沉默Yes相关蛋白1对人非小细胞肺癌细胞增殖和侵袭的影响
|
摘要
目的研究Yes相关蛋白1(YAP1)在非小细胞肺癌细胞增殖、细胞周期、迁移、侵袭中的作用。方法通过慢病毒介导的小干扰RNA靶向抑制人非小细胞肺癌细胞株A549中YAP1基因的表达,采用CCK-8法、流式细胞术分别检测沉默YAP1对A549细胞增殖及凋亡周期的影响,Transwell实验观察沉默YAP1对A549细胞迁移、侵袭能力的影响。结果沉默YAP1后,A549细胞YAP1mRNA和蛋白表达均下调(P<0.01),CCK-8实验结果显示沉默YAP1抑制A549的增殖、增加G0/G1期的细胞比例(P<0.01),Transwell实验结果示沉默YAP1抑制A549细胞的迁移、侵袭(P<0.01)。结论沉默YAP1基因对非小细胞肺癌细胞的恶性生物学特征具有抑制作用,YAP1可能成为肺癌治疗的潜在靶标。
Objective To explore the role of Yes-associated protein 1(YAP1)in the proliferation,cell cycle,migration and invasion of human non-small cell lung cancer(NSCLC)cells.Methods A lenti-siRNA targeting YAP1(si-YAP1)was used to inhibit the expression of YAP1 gene of human NSCLC cell line A549 cells.CCK-8assay and flow cytometry were used to determine the effects of silencing of YAP1 expression on A549 cells proliferation and cell cycle,respectively;Transwell assay was used to observe the effect of silencing of YAP1 expression on A549 cell migration and invasion.Results After infection with si-YAP1,the expressions of YAP1 mRNA and protein in A549 cells were significantly down-regulated(P<0.01).YAP1 silencing significantly inhibited A549 cell proliferation,increased the percentage of cells in G0/G1 phase(P<0.01),and significantly decreased the migration and invasion of A549cells(P<0.01).Conclusion YAP1 silencing can inhibit malignant biological characteristics of NSCLC,which suggests that YAP1 gene may serve as an important target in the gene therapy of lung cancer.
Objective To explore the role of Yes-associated protein 1(YAP1)in the proliferation,cell cycle,migration and invasion of human non-small cell lung cancer(NSCLC)cells.Methods A lenti-siRNA targeting YAP1(si-YAP1)was used to inhibit the expression of YAP1 gene of human NSCLC cell line A549 cells.CCK-8assay and flow cytometry were used to determine the effects of silencing of YAP1 expression on A549 cells proliferation and cell cycle,respectively;Transwell assay was used to observe the effect of silencing of YAP1 expression on A549 cell migration and invasion.Results After infection with si-YAP1,the expressions of YAP1 mRNA and protein in A549 cells were significantly down-regulated(P<0.01).YAP1 silencing significantly inhibited A549 cell proliferation,increased the percentage of cells in G0/G1 phase(P<0.01),and significantly decreased the migration and invasion of A549cells(P<0.01).Conclusion YAP1 silencing can inhibit malignant biological characteristics of NSCLC,which suggests that YAP1 gene may serve as an important target in the gene therapy of lung cancer.
引文
[1]JEMAL A,SIEGEL R,XU J,WARD E.Cancer statistics,2010[J].CA Cancer J Clin,2010,60:277-300.
[2]KODAKA M,HATA Y.The mammalian Hippo pathway:regulation and function of YAP1and TAZ[J].Cell Mol Life Sci,2015,72:285-306.
[3]YEUNG B,YU J,YANG X.Roles of the Hippo pathway in lung development and tumorigenesis[J].Int J Cancer,2016,138:533-539.
[4]ZANCONATO F,CORDENONSI M,PICCOLO S.YAP/TAZ at the roots of cancer[J].Cancer Cell,2016,29:783-803.
[5]XU M Z,YAO T J,LEE N P,NG I O,CHAN Y T,ZENDER L,et al.Yes-associated protein is an independent prognostic marker in hepatocellular carcinoma[J].Cancer,2009,115:4576-4585.
[6]LI M,LU J,ZHANG F,LI H,ZHANG B,WU X,et al.Yes-associated protein 1(YAP1)promotes human gallbladder tumor growth via activation of the AXL/MAPK pathway[J].Cancer Lett,2014,355:201-209.
[7]YU S J,HU J Y,KUANG X Y,LUO J M,HOU Y F,DI G H,et al.MicroRNA-200apromotes anoikis resistance and metastasis by targeting YAP1in human breast cancer[J].Clin Cancer Res,2013,19:1389-1399.
[8]NISHIKAWA E,OSADA H,OKAZAKI Y,ARIMA C,TOMIDA S,TATEMATSU Y,et al.miR-375is activated by ASH1and inhibits YAP1in a lineagedependent manner in lung cancer[J].Cancer Res,2011,71:6165-6173.
[9]KAPOOR A,YAO W,YING H,HUA S,LIEWEN A,WANG Q,et al.Yap1activation enables bypass of oncogenic Kras addiction in pancreatic cancer[J].Cell,2014,158:185-197.
[10]SONG S,AJANI J A,HONJO S,MARU D M,CHEN Q,SCOTT A W,et al.Hippo coactivator YAP1upregulates SOX9and endows esophageal cancer cells with stem-like properties[J].Cancer Res,2014,74:4170-4182.
[11]MURAMATSU T,IMOTO I,MATSUI T,KOZAKI K,HARUKI S,SUDOL M,et al.YAP is a candidate oncogene for esophageal squamous cell carcinoma[J].Carcinogenesis,2011,32:389-398.
[12]胡光辉,徐亮,赖鹏,郭锥锋,刘欢,刘敏,等.RNA干扰YAP基因对人膀胱癌T24细胞株增殖和迁移能力的影响[J].中国癌症杂志,2014,24:406-411.
[13]SUDOL M,BORK P,EINBONG A,KASTURY K,DRUCK T,NEGRINI M,et al.Characterization of the mammalian YAP(Yes-associated protein)gene and its role in defining a novel protein module,the WW domain[J].J Biol Chem,1995,270:14733-14741.
[14]TSCHAHARGANEH D F,CHEN X,LATZKO P,MALZ M,GAIDA M M,FELIX K,et al.Yesassociated protein up-regulates Jagged-1and activates the Notch pathway in human hepatocellular carcinoma[J].Gastroenterology,2013,144:1530-1542.
[15]SHAO D D,XUE W,KRALL E B,BHUTKAR A,PICCIONI F,WANG X,et al.KRAS and YAP1converge to regulate EMT and tumor survival[J].Cell,2014,158:171-184.
[16]YAMADA S,FUCHS B C,FUJII T,SHIMOYAMA Y,SUGIMOTO H,NOMOTO S,et al.Epithelial-tomesenchymal transition predicts prognosis of pancreatic cancer[J].Surgery,2013,154:946-954.
[2]KODAKA M,HATA Y.The mammalian Hippo pathway:regulation and function of YAP1and TAZ[J].Cell Mol Life Sci,2015,72:285-306.
[3]YEUNG B,YU J,YANG X.Roles of the Hippo pathway in lung development and tumorigenesis[J].Int J Cancer,2016,138:533-539.
[4]ZANCONATO F,CORDENONSI M,PICCOLO S.YAP/TAZ at the roots of cancer[J].Cancer Cell,2016,29:783-803.
[5]XU M Z,YAO T J,LEE N P,NG I O,CHAN Y T,ZENDER L,et al.Yes-associated protein is an independent prognostic marker in hepatocellular carcinoma[J].Cancer,2009,115:4576-4585.
[6]LI M,LU J,ZHANG F,LI H,ZHANG B,WU X,et al.Yes-associated protein 1(YAP1)promotes human gallbladder tumor growth via activation of the AXL/MAPK pathway[J].Cancer Lett,2014,355:201-209.
[7]YU S J,HU J Y,KUANG X Y,LUO J M,HOU Y F,DI G H,et al.MicroRNA-200apromotes anoikis resistance and metastasis by targeting YAP1in human breast cancer[J].Clin Cancer Res,2013,19:1389-1399.
[8]NISHIKAWA E,OSADA H,OKAZAKI Y,ARIMA C,TOMIDA S,TATEMATSU Y,et al.miR-375is activated by ASH1and inhibits YAP1in a lineagedependent manner in lung cancer[J].Cancer Res,2011,71:6165-6173.
[9]KAPOOR A,YAO W,YING H,HUA S,LIEWEN A,WANG Q,et al.Yap1activation enables bypass of oncogenic Kras addiction in pancreatic cancer[J].Cell,2014,158:185-197.
[10]SONG S,AJANI J A,HONJO S,MARU D M,CHEN Q,SCOTT A W,et al.Hippo coactivator YAP1upregulates SOX9and endows esophageal cancer cells with stem-like properties[J].Cancer Res,2014,74:4170-4182.
[11]MURAMATSU T,IMOTO I,MATSUI T,KOZAKI K,HARUKI S,SUDOL M,et al.YAP is a candidate oncogene for esophageal squamous cell carcinoma[J].Carcinogenesis,2011,32:389-398.
[12]胡光辉,徐亮,赖鹏,郭锥锋,刘欢,刘敏,等.RNA干扰YAP基因对人膀胱癌T24细胞株增殖和迁移能力的影响[J].中国癌症杂志,2014,24:406-411.
[13]SUDOL M,BORK P,EINBONG A,KASTURY K,DRUCK T,NEGRINI M,et al.Characterization of the mammalian YAP(Yes-associated protein)gene and its role in defining a novel protein module,the WW domain[J].J Biol Chem,1995,270:14733-14741.
[14]TSCHAHARGANEH D F,CHEN X,LATZKO P,MALZ M,GAIDA M M,FELIX K,et al.Yesassociated protein up-regulates Jagged-1and activates the Notch pathway in human hepatocellular carcinoma[J].Gastroenterology,2013,144:1530-1542.
[15]SHAO D D,XUE W,KRALL E B,BHUTKAR A,PICCIONI F,WANG X,et al.KRAS and YAP1converge to regulate EMT and tumor survival[J].Cell,2014,158:171-184.
[16]YAMADA S,FUCHS B C,FUJII T,SHIMOYAMA Y,SUGIMOTO H,NOMOTO S,et al.Epithelial-tomesenchymal transition predicts prognosis of pancreatic cancer[J].Surgery,2013,154:946-954.