外源性α1-抗胰蛋白酶对非小细胞肺癌转移与侵袭的促进作用研究
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摘要
目的探讨α1-抗胰蛋白酶(A1AT)对非小细胞肺癌(NSCLC)转移与侵袭的促进作用。方法向细胞培养液中加入不同质量浓度A1AT(0,600,1 200,2 400ng/mL),采用免疫印迹法和免疫荧光技术分别检测上皮细胞标志物E-cadherin、间质细胞标志物N-cadherin、Vimentin;采用细胞transwell实验和划痕试验检测细胞的转移和侵袭能力。结果 A1AT质量浓度越高,E-cadherin表达越低,N-cadherin和Vimentin表达含量越高,细胞的转移和侵袭能力越强。结论外源性A1AT能够促进NSCLC的转移与侵袭。
Objective To investigate the effect of alpha 1-antitrypsin(A1 AT)on the promotion of metastasis and invasion of non-small cell lung cancer(NSCLC).Methods The different concentrations of A1 AT(0,600,1200,2400 ng/ml)were added into the cell culture solution.Western blot and immunofluorescence were used to detect the epithelial cell marker E-cadherin and the mesenchymal markers including N-cadherin and Vimentin.The transwell and wound-healing assays were used to detect the ability of the cell metastasis and invasion.Results The expression of E-cadherin was decreased with the increase of A1 AT concentration,and the ability of cell metastasis and invasion was better with the increasing expression of N-cadherin and Vimentin.Conclusion Exogenous A1 AT promotes the metastasis and invasion of non-small cell lung cancer.
Objective To investigate the effect of alpha 1-antitrypsin(A1 AT)on the promotion of metastasis and invasion of non-small cell lung cancer(NSCLC).Methods The different concentrations of A1 AT(0,600,1200,2400 ng/ml)were added into the cell culture solution.Western blot and immunofluorescence were used to detect the epithelial cell marker E-cadherin and the mesenchymal markers including N-cadherin and Vimentin.The transwell and wound-healing assays were used to detect the ability of the cell metastasis and invasion.Results The expression of E-cadherin was decreased with the increase of A1 AT concentration,and the ability of cell metastasis and invasion was better with the increasing expression of N-cadherin and Vimentin.Conclusion Exogenous A1 AT promotes the metastasis and invasion of non-small cell lung cancer.
引文
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[27]Zheng X,Song T,Dou C,et al.CtBP2is an independent prognostic marker that promotes GLI1induced epithelialmesenchymal transition in hepatocellular carcinoma[J].Oncotarget,2015,6(6):3752-3769.
[28]Zhang X L,Huang C X,Zhang J,et al.CtBP1is involved in epithelial-mesenchymal transition and is a potential therapeutic target for hepatocellular carcinoma[J].Oncol Rep,2013,30(2):809-814.
[29]Kalluri R,Neilson E G.Epithelial-mesenchymal transition and its implications for fibrosis[J].J Clin Invest,2003,112(12):1776-1784.
[30]Holz C,Niehr F,Boyko M,et al.Epithelial-mesenchymaltransition induced by EGFR activation interferes with cell migration and response to irradiation and cetuximab in head and neck cancer cells[J].Radiother Oncol,2011,101(1):158-164.
[2]Chen W,Zheng R,Baade P D,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[3]Custodio A,de Castro J.Strategies for maintenance therapy in advanced non-small cell lung cancer:current status,unanswered questions and future directions[J].Crit Rev Oncol Hematol,2012,82(3):338-360.
[4]Teckman J H,Blomenkamp K S.Pathophysiology of Alpha-1Antitrypsin Deficiency Liver Disease[J].Methods Mol Biol,2017,1639:1-8.
[5]Jaberie H,Naghibalhossaini F.Recombinant production of native humanα-1-antitrypsin protein in the liver HepG2cells[J].Biotechnol Lett,2016,38(10):1683-1690.
[6]Motawi T,Shaker O,Hussein R,et al.Polymorphisms ofα1-antitrypsin and Interleukin-6genes and the progression of hepatic cirrhosis in patients with a hepatitis C virus infection[J].Balkan J Med Gene,2016,19(2):35-44.
[7]Pervakova M Y,Emanuel V L,Titova O N,et al.The Diagnostic Value of Alpha-1-Antitrypsin Phenotype in Patients with Granulomatosis with Polyangiitis[J].Int J Rheumatol,2016,2016:1-5.
[8]Ni K,Umair Mukhtar Mian M,Meador C,et al.Oncostatin M and TNF-αInduce Alpha-1 Antitrypsin Production in Undifferentiated Adipose Stromal Cells[J].Stem Cells Dev,2017,26(20):1468-1476.
[9]Buggio M,Towe C,Annan A,et al.Pulmonary vasculature directed adenovirus increases epithelial lining fluid alpha-1antitrypsin levels[J].J Gene Med,2016,18(1/3):38-44.
[10]何小维,刘玉,罗志刚.α1抗胰蛋白酶及其临床应用[J].农产品加工(学刊),2006(5):14-16.
[11]Shakya R,Tarulli G A,Sheng L,et al.Mutant p53upregulates alpha-1 antitrypsin expression and promotes invasion in lung cancer[J].Oncogene,2017,36(31):4469-4480.
[12]Di Francesco A,Di Germanio C,Panda A C,et al.Novel RNA-binding activity of NQO1promotes SERPINA1mRNA translation[J].Free Radic Biol Med,2016,99:225-233.
[13]Frenzel E,Wrenger S,Immenschuh S,et al.Acute-phase proteinα1-antitrypsin--a novel regulator of angiopoietin-like protein 4transcription and secretion[J].J Immunol,2014,192(11):5354-5362.
[14]Wu D M,Zhang P,Liu R Y,et al.Phosphorylation and changes in the distribution of nucleolin promote tumor metastasis via the PI3K/Akt pathway in colorectal carcinoma[J].FEBS Lett,2014,588(10):1921-1929.
[15]Wang H,Shi J,Luo Y,et al.LIM and SH3protein 1induces TGFβ-mediated epithelial-mesenchymal transition in human colorectal cancer by regulating S100A4expression[J].Clin Cancer Res,2014,20(22):5835-5847.
[16]Zhou J,Wang J,Zeng Y,et al.Implication of epithelialmesenchymal transition in IGF1R-induced resistance to EGFR-TKIs in advanced non-small cell lung cancer[J].Oncotarget,2015,6(42):44332-44345.
[17]Torre L A,Bray F,Siegel R L,et al.Global cancer statistics,2012[J].CA Cancer J Clini,2015,65(2):87-108.
[18]de Serres F,Blanco I.Role of alpha-1antitrypsin in human health and disease[J].J Intern Med,2014,276(4):311-335.
[19]Janciauskiene S M,Bals R,Koczulla R,et al.The discovery ofα1-antitrypsin and its role in health and disease[J].Respir Med,2011,105(8):1129-1139.
[20]Carleo A,Chorostowskawynimko J,Koeck T,et al.Does urinary peptide content differ between COPD patients with and without inherited alpha-1antitrypsin deficiency[J].Int J Chron Obstruct Pulmon Dis,2017,12:829-837.
[21]El-Akawi Z J,Al-Hindawi F K,Bashir N A.Alpha-1antitrypsin(alpha1-AT)plasma levels in lung,prostate and breast cancer patients[J].Neuro Endocrinol Lett,2008,29(4):482-484.
[22]Pérez-Holanda S,Blanco I,Menéndez M,et al.Serum concentration of alpha-1antitrypsin is significantly higher in colorectal cancer patients than in healthy controls[J].BMC Cancer,2014,14:355.
[23]Nam S,Park T.Pathway-based evaluation in early onset colorectal cancer suggests focal adhesion and immunosuppression along with epithelial-mesenchymal transition[J].PLoS One,2012,7(4):e31685.
[24]Kalluri R,Weinberg R A.The basics of epithelialmesenchymal transition[J].J Clin Invest,2009,119(6):1420-1428.
[25]Zhang Q,Wang S Y,Nottke A C,et al.Redox sensor CtBP mediates hypoxia-induced tumor cell migration[J].Proc Natl Acad Sci USA,2006,103(24):9029-9033.
[26]Scheel C,Weinberg R A.Phenotypic plasticity and epithelialmesenchymal transitions in cancer and normal stem cells[J].Int J Cancer,2011,129(10):2310-2314.
[27]Zheng X,Song T,Dou C,et al.CtBP2is an independent prognostic marker that promotes GLI1induced epithelialmesenchymal transition in hepatocellular carcinoma[J].Oncotarget,2015,6(6):3752-3769.
[28]Zhang X L,Huang C X,Zhang J,et al.CtBP1is involved in epithelial-mesenchymal transition and is a potential therapeutic target for hepatocellular carcinoma[J].Oncol Rep,2013,30(2):809-814.
[29]Kalluri R,Neilson E G.Epithelial-mesenchymal transition and its implications for fibrosis[J].J Clin Invest,2003,112(12):1776-1784.
[30]Holz C,Niehr F,Boyko M,et al.Epithelial-mesenchymaltransition induced by EGFR activation interferes with cell migration and response to irradiation and cetuximab in head and neck cancer cells[J].Radiother Oncol,2011,101(1):158-164.