沉默CRAF基因表达对肝癌细胞侵袭和迁移的影响

doi:10.13481/j.1671-587x.20180312

Abstract

Abstract: Objective: To investigate the effects of high expression of CRAF on the invasion and metastasis of hepatocellular carcinoma cells(HCC),and to clarify their mechanisms. Methods: The SMMC7721 cells with high expression of CRAF were regarded as the object,and the expressions of CRAF in SMMC7721 cells were down-regulated by shRNA.The SMMC7721 cells were divided into control group and silencing groups(shCRAF#1 transfection group and shCRAF#2 transfection group).The wound closure index of SMMC7721 cells was detected by cell scratch test,the number of transmembrane cells of SMMC7721 cells was detected by Transwell assay,and the expression levels of matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9) in SMMC7721 cells were detected by Western blotting method. Results: The scratch test results showed that compared with control group,the wound closure indexes of the SMMC7721 cells in silencing groups were decreased (P<0.05).The Transwell assay results show that compared with control group,the number of transmembrane cells in silencing groups was decreased (P<0.05).The Western blotting results showed that compared with control group,the expression levels of MMP-2 and MMP-9 in the SMMC7721 cells in silencing group were decreased (P<0.05). Conclusion: Silencing CRAF can inhibit the invasion and migration of HCC SMMC7721 cells by inhibiting the expressions of MMP-2 and MMP-9.

Key words: C-Raf proto oncogene serine/threonine protein kinase, liver neoplasms, invasion, metastasis

CLC Number:

R735.7

LIU Xia, QI Fengjie, DU Xiaoyuan. Effects of silencing expression of CRAF gene on invasion and metastasis of hepatocellular carcinoma cells[J].Journal of Jilin University Medicine Edition, 2018, 44(03): 521-525.

References

[1] Wong R,Frenette C. Updates in the management of hepatocellular carcinoma[J]. Gastroenterol Hepatol(N Y),2011,7(1):16-24.
[2] Zhang S,Li J,Liu PG,et al.Pygopus-2 promotes invasion and metastasis of hepatic carcinoma cell by decreasing E-cadherin expression[J].Oncotarget, 2015,6(13):11074-11086.
[3] Chen X,Song B,Lin Y,et al.PTK6 promotes hepatocellular carcinoma cell proliferation and invasion[J].Am J Transl Res,2016,8(10):4354-4361.
[4] 甘声通,胡向阳,陈若冰,等.肝细胞生长因子/c-Met信号通路在肝癌恶性行为中的作用[J].生命的化学,2017,37(3):349-354.
[5] Mitra S,Ghosh B,Gayen N,et al.Bipartite role of heat shock protein 90(Hsp90) keeps CRAF kinase poised for activation[J].J Biol Chem,2016,291(47):24579-24593.
[6] Atefi M,Titz B,Tsoi J,et al.CRAF R391W is a melanoma driver oncogene[J].Sci Rep,2016,6:27454.
[7] Cepeda MA,Evered CL,Pelling JJH,et al.Inhibition of MT1-MMP proteolytic function and ERK1/2 signalling influences cell migration and invasion through changes in MMP-2 and MMP-9 levels[J].J Cell Commun Signal,2017,11(2):167-179.
[8] Chen L,Li M,Li Q,et al.DKK1 promotes hepatocellular carcinoma cell migration and invasion through β-catenin/MMP7 signaling pathway[J].Mol Cancer,2013,12:157.
[9] Yu Y,Song J,Zhang R,et al.Preoperative neutrophil-to-lymphocyte ratio and tumor-related factors to predict microvascular invasion in patients with hepatocellular carcinoma[J].Oncotarget,2017,8(45):79722-79730.
[10] 李雅睿,王梦瑶,卢桂芳,等.DNMT3B基因在肝细胞癌中的表达及其对肝癌细胞增殖和侵袭迁移的影响[J].西安交通大学学报:医学版,2017,38(3):380-385.
[11] Chatelle CV,Hövermann D,Müller A,et al.Optogenetically controlled RAF tocharacterize BRAF and CRAF protein kinase inhibitors[J].Sci Rep,2016,6(3):23713.
[12] Kamata T,Hussain J,Giblett S,et al.BRAF inactivation drives aneuploidy by deregulating CRAF[J].Cancer Res,2010,70(21):8475-8486.
[13] Montagut C,Sharma SV,Shioda T,et al.Elevated CRAF as a potential mechanism of acquired resistance to BRAF inhibition in melanoma[J].Cancer Res,2008,68(12):4853-4861.
[14] Mielgo A,Seguin L,Huang M,et al.A MEK-independent role for CRAF in mitosis and tumor progression[J].Nat Med,2011,17(12):1641-1645.
[15] Sun X,Li J,Sun Y,et al.miR-7 reverses the resistance to BRAFi in melanoma by targeting EGFR/IGF-1R/CRAF and inhibiting the MAPK and PI3K/AKT signaling pathways[J].Oncotarget,2016,7(33):53558-53570.
[16] Lochhead PA,Clark J,Wang LZ,et al.Tumor cells with KRAS or BRAF mutations or ERK5/MAPK7 amplification are not addicted to ERK5 activity for cell proliferation[J].Cell Cycle,2016,15(4):506-518.
[17] Kuliczkowski W,Radomski M,Gąsior M,et al.MMP-2,MMP-9,and TIMP-4 and response to aspirin in diabetic and nondiabetic patients with stable coronary artery disease:A pilot study[J].Biomed Res Int,2017,2017:9352015.
[18] Webb AH,Gao BT,Goldsmith ZK,et al.Inhibition of MMP-2 and MMP-9 decreases cellular migration,and angiogenesis in vitro models of retinoblastoma[J].BMC Cancer,2017,17(1):434.
[19] Shi WZ,Ju JY,Xiao HJ,et al.Dynamics of MMP-9, MMP-2 and TIMP-1 in a rat model of brain injury combined with traumatic heterotopic ossification[J].Mol Med Rep,2017,15(4):2129-2135.
[20] Zhang QY,Li R,Zeng GF,et al.Dihydromyricetin inhibits migration and invasion of hepatoma cells through regulation of MMP-9 expression[J].World J Gastroenterol,2014,20(29):10082-10093.
[21] Roomi MW,Kalinovsky T,Rath M,et al.In vitro modulation of MMP-2 and MMP-9 in pediatric human sarcoma cell lines by cytokines,inducers and inhibitors[J].Int J Oncol,2014,44(1):27-34.

Effects of silencing expression of CRAF gene on invasion and metastasis of hepatocellular carcinoma cells

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