BUB1B在原发性肝癌中的表达及对肝癌细胞增殖和侵袭的影响
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摘要
目的探究纺锤体检测蛋白BUB1B在原发性肝癌(HCC)患者中的表达及临床意义,并研究其在体外对肝癌细胞增殖和侵袭的影响。方法收集GEO数据库中HCC患者(GSE121248)的临床以及基因表达信息,通过生物信息学分析筛选差异表达基因,设计针对BUB1B基因的小干扰RNA载体,通过沉默BUB1B基因表达验证其在肝癌细胞增殖以及侵袭中的影响。利用实时荧光定量PCR检测沉默效率,通过细胞增殖实验以及划痕实验分别检测该基因在肝癌细胞增殖以及侵袭中的作用。利用KM-plotter数据库分析BUB1B基因表达高低对HCC患者预后的影响。结果沉默序列1和2能够有效地沉默BUB1B基因的mRNA水平表达(t=8.41,P=0.001;t=10.43, P <0.001),沉默BUB1B表达的肝癌细胞系HepG2的增殖能力减弱(t=13.94,P <0.001),肝癌细胞8 h的侵袭能力同样减小(t=14.94,P <0.001)。高表达BUB1B基因的HCC患者(n=127)生存率差于低表达BUB1B基因的HCC患者(n=237)[HR=2.01(1.42～2.86),P=6.6×10~(-5)],进一步分析发现,在有肝炎病毒感染史的HCC患者(n=150)中,BUB1B基因表达高低对预后没有影响[HR=1.88(0.89～3.99), P=0.093],只有在未感染肝炎病毒的HCC患者(n=167)中可以作为预测预后的分子标志物[HR=2.84(1.75～4.61), P=1.1×10~(-5)]。结论BUB1B基因在HCC患者中高表达,促进肝癌细胞增殖以及侵袭,在未感染肝炎病毒的HCC患者中,可以作为预测预后的潜在分子标志物。
Objective To investigate the expression and clinical signi?cance of BUB1 B in patients with human liver cancer cells in vitro. Methods The clinical data and the information of gene expression were collected from GEO database(GSE121248). The differentially-expressing genes were screened and analyzed via BUB1 B gene on the proliferation and invasion of liver cancer cells was assessed by silencing the expression of BUB1 B gene. The knockdown efficiency was detected by quantitative real-time PCR. The proliferating and invasion abilities were evaluated by cell proliferation assay and wound healing test. The effect of the expression levels Results The silence sequences 1 and 2 could signi?cantly silence the expression of BUB1 B mRNA(t=8.41, P=0.001; t=10.43, P < 0.001). The proliferating ability of hepatoma cell line HepG2, which knocked down the expression of BUB1 B gene, was signi?cantly weakened(t=13.94, P < 0.001). The invasion ability was also reduced at 8 h after knocking down the expression of BUB1 B gene(t=14.94, P < 0.001). The survival rate of HCC patients with high expression of BUB1 B gene(n=127) was signi?cantly lower than that of their counterparts with low expression of BUB1 B gene(n = 237) [HR = 2.01(1.42 ～ 2.86), P = 6.6×10~(-5)]. Further analysis found that the expression of BUB1 B gene exerted no effect on the clinical prognosis of HCC patients with a medical history of hepatitis(n = 150) [HR = 1.88(0.89 ～ 3.99), P=0.093], whereas it acted as a molecular biomarker to predict the clinical prognosis of HCC patients without a medical history of hepatitis(n = 167) [HR = 2.84(1.75 ～ 4.61), P = 1.1×10~(-5)]. Conclusions The BUB1 B gene is highly expressed in HCC patients, which can promote the proliferation and invasion of liver cancer cells. BUB1 B gene is a potential molecular biomarker for predicting the clinical prognosis of HCC patients with no medical history of hepatitis.
Objective To investigate the expression and clinical signi?cance of BUB1 B in patients with human liver cancer cells in vitro. Methods The clinical data and the information of gene expression were collected from GEO database(GSE121248). The differentially-expressing genes were screened and analyzed via BUB1 B gene on the proliferation and invasion of liver cancer cells was assessed by silencing the expression of BUB1 B gene. The knockdown efficiency was detected by quantitative real-time PCR. The proliferating and invasion abilities were evaluated by cell proliferation assay and wound healing test. The effect of the expression levels Results The silence sequences 1 and 2 could signi?cantly silence the expression of BUB1 B mRNA(t=8.41, P=0.001; t=10.43, P < 0.001). The proliferating ability of hepatoma cell line HepG2, which knocked down the expression of BUB1 B gene, was signi?cantly weakened(t=13.94, P < 0.001). The invasion ability was also reduced at 8 h after knocking down the expression of BUB1 B gene(t=14.94, P < 0.001). The survival rate of HCC patients with high expression of BUB1 B gene(n=127) was signi?cantly lower than that of their counterparts with low expression of BUB1 B gene(n = 237) [HR = 2.01(1.42 ～ 2.86), P = 6.6×10~(-5)]. Further analysis found that the expression of BUB1 B gene exerted no effect on the clinical prognosis of HCC patients with a medical history of hepatitis(n = 150) [HR = 1.88(0.89 ～ 3.99), P=0.093], whereas it acted as a molecular biomarker to predict the clinical prognosis of HCC patients without a medical history of hepatitis(n = 167) [HR = 2.84(1.75 ～ 4.61), P = 1.1×10~(-5)]. Conclusions The BUB1 B gene is highly expressed in HCC patients, which can promote the proliferation and invasion of liver cancer cells. BUB1 B gene is a potential molecular biomarker for predicting the clinical prognosis of HCC patients with no medical history of hepatitis.
引文
[1]Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA,Jemal A.Global cancer statistics 2018:GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185countries. CA Cancer J Clin,2018,68(6):394-424.
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[9]Barrett T, Wilhite SE, Ledoux P, Evangelista C, Kim IF,Tomashevsky M, Marshall KA, Phillippy KH, Sherman PM,Holko M, Yefanov A, Lee H, Zhang N, Robertson CL, Serova N, Davis S, Soboleva A.NCBI GEO:archive for functional genomics data sets——update. Nucleic Acids Res,2013,41(Database issue):D991-D995.
[10]Lánczky A, Nagyá, Bottai G, Munkácsy G, SzabóA, Santarpia L, Gy?rffy B.miRpower:a web-tool to validate survivalassociated miRNAs utilizing expression data from 2178 breast cancer patients. Breast Cancer Res Treat,2016,160(3):439-446.
[11]Lersritwimanmaen P, Nimanong S. Hepatocellular carcinoma surveillance:benefit of serum alfa-fetoprotein in real-world practice. Euroasian J Hepatogastroenterol,2018,8(1):83-87.
[12]Inagaki Y, Tang W, Makuuchi M, Hasegawa K, Sugawara Y,Kokudo N. Clinical and molecular insights into the hepatocellular carcinoma tumour marker des-γ-carboxyprothrombin. Liver Int,2011,31(1):22-35.
[13]Lee JH, Lee SW.The Roles of carcinoembryonic antigen in liver metastasis and therapeutic approaches. Gastroenterol Res Pract,2017,2017:7521987.
[14]Ma B, Liu X, Yu Z.The effect of high intensity focused ultrasound on the treatment of liver cancer and patients’ immunity.Cancer Biomark,2018.[Epub ahead of print]
[15]Chen XZ, Zhang WK, Tang HB, Li XJ, Tian GH, Shang HC,Li YS. The ethanol supernatant extracts of liushenwan could alleviate nanodiethylnitrosamine-induced liver cancer in mice.Can J Gastroenterol Hepatol,2018,2018:6934809.
[16]Ispas S, So S, Toy M. Barriers to disease monitoring and liver cancer surveillance among patients with chronic hepatitis B in the United States. J Community Health,2018.[Epub ahead of print]
[17]Tellapuri S, Sutphin PD, Beg MS, Singal AG, Kalva SP.Staging systems of hepatocellular carcinoma:a review. Indian J Gastroenterol,2018,37(6):481-491.
[18]Tu T, Bühler S, Bartenschlager R.Chronic viral hepatitis and its association with liver cancer. Biol Chem,2017,398(8):817-837.
[19]Islami F, Dikshit R, Mallath MK, Jemal A.Primary liver cancer deaths and related years of life lost attributable to hepatitis B and C viruses in India. Cancer Epidemiol,2016,40:79-86.
[2]Llovet JM, Montal R, Sia D, Finn RS.Molecular therapies and precision medicine for hepatocellular carcinoma. Nat Rev Clin Oncol,2018,15(10):599-616.
[3]陈万青,郑荣寿,张思维,曾红梅,邹小农,赫捷.2013年中国恶性肿瘤发病和死亡分析.中国肿瘤,2017,26(1):1-7.
[4]薛才林,杨涛,古诚鑫,蒋小峰,钱世鹍,刘世明,杨辉.溴结构域蛋白4抑制剂JQ1促使肝癌细胞侵袭迁移的分子机制.新医学,2017,48(7):443-448.
[5]Pinto M, Vieira J, Ribeiro FR, Soares MJ, Henrique R,Oliveira J, Jerónimo C, Teixeira MR.Overexpression of the mitotic checkpoint genes BUB1 and BUBR1 is associated with genomic complexity in clear cell kidney carcinomas. Cell Oncol,2008,30(5):389-395.
[6]Baker Malureanu L, van Ree JH, Crespo-Diaz R, Reyes S, Seaburg L, Shapiro V, Behfar A, Terzic A, van de Sluis B, van Deursen JM.Increased expression of BubR1 protects against aneuploidy and cancer and extends healthy lifespan. Nat Cell Biol,2013,15(1):96-102.
[7]Ding Y, Hubert CG, Herman J, Corrin P, Toledo CM, SkuttKakaria K, Vazquez J, Basom R, Zhang B, Risler JK, Pollard SM, Nam DH, Delrow JJ, Zhu J, Lee J, DeLuca J, Olson JM,Paddison PJ.Cancer-specific requirement for BUB1B/BUBR1 in human brain tumor isolates and genetically transformed cells.Cancer Discov,2013,3(2):198-211.
[8]Abal M, Obrador-Hevia A, Janssen KP, Casadome L, Menendez M, Carpentier S, Barillot E, Wagner M, Ansorge W,Moeslein G, Fsihi H, Bezrookove V, Reventos J, Louvard D,Capella G, Robine S.APC inactivation associates with abnormal mitosis completion and concomitant BUB1B/MAD2L1 upregulation. Gastroenterology,2007,132(7):2448-2458.
[9]Barrett T, Wilhite SE, Ledoux P, Evangelista C, Kim IF,Tomashevsky M, Marshall KA, Phillippy KH, Sherman PM,Holko M, Yefanov A, Lee H, Zhang N, Robertson CL, Serova N, Davis S, Soboleva A.NCBI GEO:archive for functional genomics data sets——update. Nucleic Acids Res,2013,41(Database issue):D991-D995.
[10]Lánczky A, Nagyá, Bottai G, Munkácsy G, SzabóA, Santarpia L, Gy?rffy B.miRpower:a web-tool to validate survivalassociated miRNAs utilizing expression data from 2178 breast cancer patients. Breast Cancer Res Treat,2016,160(3):439-446.
[11]Lersritwimanmaen P, Nimanong S. Hepatocellular carcinoma surveillance:benefit of serum alfa-fetoprotein in real-world practice. Euroasian J Hepatogastroenterol,2018,8(1):83-87.
[12]Inagaki Y, Tang W, Makuuchi M, Hasegawa K, Sugawara Y,Kokudo N. Clinical and molecular insights into the hepatocellular carcinoma tumour marker des-γ-carboxyprothrombin. Liver Int,2011,31(1):22-35.
[13]Lee JH, Lee SW.The Roles of carcinoembryonic antigen in liver metastasis and therapeutic approaches. Gastroenterol Res Pract,2017,2017:7521987.
[14]Ma B, Liu X, Yu Z.The effect of high intensity focused ultrasound on the treatment of liver cancer and patients’ immunity.Cancer Biomark,2018.[Epub ahead of print]
[15]Chen XZ, Zhang WK, Tang HB, Li XJ, Tian GH, Shang HC,Li YS. The ethanol supernatant extracts of liushenwan could alleviate nanodiethylnitrosamine-induced liver cancer in mice.Can J Gastroenterol Hepatol,2018,2018:6934809.
[16]Ispas S, So S, Toy M. Barriers to disease monitoring and liver cancer surveillance among patients with chronic hepatitis B in the United States. J Community Health,2018.[Epub ahead of print]
[17]Tellapuri S, Sutphin PD, Beg MS, Singal AG, Kalva SP.Staging systems of hepatocellular carcinoma:a review. Indian J Gastroenterol,2018,37(6):481-491.
[18]Tu T, Bühler S, Bartenschlager R.Chronic viral hepatitis and its association with liver cancer. Biol Chem,2017,398(8):817-837.
[19]Islami F, Dikshit R, Mallath MK, Jemal A.Primary liver cancer deaths and related years of life lost attributable to hepatitis B and C viruses in India. Cancer Epidemiol,2016,40:79-86.