miR-152靶向Mirk/Dyrk1B调控人卵巢癌干细胞紫杉醇敏感性的研究
|
摘要
目的探讨miR-152对人卵巢癌干细胞球紫杉醇敏感性的影响及可能的机制。方法分离培养人卵巢癌干细胞球,构建miR-152的差异表达体系,用CCK-8法检测紫杉醇作用下卵巢癌干细胞存活率;用荧光素酶报告基因系统鉴定miR-152与Mirk/Dyrk1B的靶向调控关系;蛋白印迹检测改变miR-152表达水平对Mirk/Dyrk1B及耐药相关蛋白的影响。结果过表达miR-152或降低Mirk/Dyrk1B表达水平可增强人卵巢癌干细胞球对紫杉醇的敏感性。miR-152能够特异性结合Mirk/Dyrk1B 3’-UTR区域。miR-152与Mirk/Dyrk1B差异表达影响人卵巢癌干细胞球耐药相关蛋白的表达水平。结论 miR-152可能通过靶向调节Mirk/Dyrk1B影响人卵巢癌干细胞球对紫杉醇的敏感性。
Objective To investigate the influence of miR-152 on the sensitivity of human ovarian cancer stem cells to paclitaxel and to understand the underlying mechanism. Methods The ovarian cancer stem cell spheres were isolated from the OVCAR3 cell line. miR-152 mimics were transfected into the ovarian cancer stem cells by lipofectamine 2000 and the viability of these cells after treatment with paclitaxel was examined by the CCK-8 assay. The regulatory relationship between miR-152 and Mirk/Dyrk1 B was investigated by the luciferase reporter assay. The protein levels of Mirk/Dyrk1 B and other drug resistance-related proteins were analyzed by Western blotting. Results Alteration of miR-152 or Mirk/Dyrk1 B could influence the sensitivity of ovarian cancer stem cells to paclitaxel and regulate the expression of drug resistance-related proteins. miR-152 could bind to the 3'-UTR of Mirk/Dyrk1 B m RNA. Conclusion miR-152 regulates the sensitivity of human ovarian cancer stem cells to paclitaxel by targeting Mirk/Dyrk1 B.
Objective To investigate the influence of miR-152 on the sensitivity of human ovarian cancer stem cells to paclitaxel and to understand the underlying mechanism. Methods The ovarian cancer stem cell spheres were isolated from the OVCAR3 cell line. miR-152 mimics were transfected into the ovarian cancer stem cells by lipofectamine 2000 and the viability of these cells after treatment with paclitaxel was examined by the CCK-8 assay. The regulatory relationship between miR-152 and Mirk/Dyrk1 B was investigated by the luciferase reporter assay. The protein levels of Mirk/Dyrk1 B and other drug resistance-related proteins were analyzed by Western blotting. Results Alteration of miR-152 or Mirk/Dyrk1 B could influence the sensitivity of ovarian cancer stem cells to paclitaxel and regulate the expression of drug resistance-related proteins. miR-152 could bind to the 3'-UTR of Mirk/Dyrk1 B m RNA. Conclusion miR-152 regulates the sensitivity of human ovarian cancer stem cells to paclitaxel by targeting Mirk/Dyrk1 B.
引文
[1]LAPIDOT T,SIRARD C,VORMOOR J,et al.A cell initiating human a-cute myeloid leukaemia after transplantation into SCID mice[J].Nature,1994,367(6464):645-648.DOI:10.1038/367645a0.
[2]REYA T,MORRISON SJ,CLARKE MF,et al.Stem cells,cancer,and cancer stem cells[J].Nature,2001,414(6859):105-111.DOI:10.1038/35102167.
[3]ZHANG YJ,LIU XC,DU J,et al.mi R-152 regulates metastases of non-small cell lung cancer cells by targeting neuropilin-1[J].Int J Clin Exp Pathol,2015,8(11):14235-14240.
[4]ZHAI R,KAN X,WANG B,et al.mi R-152 suppresses gastric cancer cell proliferation and motility by targeting CD151[J].Tumour Biol,2014,35(11):11367-11373.DOI:10.1007/s13277-014-2471-2.
[5]THEODORE SC,DAVIS M,ZHAO F,et al.Micro RNA profiling of novel African American and Caucasian prostate cancer cell lines reveals a reciprocal regulatory relationship of mi R-152 and DNA methyltranferase1[J].Oncotarget,2014,5(11):3512-3525.DOI:10.18632/oncotarget.1953.
[6]HUANG H,HU M,LI P,et al.mi R-152 inhibits cell proliferation and colony formation of CD133(+)liver cancer stem cells by targeting KIT[J].Tumour Biol,2015,36(2):921-928.DOI:10.1007/s13277-014-2719-x.
[7]ZHOU X,ZHAO F,WANG ZN,et al.Altered expression of mi R-152and mi R-148a in ovarian cancer is related to cell proliferation[J].Oncol Rep,2012,27(2):447-454.DOI:10.3892/or.2011.1482.
[8]LI LW,XIAO HQ,MA R,et al.mi R-152 is involved in the proliferation and metastasis of ovarian cancer through repression of ERBB3[J].Int J Mol Med,2018,41(3):1529-1535.DOI:10.3892/ijmm.2017.3324.
[9]QIN W,XIE W,HE Q,et al.Micro RNA-152 inhibits ovarian cancer cell proliferation and migration and may infer improved outcomes in ovarian cancer through targeting FOXP1[J].Exp Ther Med,2018,15(2):1672-1679.DOI:10.3892/etm.2017.5529.
[10]HE J,YU JJ,XU Q,et al.ATG14 by EGR1-MIR152 sensitizes ovarian cancer cells to cisplatin-induced apoptosis by inhibiting cyto-protective autophagy[J].Autophagy,2015,11(2):373-384.DOI:10.1080/15548627.2015.1009781.
[11]LIU L,ZOU J,WANG Q,et al.Novel micro RNAs expression of patients with chemotherapy drug-resistant and chemotherapy-sensitive epithelial ovarian cancer[J].Tumour Biol,2014,35(8):7713-7717.DOI:10.1007/s13277-014-1970-5.
[12]REAM L,NORRIS L,SAADEH FA,et al.A novel serum microRNA panel to discriminate benign from malignant ovarian disease[J].Cancer Lett,2015,(28)356:628-636.DOI:10.1016/j.canlet.2014.10.010.
[13]LEDER S,WEBER Y,ALTAFAJ X,et al.Cloning and characterization of Dyrk1B,a novel member of the DYRK family of protein kinases[J].Biochem Biophys Res Commun,1999(2):474-479.DOI:10.1006/bbrc.1998.9967.
[14]BAGGA S,BRACHT J,HUNTER S,et al.Regulation by let-7 and lin-4 mi RNAs results in target m RNA degradation[J].Cell,2005,122(4):553-563.DOI:10.1016/j.cell.2005.07.031.
[15]XIANG Y,MA N,WANG D,et al.mi R-152 and mi R-185 co-contribute to ovarian cancer cells cisplatin sensitivity by targeting DNMT1 directly:a novel epigenetic therapy independent of decitabine[J].Oncogene,2014,33(3):378-386.DOI:10.1038/onc.2012.575.
[16]DENG X,EWTON DZ,LI S,et al.The kinase Mirk/Dyrk1B mediates cell survival in pancreatic ductal adenocarcinoma[J].Cancer Res,2006,66(8):4149-4158.DOI:10.1158/0008-5472.CAN-05-3089.
[17]GAO J,ZHENG Z,RAWAL B,et al.Mirk/Dyrk1B,a novel therapeutic target mediates cell survival in non-small cell lung cancer cells[J].Cancer Biol Ther,2009,8(17):1671-1679.DOI:10.4161/cbt.8.17.9322.
[18]KELLAND IR.Preclinical perspectives on platinum resistance[J].Drug,2000,59(4):1-8.DOI:10.2165/00003495-200059004-00001.
[19]DENG X,FRIEDMAN E.Mirk kinase inhibition blocks the in vivo growth of pancreatic cancer cells[J].Genes Cancer,2014,5(9/10):337-347.DOI:10.18632/genesandcancer.29.
[20]JING H,EILEEN F.Depleting Mirk/Dyrk1B kinase increases cisplatin toxicity in ovarian cancer cells[J].Genes Cancer,2010,1(8):803-811.DOI:10.1177/1947601910377644.
[21]JING H,HOLLY D,FRIEDMAN EA.Ovarian cancer cells,not normal cells,are damaged by Mirk/Dyrk1B/Dyrk1B kinase Inhibition[J].Int J Cancer,2013,132(10):2258-2269.DOI:10.1002/ijc.27917.
[22]TIBERIO P,CAVADINI E,ABOLAFIO G,et al.4-oxo-N-(4-hydroxyphenyl)retinamide:two independent ways to kill cancer cells[J].PLo S One,2010,5(10):e13362.DOI:10.1371/journal.pone.0013362.
[23]SINGH R,DHANYAMRAJU PK,LAUTH M.DYRK1B blocks canonical and promotes non-canonical Hedgehog signaling through activation of the m TOR/AKT pathway[J].Oncotarget,2017,8(1):833-845.DOI:10.18632/oncotarget.13662.
[2]REYA T,MORRISON SJ,CLARKE MF,et al.Stem cells,cancer,and cancer stem cells[J].Nature,2001,414(6859):105-111.DOI:10.1038/35102167.
[3]ZHANG YJ,LIU XC,DU J,et al.mi R-152 regulates metastases of non-small cell lung cancer cells by targeting neuropilin-1[J].Int J Clin Exp Pathol,2015,8(11):14235-14240.
[4]ZHAI R,KAN X,WANG B,et al.mi R-152 suppresses gastric cancer cell proliferation and motility by targeting CD151[J].Tumour Biol,2014,35(11):11367-11373.DOI:10.1007/s13277-014-2471-2.
[5]THEODORE SC,DAVIS M,ZHAO F,et al.Micro RNA profiling of novel African American and Caucasian prostate cancer cell lines reveals a reciprocal regulatory relationship of mi R-152 and DNA methyltranferase1[J].Oncotarget,2014,5(11):3512-3525.DOI:10.18632/oncotarget.1953.
[6]HUANG H,HU M,LI P,et al.mi R-152 inhibits cell proliferation and colony formation of CD133(+)liver cancer stem cells by targeting KIT[J].Tumour Biol,2015,36(2):921-928.DOI:10.1007/s13277-014-2719-x.
[7]ZHOU X,ZHAO F,WANG ZN,et al.Altered expression of mi R-152and mi R-148a in ovarian cancer is related to cell proliferation[J].Oncol Rep,2012,27(2):447-454.DOI:10.3892/or.2011.1482.
[8]LI LW,XIAO HQ,MA R,et al.mi R-152 is involved in the proliferation and metastasis of ovarian cancer through repression of ERBB3[J].Int J Mol Med,2018,41(3):1529-1535.DOI:10.3892/ijmm.2017.3324.
[9]QIN W,XIE W,HE Q,et al.Micro RNA-152 inhibits ovarian cancer cell proliferation and migration and may infer improved outcomes in ovarian cancer through targeting FOXP1[J].Exp Ther Med,2018,15(2):1672-1679.DOI:10.3892/etm.2017.5529.
[10]HE J,YU JJ,XU Q,et al.ATG14 by EGR1-MIR152 sensitizes ovarian cancer cells to cisplatin-induced apoptosis by inhibiting cyto-protective autophagy[J].Autophagy,2015,11(2):373-384.DOI:10.1080/15548627.2015.1009781.
[11]LIU L,ZOU J,WANG Q,et al.Novel micro RNAs expression of patients with chemotherapy drug-resistant and chemotherapy-sensitive epithelial ovarian cancer[J].Tumour Biol,2014,35(8):7713-7717.DOI:10.1007/s13277-014-1970-5.
[12]REAM L,NORRIS L,SAADEH FA,et al.A novel serum microRNA panel to discriminate benign from malignant ovarian disease[J].Cancer Lett,2015,(28)356:628-636.DOI:10.1016/j.canlet.2014.10.010.
[13]LEDER S,WEBER Y,ALTAFAJ X,et al.Cloning and characterization of Dyrk1B,a novel member of the DYRK family of protein kinases[J].Biochem Biophys Res Commun,1999(2):474-479.DOI:10.1006/bbrc.1998.9967.
[14]BAGGA S,BRACHT J,HUNTER S,et al.Regulation by let-7 and lin-4 mi RNAs results in target m RNA degradation[J].Cell,2005,122(4):553-563.DOI:10.1016/j.cell.2005.07.031.
[15]XIANG Y,MA N,WANG D,et al.mi R-152 and mi R-185 co-contribute to ovarian cancer cells cisplatin sensitivity by targeting DNMT1 directly:a novel epigenetic therapy independent of decitabine[J].Oncogene,2014,33(3):378-386.DOI:10.1038/onc.2012.575.
[16]DENG X,EWTON DZ,LI S,et al.The kinase Mirk/Dyrk1B mediates cell survival in pancreatic ductal adenocarcinoma[J].Cancer Res,2006,66(8):4149-4158.DOI:10.1158/0008-5472.CAN-05-3089.
[17]GAO J,ZHENG Z,RAWAL B,et al.Mirk/Dyrk1B,a novel therapeutic target mediates cell survival in non-small cell lung cancer cells[J].Cancer Biol Ther,2009,8(17):1671-1679.DOI:10.4161/cbt.8.17.9322.
[18]KELLAND IR.Preclinical perspectives on platinum resistance[J].Drug,2000,59(4):1-8.DOI:10.2165/00003495-200059004-00001.
[19]DENG X,FRIEDMAN E.Mirk kinase inhibition blocks the in vivo growth of pancreatic cancer cells[J].Genes Cancer,2014,5(9/10):337-347.DOI:10.18632/genesandcancer.29.
[20]JING H,EILEEN F.Depleting Mirk/Dyrk1B kinase increases cisplatin toxicity in ovarian cancer cells[J].Genes Cancer,2010,1(8):803-811.DOI:10.1177/1947601910377644.
[21]JING H,HOLLY D,FRIEDMAN EA.Ovarian cancer cells,not normal cells,are damaged by Mirk/Dyrk1B/Dyrk1B kinase Inhibition[J].Int J Cancer,2013,132(10):2258-2269.DOI:10.1002/ijc.27917.
[22]TIBERIO P,CAVADINI E,ABOLAFIO G,et al.4-oxo-N-(4-hydroxyphenyl)retinamide:two independent ways to kill cancer cells[J].PLo S One,2010,5(10):e13362.DOI:10.1371/journal.pone.0013362.
[23]SINGH R,DHANYAMRAJU PK,LAUTH M.DYRK1B blocks canonical and promotes non-canonical Hedgehog signaling through activation of the m TOR/AKT pathway[J].Oncotarget,2017,8(1):833-845.DOI:10.18632/oncotarget.13662.