长链非编码RNA LINC00473对人胃癌细胞增殖和迁移能力的影响
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摘要
目的探讨LINC00473在人胃癌细胞系中的表达水平及其对胃癌细胞生物学行为的影响。方法采用实时荧光定量PCR检测胃癌细胞中LINC00473的表达水平;转染靶向LINC00473的siRNA片段或LINC00473过表达载体以构建敲减或过表达的胃癌细胞系;通过CCK-8细胞增殖实验、平板克隆形成实验、Transwell迁移实验、western blot实验检测细胞的增殖和迁移能力以及EMT相关蛋白标志物表达水平的变化。结果与GES-1细胞相比,LINC00473在胃癌细胞中的表达水平明显降低(P<0.05)。与siNC转染对照组相比,敲减LINC00473表达后,靶细胞的增殖能力(F=163.10,P<0.01)、克隆形成(t=3.29,P<0.05)和迁移能力(t=4.68,P<0.05)明显增加;E-cadherin表达减弱(t=4.08,P<0.05),N-cadherin(t=5.06,P<0.01)、Snail(t=7.69,P<0.01)和Vimentin(t=3.82,P<0.05)蛋白的表达水平升高。与Vector转染对照组相比,上调LINC00473表达后靶细胞的增殖能力(F=186.00,P<0.01)、克隆形成(t=3.22,P<0.05)和迁移能力(t=5.52,P<0.01)明显下降;E-cadherin表达升高(t=2.90,P<0.05),N-cadherin(t=7.44,P<0.01)、Snail(t=2.78,P<0.05)和Vimentin(t=4.64,P<0.01)蛋白的表达水平降低。结论敲减LINC00473的表达可促进胃癌细胞的增殖、迁移的能力,可能通过调控EMT参与胃癌细胞的迁移。
Objective To investigate the expression profiles of long non-coding RNA LINC00473 in gastric cancer cells and its effects on the proliferation and migration of gastric cancer cells. Methods The expression level of LINC00473 in gastric cancer cells was verified by qRT-PCR system. LINC00473 siRNA segment and overexpression vector were separately transfected into gastric cancer cells by the method of lipofection. The proliferation and migration abilities of gastric cancer cells with LINC00473 knockdown or overexpression in vitro were evaluated by cell counting kit-8(CCK8) assay, colony formation assay and Transwell migration assay. The expression levels of proteins involved in epithelial-mesenchymal transition(EMT) were examined by western blot analysis. Results The expression levels of LINC00473 were decreased in gastric cancer cells compared with that in human gastric epithelial cell strain GES-1(P<0.05). LINC00473 knockdown cells showed significant increased ability for cell growth(F=163.10, P<0.01) and colony formation(t=3.29, P<0.05) compared with the knockdown cells in scramble control. The results of Transwell migration assay showed that LINC00473-knockdown enhanced the migratory abilities of gastric cancer cells(t=4.68, P<0.05). The knockdown of LINC00473 downregulated E-cadherin expression(t=4.08, P<0.05) and upregulated N-cadherin(t=5.06, P<0.01), Snail(t=7.69, P<0.01) and Vimentin(t=3.82, P<0.05) expression. Compared with the control group, LINC00473 overexpression cells showed significantly decreased cell growth(F=186.00, P<0.01) and colony formation ability(t=3.22, P<0.05). The results of Transwell migration assay showed that LINC00473-overexpression reduced the migratory ability of gastric cancer cells(t=5.52, P<0.05). The overexpression of LINC00473 enhanced E-cadherin expression(t=2.90, P<0.05) and reduced the expressions of N-cadherin(t=7.44, P<0.01), Snail(t=2.78, P<0.05) and Vimentin(t=4.64, P<0.01). Conclusion The knockdown of LINC00473 may promote gastric cancer cell proliferation and migration in vitro by regulating EMT.
Objective To investigate the expression profiles of long non-coding RNA LINC00473 in gastric cancer cells and its effects on the proliferation and migration of gastric cancer cells. Methods The expression level of LINC00473 in gastric cancer cells was verified by qRT-PCR system. LINC00473 siRNA segment and overexpression vector were separately transfected into gastric cancer cells by the method of lipofection. The proliferation and migration abilities of gastric cancer cells with LINC00473 knockdown or overexpression in vitro were evaluated by cell counting kit-8(CCK8) assay, colony formation assay and Transwell migration assay. The expression levels of proteins involved in epithelial-mesenchymal transition(EMT) were examined by western blot analysis. Results The expression levels of LINC00473 were decreased in gastric cancer cells compared with that in human gastric epithelial cell strain GES-1(P<0.05). LINC00473 knockdown cells showed significant increased ability for cell growth(F=163.10, P<0.01) and colony formation(t=3.29, P<0.05) compared with the knockdown cells in scramble control. The results of Transwell migration assay showed that LINC00473-knockdown enhanced the migratory abilities of gastric cancer cells(t=4.68, P<0.05). The knockdown of LINC00473 downregulated E-cadherin expression(t=4.08, P<0.05) and upregulated N-cadherin(t=5.06, P<0.01), Snail(t=7.69, P<0.01) and Vimentin(t=3.82, P<0.05) expression. Compared with the control group, LINC00473 overexpression cells showed significantly decreased cell growth(F=186.00, P<0.01) and colony formation ability(t=3.22, P<0.05). The results of Transwell migration assay showed that LINC00473-overexpression reduced the migratory ability of gastric cancer cells(t=5.52, P<0.05). The overexpression of LINC00473 enhanced E-cadherin expression(t=2.90, P<0.05) and reduced the expressions of N-cadherin(t=7.44, P<0.01), Snail(t=2.78, P<0.05) and Vimentin(t=4.64, P<0.01). Conclusion The knockdown of LINC00473 may promote gastric cancer cell proliferation and migration in vitro by regulating EMT.
引文
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[4]Chen Z,Lin S,Li JL,et al.CRTC1-MAML2 fusion-induced lncRNA LINC00473 expression maintains the growth and survival of human mucoepidermoid carcinoma cells[J].Oncogene,2018,37(14):1885-1895.
[5]Han PB,Ji XJ,Zhang M,et al.Upregulation of lncRNA LINC00473 promotes radioresistance of HNSCC cells through activating Wnt/β-catenin signaling pathway[J].Eur Rev Med Pharmacol Sci,2018,22(21):7305-7313.
[6]Shi C,Yang Y,Yu J,et al.The long noncoding RNA LINC00473,a target of microRNA 34a,promotes tumorigenesis by inhibiting ILF2 degradation in cervical cancer[J].Am J Cancer Res,2017,7(11):2157-2168.
[7]Zhu S,Fu W,Zhang L,et al.LINC00473 antagonizes the tumour suppressor miR-195 to mediate the pathogenesis of Wilms tumour via IKKα[J].Cell Prolif,2018,51(1).doi:10.1111/cpr.12416.
[8]Zhang W,Song Y.LINC00473 predicts poor prognosis and regulates cell migration and invasion in gastric cancer[J].Biomed Pharmacother,2018,107:1-6.
[9]Yu H,Rong L.Emerging role of long non-coding RNA in the development of gastric cancer[J].World J Gastrointest Oncol,2018,10(9):260-270.
[10]曹季军,王金湖,申娴娟,等.lncRNA在胃癌中作用的研究进展[J].临床检验杂志,2017,35(6):448-451.