敲低KCTD10对MHCC97H细胞体外致瘤性的影响
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摘要
目的:检定KCTD10敲低是否影响人肝细胞癌MHCC97H细胞体外致瘤能力和肿瘤干细胞相关标志物蛋白表达。方法:用表达KCTD10 Si RNA载体包装慢病毒感染,嘌呤霉素筛选构建KCTD10敲低MHCC97H细胞亚系。球形成法和平皿集落形成法比较慢病毒感染对照和KCTD10敲低MHCC97H细胞的体外致瘤能力。蛋白质印迹或流式细胞术分析肿瘤干细胞相关标志物蛋白表达。结果:敲低KCTD10能有效地增强球形成和平皿集落形成率。此外,敲低KCTD10上调ALDH1A1,BMI1和Nanog蛋白表达。结论:敲低KCTD10增强MHCC97H细胞体外致瘤能力和肿瘤干细胞样特性。
Objective To determine whether knock-down of KCTD10 affects in vitro tumorigenicity of human hepatocellular carcinoma(MHCC97 H) cells and the expression of cancer stem cell marker-related proteins. Methods KCTD10 knockdown MHCC97 H cell sublines was constructed by using lentiviral infection of packaged KCTD10 si RNA vector, puromycin screening. The capabilities of sphere formation and plate colony formation were compared between lentivirus infected control and KCTD10 knockdown Si RNA in MHCC97 H cells. Western blot or flow cytometry analysis were carried out for analyzing cancer stem cell-related marker protein expression. Results Knock-down of KCTD10 increased capabilities of the sphere formation and colony formation, and up-regulated cancer stem cell-related marker ALDH1 A1, BMI1 and Nanog protein expression. ConclusionKnock-down of KCTD10 enhances the tumorigenic in vitro and cancer stem cell characteristics of MHCC97 H cells.
Objective To determine whether knock-down of KCTD10 affects in vitro tumorigenicity of human hepatocellular carcinoma(MHCC97 H) cells and the expression of cancer stem cell marker-related proteins. Methods KCTD10 knockdown MHCC97 H cell sublines was constructed by using lentiviral infection of packaged KCTD10 si RNA vector, puromycin screening. The capabilities of sphere formation and plate colony formation were compared between lentivirus infected control and KCTD10 knockdown Si RNA in MHCC97 H cells. Western blot or flow cytometry analysis were carried out for analyzing cancer stem cell-related marker protein expression. Results Knock-down of KCTD10 increased capabilities of the sphere formation and colony formation, and up-regulated cancer stem cell-related marker ALDH1 A1, BMI1 and Nanog protein expression. ConclusionKnock-down of KCTD10 enhances the tumorigenic in vitro and cancer stem cell characteristics of MHCC97 H cells.
引文
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[2]Ghouri,Y.A.,I.Mian,et al.Review of hepatocellular carcinoma:Epidemiology,etiology,and carcinogenesis[J].J Carcinog,2017.16:1.
[3]Yan,X,Y.Qiu.Impact of current staging systems on treatment strategy for HBV-related hepatocellular carcinoma[J].Cancer Lett,2016.379(2):220-4.
[4]Collins,A.T..Prospective identification of tumorigenic prostate cancer stem cells[J].Cancer Res,2005.65(23):10946-51.
[5]Xu,X.L..The properties of tumor-initiating cells from a hepatocellular carcinoma patient's primary and recurrent tumor[J].Carcinogenesis,2010.31(2):167-74.
[6]Suetsugu,A.Characterization of CD133+hepatocellular carcinoma cells as cancer stem/progenitor cells[J].Biochem Biophys Res Commun,2006.351(4):820-4.
[7]Sun,J.K.[Preparation of mouse KCTD10 antibody and expression analysis of KCTD10 in neuroepithelium of neural tube and dorsal root ganglion][J].Sheng Wu Gong Cheng Xue Bao,2007.23(6):1011-6.
[8]Wang,Y.KCTD10 interacts with proliferating cell nuclear antigen and its down-regulation could inhibit cell proliferation[J].J Cell Biochem,2009.106(3):p.409-13.
[9]Kubota,D.Gene expression network analysis of ETV1 reveals KCTD10as a novel prognostic biomarker in gastrointestinal stromal tumor[J].PLoS One,2013.8(8):e73896.
[10]Maga,G.,U.Hubscher.Proliferating cell nuclear antigen(PCNA):a dancer with many partners[J].J Cell Sci,2003.116(Pt 15):3051-60.
[11]Zhou,J.A novel PDIP1-related protein,KCTD10,that interacts with proliferating cell nuclear antigen and DNA polymerase delta[J].Biochim Biophys Acta,2005.1729(3):200-3.
[12]Cao,L.Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines[J].BMC Gastroenterol,2011.11:71.
[13]Li,W.Gastric cancer-derived mesenchymal stem cells prompt gastric cancer progression through secretion of interleukin-8[J].J Exp Clin Cancer Res,2015.34:52.
[14]Jiang,F.Aldehyde dehydrogenase 1 is a tumor stem cell-associated marker in lung cancer[J].Mol Cancer Res,2009.7(3):330-8.
[15]Yanai,H.Intestinal cancer stem cells marked by Bmi1 or Lgr5 expression contribute to tumor propagation via clonal expansion[J].Sci Rep,2017.7:41838.
[16]Noh,K.H.Nanog signaling in cancer promotes stem-like phenotype and immune evasion[J].J Clin Invest,2012.122(11):4077-93.