MicroRNA-506靶向Caveolin-1抑制绒毛膜癌细胞增殖和侵袭的研究
|
摘要
[目的]探讨microRNA-506 (miR-506)在绒毛膜癌中对绒毛膜癌细胞的作用及其作用机制。[方法]通过qRT-PCR检测绒毛膜癌和正常早孕绒毛临床标本中miR-506及caveolin-1表达量及相关性;将绒毛膜癌细胞JEG3和JAR按照miR-506 mimic (mimic)、miR-506 inhibitor(Inhi)、miR-506 mimic negative control(NC)和空白对照组(Blank)分别进行转染,通过MTT实验、Transwell实验和流式细胞术检测miR-506对绒毛膜细胞作用;通过双荧光素酶报告实验、qRT-PCR和Western blot检测miR-506、caveolin-1及相关蛋白的表达变化。[结果]与正常早孕绒毛相比,miR-506在绒毛膜癌组中表达显著降低(P<0.05),并与caveolin-1表达呈负相关。Mi R-506过表达明显抑制绒毛膜癌细胞的增殖、侵袭并促进凋亡。双荧光素酶报告实验表明miR-506过表达显著抑制野生型荧光素酶活性,qRT-PCR和Western blot结果显示miR-506过表达显著抑制caveolin-1表达,进而抑制相关蛋白表达。[结论] MiR-506可能通过靶向caveolin-1抑制绒毛膜癌细胞的增殖和侵袭。
[Purpose] To investigate the effect of microRNA-506 (miR-506) on proliferation and invasion ability of choriocarcinoma and its mechanisms. [Methods] The expression of miR-506 and caveolin-1 (CAV-1) in choriocarcinoma and normal early pregnancy villi was measured by qRTPCR. Human choriocarcinoma JEG3 JAR cells were transfected with miR-506 mimic,miR-506 inhibitor or miR-506 mimic negative control (NC),respectively. The cell proliferation,invasive ability and apoptosis were detected by MTT,transwell,and flow cytometry,respectively. The interaction,variation of miR-506 and CAV-1,the expressions of related proteins were detected by dual luciferase report assay,q RT-PCR and Western blot methods. [Results] Compared with normal early pregnancy villi,expression of miR-506 in choriocarcinoma significantly decreased (P<0.01) and it was negatively correlated with the expression of CAV-1. Dual luciferase report assay showed that miR-506 significantly inhibited activity of wild type luciferase in JEG3 and JAR cells. Overexpression of miR-506 significantly inhibited proliferation,invasion of choriocarcinoma cells and induced the cell apoptosis,and also significantly inhibited expression of CAV-1 and related proteins.[Conclusion] Mi R-506 may inhibit proliferation and invasion of choriocarcinoma cells via targeting CAV-1.
[Purpose] To investigate the effect of microRNA-506 (miR-506) on proliferation and invasion ability of choriocarcinoma and its mechanisms. [Methods] The expression of miR-506 and caveolin-1 (CAV-1) in choriocarcinoma and normal early pregnancy villi was measured by qRTPCR. Human choriocarcinoma JEG3 JAR cells were transfected with miR-506 mimic,miR-506 inhibitor or miR-506 mimic negative control (NC),respectively. The cell proliferation,invasive ability and apoptosis were detected by MTT,transwell,and flow cytometry,respectively. The interaction,variation of miR-506 and CAV-1,the expressions of related proteins were detected by dual luciferase report assay,q RT-PCR and Western blot methods. [Results] Compared with normal early pregnancy villi,expression of miR-506 in choriocarcinoma significantly decreased (P<0.01) and it was negatively correlated with the expression of CAV-1. Dual luciferase report assay showed that miR-506 significantly inhibited activity of wild type luciferase in JEG3 and JAR cells. Overexpression of miR-506 significantly inhibited proliferation,invasion of choriocarcinoma cells and induced the cell apoptosis,and also significantly inhibited expression of CAV-1 and related proteins.[Conclusion] Mi R-506 may inhibit proliferation and invasion of choriocarcinoma cells via targeting CAV-1.
引文
[1]Froeling FEM,Seckl MJ.Gestational trophoblastic tumours:an update for 2014[J].Curr Oncol Rep,2014,16(11):408-410.
[2]Seckl MJ,Sebire NJ,Berkowitz RS.Gestational trophoblastic disease[J].Lancet,2010,376(9742):717-729.
[3]Xie LS,Zhao TC,Cai J,et al.Methotrexate induces DNAdamage and inhibits homologous recombination repair in choriocarcinoma cells[J].Onco Targets Ther,2016,9:7115-7122.
[4]May T,Goldstein DP,Berkowitz RS.Current chemotherapeutic management of patients with gestational trophoblastic neoplasia[J].Chemother Res Pract,2011,2011:806256.
[5]Shi D,Tan Z,Lu R,et al.MicroRNA-218 inhibits the proliferation of human choriocarcinoma JEG-3 cell line by targeting Fbxw8[J].Biochem Biophys Res Commun,2014,450(4):1241-1246.
[6]Lee Y,Ahn C,Han JJ,et al.The nuclear RNaseⅢDrosha initiates microRNA processing[J].Nature,2003,425(6956):415-419.
[7]Gregory RI,Chendrimada TP,Cooch N,et al.Human RISCcouples microRNA biogenesis and posttranscriptional gene silencing[J].Cell,2005,123(4):631-640.
[8]Cheng Y,Dong L,Zhang J,et al.Recent advances in microRNA detection[J].Analyst,2018,143(8):1758-1774.
[9]Wang S,Wang N,Zheng Y,et al.Caveolin-1:an oxidative stress-related target for cancer prevention[J].Oxid Med Cell Longev,2017,2017:7454031.
[10]Fridolfsson HN,Roth DM,Insel PA,et al.Regulation o intracellular signaling and function by caveolin[J]FASEB J,2014,28(9):3823-3831.
[11]Wang Z,Wang N,Liu P,et al.Caveolin-1,a stress-related oncotarget,in drug resistance[J].Oncotarget,2015,6(35):37135-37150.
[12]Burgermeister E,Liscovitch M,Roecken C,et al.Caveats of caveolin-1 in cancer progression[J].Cancer Lett,2008,268(2):187-201.
[13]Zhang JY,Sun Y,Role of microRNA-506 in different tumors[J].Chinese Journal of Clinical Oncology 2016,43(3):121-124.[张靖宜,孙燕.MicroRNA-506在不同肿瘤中作用的研究进展[J].中国肿瘤临床,2016,43(3):120-124.]
[14]Hoffner L,Surti U.The genetics of gestational trophoblastic disease:a rare complication of pregnancy[J].Cancer Genet,2012,205(3):63-77.
[15]Cheung ANY,Zhang HJ,Xue WC,et al.Pathogenesis of choriocarcinoma:clinical,genetic and stem cell perspectives[J].Future Oncol,2009,5(2):217-231.
[16]Lin S,Gregory RI.MicroRNA biogenesis pathways in cancer[J].Nat Rev Cancer,2015,15(6):321-333.
[17]Kong KL,Kwong DLW,Chan THM,et al.MicroRNA-375inhibits tumour growth and metastasis in oesophageal squamous cell carcinoma through repressing insulin-like growth factor 1 receptor[J].Gut,2012,61(1):33-42.
[18]Zhao Y,Liu H,Li Y,et al.The role of miR-506 in transformed 16HBE cells induced by anti-benzo a pyrenetrans-7,8-dihydrodiol-9,10-epoxide[J].Toxicol Lett,2011,205(3):320-326.
[19]Yu Z,Zhang Y,Gao N,et al.Overexpression of miR-506inhibits growth of osteosarcoma through Snail2[J].Am JTransl Res,2015,7(12):2716-2723.
[20]Yang D,Sun Y,Hu L,et al.Integrated analyses identify a master microrna regulatory network for the mesenchymal subtype in serous ovarian cancer[J].Cancer Cell,2013,23(2):186-199.
[21]Tanase CP,Dima S,Mihai M,et al.Caveolin-1 overexpression correlates with tumour progression markers in pancreatic ductal adenocarcinoma[J].J Mol Histol,2009,40(1):23-29.
[22]Tang Y,Zeng X,He F,et al.Caveolin-1 is related to invasion,survival,and poor prognosis in hepatocellular cancer[J].Med Oncol,2012,29(2):977-984.
[23]Shan T,Lu H,Ji H,et al.Loss of stromal caveolin-1 expression:a novel tumor microenvironment biomarker that can predict poor clinical outcomes for pancreatic cancer[J].PLoS One,2014,9(6):e97239.
[2]Seckl MJ,Sebire NJ,Berkowitz RS.Gestational trophoblastic disease[J].Lancet,2010,376(9742):717-729.
[3]Xie LS,Zhao TC,Cai J,et al.Methotrexate induces DNAdamage and inhibits homologous recombination repair in choriocarcinoma cells[J].Onco Targets Ther,2016,9:7115-7122.
[4]May T,Goldstein DP,Berkowitz RS.Current chemotherapeutic management of patients with gestational trophoblastic neoplasia[J].Chemother Res Pract,2011,2011:806256.
[5]Shi D,Tan Z,Lu R,et al.MicroRNA-218 inhibits the proliferation of human choriocarcinoma JEG-3 cell line by targeting Fbxw8[J].Biochem Biophys Res Commun,2014,450(4):1241-1246.
[6]Lee Y,Ahn C,Han JJ,et al.The nuclear RNaseⅢDrosha initiates microRNA processing[J].Nature,2003,425(6956):415-419.
[7]Gregory RI,Chendrimada TP,Cooch N,et al.Human RISCcouples microRNA biogenesis and posttranscriptional gene silencing[J].Cell,2005,123(4):631-640.
[8]Cheng Y,Dong L,Zhang J,et al.Recent advances in microRNA detection[J].Analyst,2018,143(8):1758-1774.
[9]Wang S,Wang N,Zheng Y,et al.Caveolin-1:an oxidative stress-related target for cancer prevention[J].Oxid Med Cell Longev,2017,2017:7454031.
[10]Fridolfsson HN,Roth DM,Insel PA,et al.Regulation o intracellular signaling and function by caveolin[J]FASEB J,2014,28(9):3823-3831.
[11]Wang Z,Wang N,Liu P,et al.Caveolin-1,a stress-related oncotarget,in drug resistance[J].Oncotarget,2015,6(35):37135-37150.
[12]Burgermeister E,Liscovitch M,Roecken C,et al.Caveats of caveolin-1 in cancer progression[J].Cancer Lett,2008,268(2):187-201.
[13]Zhang JY,Sun Y,Role of microRNA-506 in different tumors[J].Chinese Journal of Clinical Oncology 2016,43(3):121-124.[张靖宜,孙燕.MicroRNA-506在不同肿瘤中作用的研究进展[J].中国肿瘤临床,2016,43(3):120-124.]
[14]Hoffner L,Surti U.The genetics of gestational trophoblastic disease:a rare complication of pregnancy[J].Cancer Genet,2012,205(3):63-77.
[15]Cheung ANY,Zhang HJ,Xue WC,et al.Pathogenesis of choriocarcinoma:clinical,genetic and stem cell perspectives[J].Future Oncol,2009,5(2):217-231.
[16]Lin S,Gregory RI.MicroRNA biogenesis pathways in cancer[J].Nat Rev Cancer,2015,15(6):321-333.
[17]Kong KL,Kwong DLW,Chan THM,et al.MicroRNA-375inhibits tumour growth and metastasis in oesophageal squamous cell carcinoma through repressing insulin-like growth factor 1 receptor[J].Gut,2012,61(1):33-42.
[18]Zhao Y,Liu H,Li Y,et al.The role of miR-506 in transformed 16HBE cells induced by anti-benzo a pyrenetrans-7,8-dihydrodiol-9,10-epoxide[J].Toxicol Lett,2011,205(3):320-326.
[19]Yu Z,Zhang Y,Gao N,et al.Overexpression of miR-506inhibits growth of osteosarcoma through Snail2[J].Am JTransl Res,2015,7(12):2716-2723.
[20]Yang D,Sun Y,Hu L,et al.Integrated analyses identify a master microrna regulatory network for the mesenchymal subtype in serous ovarian cancer[J].Cancer Cell,2013,23(2):186-199.
[21]Tanase CP,Dima S,Mihai M,et al.Caveolin-1 overexpression correlates with tumour progression markers in pancreatic ductal adenocarcinoma[J].J Mol Histol,2009,40(1):23-29.
[22]Tang Y,Zeng X,He F,et al.Caveolin-1 is related to invasion,survival,and poor prognosis in hepatocellular cancer[J].Med Oncol,2012,29(2):977-984.
[23]Shan T,Lu H,Ji H,et al.Loss of stromal caveolin-1 expression:a novel tumor microenvironment biomarker that can predict poor clinical outcomes for pancreatic cancer[J].PLoS One,2014,9(6):e97239.