EMT分子标志物在肝癌细胞系中的表达及其意义
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摘要
目的探讨人肝癌细胞系EMT相关分子标志物的表达情况,为确定相关的肝癌细胞研究模型奠定基础。方法利用实时定量PCR方法检测5株人肝癌细胞系(SNU368、SNU739、HLF、Huh-1和MHCC97L)中E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的mRNA表达,Western blot和免疫荧光检测E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的蛋白表达。结果 E-cadherin、ZO-1、N-cadherin、Vimentin、Snail和Slug的mRNA在5种肝癌细胞中均有不同程度表达, E-cadherin、ZO-1、N-cadherin和Slug蛋白在5个细胞株中亦有表达,Vimentin蛋白主要在SNU368、SNU739、HLF和MHCC97L细胞中表达明显,Snail蛋白在SNU368,SNU739,Huh-1和MHCC97L细胞中表达较高。结论 HLF细胞系表现出更明显的间质细胞表型特点,Huh-1和SNU368则表现为较典型的上皮细胞表型特点。
Objective To investigate the expressions of molecular markers associated with epithelial-mesenchymal transition(EMT) in human hepatocellular carcinoma(HCC) cells so as to lay foundation for determining related HCC cells model. Methods We detected the mRNA and protein expression levels of EMT-associated genes, including E-cadherin, ZO-1, N-cadherin, Vimentin, Snail and Slug in five HCC cell lines, using quantitative real-time polymerase chain reaction analysis, Western blot and immunofluorescence. Results The mRNA of EMT-associated genes was expressed in all of the five cell lines. The expressions of E-cadherin, ZO-1, N-cadherin and Slug protein were detected in all of the five cell lines. Vimentin protein was found in SNU368, SNU739, HLF and MHCC97 L cells. Snail protein was detected in SNU368, SNU739, Huh-1 and MHCC97 L cells. Conclusion HLF seems to display the phenotype of mesenchymal cells whereas Huh-1 and SNU368 remain the epithelial phenotype.
Objective To investigate the expressions of molecular markers associated with epithelial-mesenchymal transition(EMT) in human hepatocellular carcinoma(HCC) cells so as to lay foundation for determining related HCC cells model. Methods We detected the mRNA and protein expression levels of EMT-associated genes, including E-cadherin, ZO-1, N-cadherin, Vimentin, Snail and Slug in five HCC cell lines, using quantitative real-time polymerase chain reaction analysis, Western blot and immunofluorescence. Results The mRNA of EMT-associated genes was expressed in all of the five cell lines. The expressions of E-cadherin, ZO-1, N-cadherin and Slug protein were detected in all of the five cell lines. Vimentin protein was found in SNU368, SNU739, HLF and MHCC97 L cells. Snail protein was detected in SNU368, SNU739, Huh-1 and MHCC97 L cells. Conclusion HLF seems to display the phenotype of mesenchymal cells whereas Huh-1 and SNU368 remain the epithelial phenotype.
引文
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[14] VEGA S,MORALES AV,OCANA OH,et al.Snail blocks the cell cycle and confers resistance to cell death[J].Genes Dev,2004,18(10):1131-1143.
[15] AOMATSU K,ARAO T,ABE K,et al.Slug is upregulated during wound healing and regulates cellular phenotypes in corneal epithelial cells[J].Invest Ophth Vis Sci,2012,53(2):751-756.
[16] WELCH-REARDON KM,EHSAN SM,WANG KH,et al.Angiogenic sprouting is regulated by endothelial cell expression of Slug[J].J Cell Sci,2014,127(9):2017-2028.
[2] NIETO MA.Epithelial plasticity:A common theme in embryonic and cancer cells[J].Science,2013,342(6159):708-713.
[3] PUISIEUX A,BRABLETZ T,CARAMEL J.Oncogenic roles of EMT-inducing transcription factors[J].Nat Cell Bio,2014,16(6):488-494.
[4] PEINADO H,OLMEDA D,CANO A.Snail,ZEB and bHLH factors in tumour progression:An alliance against the epithelial phenotype[J].Nat Rev Cancer,2007,7(6):415-428.
[5] COOPMAN P,DJIANE A.Adherens junction and E-cadherin complex regulation by epithelial polarity[J].Cell Mol Life Sci,2016,73(18):3535-3553.
[6] FOUNOUNOU N,LOYER N,LE BORGNE R.Septins regulate the contractility of the actomyosin ring to enable adherens junction remodeling during cytokinesis of epithelial cells[J].Dev Cell,2013,24(3):242-255.
[7] GIANNELLI G,KOUDELKOVA P,DITURI F,et al.Role of epithelial to mesenchymal transition in hepatocellular carcinoma[J].J Hepatol,2016,65(4):798-808.
[8] VAN ZIJL F,MALL S,MACHAT G,et al.A Human model of epithelial to mesenchymal transition to monitor drug efficacy in hepatocellular carcinoma progression[J].Mol Cancer Ther,2011,10(5):850-860.
[9] NAGAI T,ARAO T,FURUTA K,et al.Sorafenib inhibits the hepatocyte growth factor-mediated epithelial mesenchymal transition in hepatocellular carcinoma[J].Mol Cancer Ther,2011,10(1):169-177.
[10] XIAO S,CHANG RM,YANG MY,et al.Actin-like 6A predicts poor prognosis of hepatocellular carcinoma and promotes metastasis and epithelial-mesenchymal transition[J].Hepatology,2016,63(4):1256-1271.
[11] YANG MH,CHEN CL,CHAU GY,et al.Comprehensive analysis of the independent effect of Twist and Snail in promoting metastasis of hepatocellular carcinoma[J].Hepatology,2009,50(5):1464-1474.
[12] GIANNELLI G,BERGAMINI C,FRANSVEA E,et al.Laminin-5 with transforming growth factor-beta1 induces epithelial to mesenchymal transition in hepatocellular carcinoma[J].Gastroenterology,2005,129(5):1375-1383.
[13] DING W,YOU HN,DANG H,et al.Epithelial-to-mesenchymal transition of murine liver tumor cells promotes invasion[J].Hepatology,2010,52(3):945-953.
[14] VEGA S,MORALES AV,OCANA OH,et al.Snail blocks the cell cycle and confers resistance to cell death[J].Genes Dev,2004,18(10):1131-1143.
[15] AOMATSU K,ARAO T,ABE K,et al.Slug is upregulated during wound healing and regulates cellular phenotypes in corneal epithelial cells[J].Invest Ophth Vis Sci,2012,53(2):751-756.
[16] WELCH-REARDON KM,EHSAN SM,WANG KH,et al.Angiogenic sprouting is regulated by endothelial cell expression of Slug[J].J Cell Sci,2014,127(9):2017-2028.