multiple inherited susceptibility alleles linked to lung cancer［J］. Carcinogenesis, 2010,31(3): 462-465.

［9］Longobardi Iotti G, Di Rosa P, et al. Prep1 (pKnox1)-deficiency leads to spontaneous tumor development in mice and accelerates EmuMyc lymphomagenesis: a tumor suppressor role for Prep1［J］. Mol Oncol, 2010, 4(2): 126-134.

［10］Lim ST, Chen XL, Lim Y, et al. Nuclear FAK promotes cell proliferation and survival through FERM-enhanced p53 degradation［J］. Mol Cell,2008(1):29, 9-22.

［11］Hamaratoglu F, Willecke M, Kango-Singh M,et al. The tumour-suppressor genes NF2/Merlin and Expanded act through Hippo signalling to regulate cell proliferation and apoptosis［J］. Nat Cell Biol,2006,8(1): 27-36.

［12］Wang L, Deng W, Shi T, et al. URP2SF, a FERM and PH domain containing protein, regulates NF-kappaB and apoptosis［J］. Biochem Biophys Res Commun, 2008,368(4): 899-906.

［13］Jakel H, Weinl C, Hengst L, et al. Phosphorylation of p27Kip1 by JAK2 directly links cytokine receptor signaling to cell cycle control［J］. Oncogene,2011,30(32): 3502-3512.

［14］Lim ST, Miller NL, Nam JO,et al. Pyk2 inhibition of p53 as an adaptive and intrinsic mechanism facilitating cell proliferation and survival［J］. J Biol Chem,2010,285(3): 1743-1753.

［15］Wang Z, Ma B, Li H, et al. Protein 4-1N acts as a potential tumor suppressor linking PP1 to JNK-c-Jun pathway regulation in NSCLC［J］. Oncotarget, 2016,(1): 509-523.

［16］Yang M, Chen X，Zhang ShW,et al, Research progress of biological function in integrin-interacting protein kindlin family［J］.Acta Anatomica Sinica,2014,45(6):865-869. (in Chinese)

杨玫，陈曦，张水文,等，整合素相互作用蛋白Kindlin 家族的生物学功能研究进展［J］.解剖学报，2014,45(6): 865-869.

［17］Hyduk SJ, Rullo J, Cano AP, et al.Talin-1 and kindlin-3 regulate alpha4beta1 integrin-mediated adhesion stabilization, but not G protein-coupled receptor-induced affinity upregulation［J］. J Immunol, 2011,187(8): 4360-4368.

［18］Pinon P, P?rssinen J, Vazquez P, et al. Talin-bound NPLY motif recruits integrin-signaling adapters to regulate cell spreading and mechanosensing［J］. J Cell Biol, 2014, 205(2): 265-281.

［19］Melis M,Cau M,Corraine S, et al.Cerebral cavernous malformations and unilateral moyamoya in a patient with a new mutation in the KRIT-1/CCM1 gene［J］.Cerebrovasc Dis, 2014, 38(4):311-312.

［20］Han M, Nagele E, DeMarshall C,et al.Diagnosis of Parkinson’s disease based on disease-specific autoantibody profiles in human sera［J］. PLoS One, 2012, 7(2): e32383.

［21］Santiago JA, Potashkin JA.Current challenges towards the development of a blood test for Parkinson’s,disease［J］. Diagnostics(Basel),2014,4(4) :153-164.