泛素结合酶UBE2T在结直肠癌中的表达及其生物学作用
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摘要
目的探索泛素结合酶UBE2T在结直肠癌的表达及其生物学作用。方法运用免疫组化法检测78对结直肠癌和对应癌旁正常组织中UBE2T的蛋白表达水平。脂质体转染法将UBE2T-siRNA及对照NC-siRNA转染结直肠癌细胞SW480后,采用细胞计数试剂盒(CCK-8)及平板克隆实验分析增殖情况;流式细胞仪检测细胞周期情况;Western blotting检测细胞中UBE2T、cyclin D1、cyclin E等蛋白表达情况。结果与正常结直肠组织相比,结直肠癌组织中UBE2T表达水平增高(P<0.001)。癌组织中UBE2T异常高表达与不良病理指标相关:肿瘤大小(P=0.014)、组织分化程度(P=0.005)、浸润深度(P=0.033)和TNM分期(P=0.014)。CCK-8增殖实验和平板克隆形成实验都提示沉默UBE2T表达能显著抑制结直肠癌细胞的增殖。沉默UBE2T抑制周期蛋白cyclin D1和cyclin E的表达,促进结直肠癌细胞发生G1/S期细胞周期阻滞。结论 UBE2T在结直肠癌组织中异常高表达,并与不良病理指标相关。沉默UBE2T通过促进G1/S期细胞周期阻滞抑制结直肠癌细胞的增殖。
Objective To investigate the expression pattern and biological role of ubiquitin-binding enzyme UBE2 T in colorectal cancer.Methods Immunohistochemistry was used to detect the protein expression of UBE2 T in 78 pair specimens of colorectal cancer and corresponding normal tissues.After UBE2T-siRNA and control NC-siRNA were transfected into colorectal cancer cell SW480 by lipofectamine 2000,proliferation was evaluated by cell counting kit(CCK-8) and plate clone assay.Cell cycle was detected by flow cytometry.The protein expression of UBE2 T,cyclin D1 and cyclin E were detected by Western blotting.Results Compared with normal colorectal tissue,the expression of UBE2 T in colorectal cancer was significantly increased(P < 0.001).The abnormally high expression of UBE2 T in cancerous tissues was associated with poor pathological features:tumor size(P = 0.014),histological grade(P = 0.005),depth of invasion(P = 0.033) and TNM stage(P = 0.014).Both CCK-8 proliferation assay and plate clone formation assay suggested that silencing UBE2 T expression could significantly inhibit the proliferation of colorectal cancer cells.Silencing UBE2 T inhibits the expression of cyclin D1 and cyclin E,and promotes G1/S cell cycle arrest in colorectal cancer cells.Conclusion UBE2 T is highly expressed in colorectal cancer tissues and is associated with poor pathological parameters.Silencing UBE2 T inhibits the proliferation of colorectal cancer cells by promoting G1/S cell cycle arrest.
Objective To investigate the expression pattern and biological role of ubiquitin-binding enzyme UBE2 T in colorectal cancer.Methods Immunohistochemistry was used to detect the protein expression of UBE2 T in 78 pair specimens of colorectal cancer and corresponding normal tissues.After UBE2T-siRNA and control NC-siRNA were transfected into colorectal cancer cell SW480 by lipofectamine 2000,proliferation was evaluated by cell counting kit(CCK-8) and plate clone assay.Cell cycle was detected by flow cytometry.The protein expression of UBE2 T,cyclin D1 and cyclin E were detected by Western blotting.Results Compared with normal colorectal tissue,the expression of UBE2 T in colorectal cancer was significantly increased(P < 0.001).The abnormally high expression of UBE2 T in cancerous tissues was associated with poor pathological features:tumor size(P = 0.014),histological grade(P = 0.005),depth of invasion(P = 0.033) and TNM stage(P = 0.014).Both CCK-8 proliferation assay and plate clone formation assay suggested that silencing UBE2 T expression could significantly inhibit the proliferation of colorectal cancer cells.Silencing UBE2 T inhibits the expression of cyclin D1 and cyclin E,and promotes G1/S cell cycle arrest in colorectal cancer cells.Conclusion UBE2 T is highly expressed in colorectal cancer tissues and is associated with poor pathological parameters.Silencing UBE2 T inhibits the proliferation of colorectal cancer cells by promoting G1/S cell cycle arrest.
引文
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[16]POZNER A,TEROOATEA T W,BUCK-KOEHNTOP B A.Cell-specific kaiso(ZBTB33)regulation of cell cycle through cyclin D1 and cyclin E1[J].J Biol Chem,2016,291(47):24538-24550.
[2]LIU L P,YANG M,PENG Q Z,et al.UBE2T promotes hepatocellular carcinoma cell growth via ubiquitination of p53[J].Biochem Biophys Res Commun,2017,493(1):20-27.
[3]PEREZ-PEA J,CORRALES-SNCHEZ V,AMIR E,et al.Ubiquitin-conjugating enzyme E2T(UBE2T)and denticleless protein homolog(DTL)are linked to poor outcome in breast and lung cancers[J].Sci Rep,2017,7(1):17530.
[4]WEN M,KWON Y,WANG Y,et al.Elevated expression of UBE2T exhibits oncogenic properties in human prostate cancer[J].Oncotarget,2015,6(28):25226-25239.
[5]LUO C,YAO Y,YU Z,et al.UBE2T knockdown inhibits gastric cancer progression[J].Oncotarget,2017,8(20):32639-32654.
[6]李海青,曹猛,江涛,等.PD-L1在结直肠癌中的表达及其临床意义[J].徐州医学院学报,2017,37(4):227-230.
[7]李娟,尚颖,李丹,等.泛素化修饰在DNA损伤应答中的功能和作用机制研究进展[J].徐州医学院学报,2015,15(3):207-210.
[8]袁玉婷.泛素化修饰在盐敏感性高血压发生发展中的分子机制研究进展[J].陕西医学杂志,2017,46(8):1151-1152.
[9]BASSERMANN F,EICHNER R,PAGANO M.The ubiquitin proteasome system-implications for cell cycle control and the targeted treatment of cancer[J].Biochimi Biophys Acta,2014,1843(1):150-162.
[10]GALLO L H,KO J,DONOGHUE D J.The importance of regulatory ubiquitination in cancer and metastasis[J].Cell Cycle,2017,16(7):634-648.
[11]HIRA A,YOSHIDA K,SATO K,et al.Mutations in the gene encoding the E2 conjugating enzyme UBE2T cause fanconi anemia[J].Am J Hum Genet,2015,96(6):1001-1007.
[12]YU H,XIANG P,PAN Q,et al.Ubiquitin-conjugating enzyme E2T is an independent prognostic factor and promotes gastric cancer progression[J].Tumour Bio,2016,37(9):11723-11732.
[13]HU W,XIAO L,CAO C,et al.UBE2T promotes nasopharyngeal carcinoma cell proliferation,invasion,and metastasis by activating the AKT/GSK3beta/beta-catenin pathway[J].Oncotarget,2016,7(12):15161-15172.
[14]张玄,孙兴邦,赵晶,等.阿苯达唑对人胃癌细胞株MKN-45体外生长及细胞周期的影响[J].徐州医学院学报,2015,35(4):261-264.
[15]王兵.根治术后胃癌组织中肽基脯氨酰顺反异构酶1及细胞周期素E的表达及意义[J].中国基层医药,2013,20(10):1466-1468.
[16]POZNER A,TEROOATEA T W,BUCK-KOEHNTOP B A.Cell-specific kaiso(ZBTB33)regulation of cell cycle through cyclin D1 and cyclin E1[J].J Biol Chem,2016,291(47):24538-24550.