长链非编码RNA ZEB1-AS1上调肿瘤细胞SGC-7901和NCI-N87受体酪氨酸激酶c-MET蛋白表达水平
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摘要
目的探讨长链非编码RNA(lncRNA)ZEB1-AS1在肿瘤细胞中与受体酪氨酸激酶c-MET的作用关系。方法在胃癌SGC-7901和NCI-N87细胞中转染pcDNA3.0-ZEB1-AS1质粒,采用CCK-8实验检测其对细胞增殖的影响,流式细胞术检测其对细胞周期和细胞凋亡的影响,实时荧光定量PCR(qRT-PCR)和Western印迹法分析其对c-MET mRNA和蛋白质表达水平的影响。结果 lncRNA ZEB1-AS1促进胃癌SGC-7901和NCI-N87细胞增殖,对细胞周期和凋亡无明显影响;过表达lncRNA ZEB1-AS1上调SGC-7901和NCI-N87细胞中受体酪氨酸激酶c-MET蛋白水平。结论 lncRNA ZEB1-AS1与原癌基因受体酪氨酸激酶在肿瘤细胞中存在相互调控的关系,为阐明受体酪氨酸激酶的机制提供了新线索。
Objective To investigate the relationships between long noncoding RNA(lncRNA)zinc finger E-box binding homeobox 1 antisense 1(ZEB1-AS1)and receptor tyrosine kinase(RTK)c-MET in tumor cells.Methods The plasmid pcDNA3.0-ZEB1-AS1 was transfected into SGC-7901 and NCI-N87 cells respectively.CCK-8 assay was used to detect its effect on cell proliferation,while flow cytometry was used to analyze its effect on cell cycle and cell apoptosis.The mRNA and protein expression levels of c-MET in SGC-7901 and NCI-N87 cells were analyzed by qRT-PCR and Western blotting.Results Overexpression of lncRNA ZEB1-AS1 promoted proliferation of gastric tumor cells,SGC-7901 and NCI-N87,but had no significant effect on cell cycle and apoptosis.However,overexpression of lncRNA ZEB1-AS1 upregulat-ed RTK c-MET protein levels in SGC-7901 and NCI-N87 cells.ConclusionlncRNA ZEB1-AS1 and RTK c-MET as proto-oncogenes have a mutual regulatory relationship in tumor cells.The above results provide new clues to the mechanism of RTK in cancer.
Objective To investigate the relationships between long noncoding RNA(lncRNA)zinc finger E-box binding homeobox 1 antisense 1(ZEB1-AS1)and receptor tyrosine kinase(RTK)c-MET in tumor cells.Methods The plasmid pcDNA3.0-ZEB1-AS1 was transfected into SGC-7901 and NCI-N87 cells respectively.CCK-8 assay was used to detect its effect on cell proliferation,while flow cytometry was used to analyze its effect on cell cycle and cell apoptosis.The mRNA and protein expression levels of c-MET in SGC-7901 and NCI-N87 cells were analyzed by qRT-PCR and Western blotting.Results Overexpression of lncRNA ZEB1-AS1 promoted proliferation of gastric tumor cells,SGC-7901 and NCI-N87,but had no significant effect on cell cycle and apoptosis.However,overexpression of lncRNA ZEB1-AS1 upregulat-ed RTK c-MET protein levels in SGC-7901 and NCI-N87 cells.ConclusionlncRNA ZEB1-AS1 and RTK c-MET as proto-oncogenes have a mutual regulatory relationship in tumor cells.The above results provide new clues to the mechanism of RTK in cancer.
引文
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[3]Fatica A,Bozzoni I.Long non-coding RNAs:new players in cell differentiation and development[J].Nat Rev Genet,2014,15(1):7-21.
[4]Zhou Y,Zhang X,Klibanski A.MEG3 noncoding RNA:a tumor suppressor[J].J Mol Endocrinol,2012,48(3):R45-R53.
[5]Liu C,Tang DG.MicroRNA regulation of cancer stem cells[J].Cancer Res,2011,71(18):5950-5954.
[6]Wang Y,Hou J,He D,et al.The emerging function and mechanism of ceRNAs in cancer[J].Trends Genet,2016,32(4):211-224.
[7]Wang S,Liang K,Hu Q,et al.JAK2-binding long noncoding RNA promotes breast cancer brain metastasis[J].J Clin Invest,2017,127(12):4498-4515.
[8]Du Z,Sun T,Hacisuleyman E,et al.Integrative analyses reveal a long noncoding RNA-mediated sponge regulatory network in prostate cancer[J].Nat Commun,2016,7:10982.
[9]Li T,Xie J,Shen C,et al.Upregulation of long noncoding RNAZEB1-AS1 promotes tumor metastasis and predicts poor prognosis in hepatocellular carcinoma[J].Oncogene,2016,35(12):1575-1584.
[10]Wang YL,Bai Y,Yao WJ,et al.Expression of long non-coding RNA ZEB1-AS1 in esophageal squamous cell carcinoma and its correlation with tumor progression and patient survival[J].Int JClin Exp Med,2015,8(9):11871.
[11]Liu C,Lin J.Long noncoding RNA ZEB1-AS1 acts as an onco-gene in osteosarcoma by epigenetically activating ZEB1[J].Am J Transl Res,2016,8(10):4095-4105.
[12]Lin J,Zhan Y,Liu Y,et al.Increased expression of ZEB1-AS1 correlates with higher histopathological grade and promotes tu-morigenesis in bladder cancer[J].Oncotarget,2017,8(15):24202-24212.
[13]Wang Q,Du XL,Yang M,et al.LncRNA ZEB1-AS1 contrib-utes to STAT3 activation by associating with IL-11 in B-lympho-blastic leukemia[J].Biotechnol Lett,2017,39(12):1801-1810.
[14]Liu XJ,Li SL,Li JS,et al.Long non-coding RNA ZEB1-AS1 is associated with poor prognosis in gastric cancer and promotes cancer cell metastasis[J].Eur Rev Med Pharmacol Sci,2018,22(9):2624-2630.
[15]Maroun CR,Rowlands T.The Met receptor tyrosine kinase:a key player in oncogenesis and drug resistance[J].Pharmacol Ther,2014,142(3):316-338.
[16]Lai AZ,Abella JV,Park M.Crosstalk in Met receptor oncogene-sis[J].Trends Cell Biol,2009,19(10):542-555.
[17]郑荣寿,孙可欣,张思维,等.2015年中国恶性肿瘤流行情况分析[J].中华肿瘤杂志,2019,4(1):19-28.
[18]Ji P,Diederichs S,Wang W,et al.MALAT1,a novel noncoding RNA,and thymosin 4 predict metastasis and survival in early-stage non-small cell lung cancer[J].Oncogene,2003,22(39):8031-8041.
[19]Li J,Gao J,Tian W,et al.Long non-coding RNA MALAT1drives gastric cancer progression by regulating HMGB2 modu-lating the miR-1297[J].Cancer Cell Int,2017,17:44.
[20]Yang Z,Xie Q,Chen Z,et al.Resveratrol suppresses the invasion and migration of human gastric cancer cells via inhibition of MALAT1-mediated epithelial-to-mesenchymal transition[J].Exp Ther Med,2019,17(3):1569-1578.
[21]Xi Z,Si J,Nan J.LncRNA MALAT1 potentiates autophagy-associated cisplatin resistance by regulating the microRNA-30b/autophagy-related gene 5 axis in gastric cancer[J].Int J Oncol,2019,54(1):239-248.
[22]Hu X,Feng Y,Zhang D,et al.A functional genomic approach identifies FAL1 as an oncogenic long noncoding RNA that associates with BMI1 and represses p21 expression in cancer[J].Cancer Cell,2014,26(3):344-357.
[23]Zhao L,Ji G,Le X,et al.Long noncoding RNA LINC00092acts in cancer-associated fibroblasts to drive glycolysis and pro-gression of gastric cancer[J].Cancer Res,2017,77(6):1369-1382.
[24]Li H,Wang Y.Long noncoding RNA(lncRNA)MIR22HG suppresses gastric cancer progression through attenuating NOTCH2 signaling[J].Med Sci Monit,2019,25:656-665.
[25]Wu F,Gao H,Liu K,et al.The lncRNA ZEB2-AS1 is upregulated in gastric cancer and affects cell proliferation and invasion via miR-143-5p/HIF-1αaxis[J].Onco Targets Ther,2019,12:657-667.
[26]Jia J,Zhan D,Li J,et al.The contrary functions of lncRNA HO-TAIR/miR-17-5p/PTEN axis and Shenqifuzheng injection on chemosensitivity of gastric cancer cells[J].J Cell Mol Med,2019,23(1):656-669.
[27]Yap TA,Sandhu SK,Alam SM,et al.HGF/c-MET targeted therapeutics:novel strategies for cancer medicine[J].Curr Drug Targets,2011,12(14):2045-2058.
[28]Eder JP,Vande Woude GF,Boerner SA,et al.Novel therapeu-tic inhibitors of the c-Met signaling pathway in cancer[J].Clin Cancer Res,2009,15(7):2207-2214.