高迁移率族蛋白2在肝癌组织中的表达及对肝癌细胞生物学活性的影响
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摘要
目的:探究高迁移率族蛋白2(HMGB2)在肝细胞癌(HCC)组织中的表达及对HCC细胞生物学活性的影响。方法:利用TCGA和GEO公共数据库数据,分析HMGB2 mRNA在HCC组织中表达的特点。采用Western blot法检测正常肝细胞LO2和HCC细胞SMMC7721、Hep3B中HMGB2蛋白的表达。分别用HMGB2 siRNA和阴性对照siRNA转染SMMC7721和Hep3B细胞,采用CCK-8检测细胞增殖,Transwell体外迁移实验检测侵袭能力,流式法检测细胞周期和凋亡,以未转染细胞为空白对照。结果:HCC组织中HMGB2 mRNA的表达水平高于癌旁正常组织(P<0.05),TNM分期晚、分化程度低和甲胎蛋白高表达的HCC组织HMGB2 mRNA表达水平更高(P<0.05);Kaplan-Meier法生存分析结果显示,HMGB2 mRNA高表达者的总体生存期、无进展生存时间小于低表达者(P<0.05)。与空白对照和阴性对照siRNA转染细胞相比,HMGB2 siRNA转染的SMMC7721和Hep3B细胞中HMGB2蛋白表达降低,细胞增殖、侵袭能力降低(P<0.05),细胞被阻滞于G0/G1期(P<0.05),细胞凋亡率增加(P<0.05)。结论:HMGB2高表达能够促进HCC细胞的增殖、侵袭能力,有望成为HCC诊断及治疗的新靶点。
Aim:To investigate the expression of high mobility group box 2(HMGB2) in hepatocellular carcinoma(HCC) tissue and its effect on the biological activity of HCC cells.Methods:TCGA and GEO databases were used to analyze the expression character of HMGB2 mRNA in HCC tissue.The expression of HMGB2 protein in the cell lines LO2,SMMC7721 and Hep3 B was detected by Western blot.The biological function of SMMC7721 and Hep3 B cells transfected with HMGB2 siRNA or negative control siRNA was detected by CCK-8,Transwell assay,and flow cytometry.The cells without transfection were the blank control.Results:The expression of HMGB2 mRNA in HCC tissue was higher than that in adjacent normal tissue(P<0.05).High HMGB2 mRNA expression was significantly associated with late TNM stage,poor differentiation grade and high AFP level(P<0.05).Kaplan-Meier analysis showed that the overall survival and progression-free survival of patients with low HMGB2 mRNA expression were higher than those with high HMGB2 mRNA expression(P<0.05).Compared with the blank control and negative control siRNA group,the SMMC7721 and Hep3 B cells transfected with HMGB2 siRNA were inhibited in cell proliferation and invasion(P<0.05),arrested in G0/G1 phase(P<0.05) and with promoted cell apoptosis and lower expression of HMGB2(P<0.05).Conclusion:High HMGB2 expression could promote the proliferation and invasion abilities of HCC cells,and HMGB2 might be a potential target for HCC therapy.
Aim:To investigate the expression of high mobility group box 2(HMGB2) in hepatocellular carcinoma(HCC) tissue and its effect on the biological activity of HCC cells.Methods:TCGA and GEO databases were used to analyze the expression character of HMGB2 mRNA in HCC tissue.The expression of HMGB2 protein in the cell lines LO2,SMMC7721 and Hep3 B was detected by Western blot.The biological function of SMMC7721 and Hep3 B cells transfected with HMGB2 siRNA or negative control siRNA was detected by CCK-8,Transwell assay,and flow cytometry.The cells without transfection were the blank control.Results:The expression of HMGB2 mRNA in HCC tissue was higher than that in adjacent normal tissue(P<0.05).High HMGB2 mRNA expression was significantly associated with late TNM stage,poor differentiation grade and high AFP level(P<0.05).Kaplan-Meier analysis showed that the overall survival and progression-free survival of patients with low HMGB2 mRNA expression were higher than those with high HMGB2 mRNA expression(P<0.05).Compared with the blank control and negative control siRNA group,the SMMC7721 and Hep3 B cells transfected with HMGB2 siRNA were inhibited in cell proliferation and invasion(P<0.05),arrested in G0/G1 phase(P<0.05) and with promoted cell apoptosis and lower expression of HMGB2(P<0.05).Conclusion:High HMGB2 expression could promote the proliferation and invasion abilities of HCC cells,and HMGB2 might be a potential target for HCC therapy.
引文
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[20]邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70
[2]BRUIC J,SHERMAN M.Management of hepatocellular carcinoma:an update[J].Hepatology,2011,53(3):1020
[3]LI G,PU Y.MicroRNA signatures in total peripheral blood of gallbladder cancer patients[J].Tumor Biology,2015,36(9):6985
[4]PALLIER C,SCAFFIDI P,CHOPINEAUPROUST S,et al.Association of chromatin proteins high mobility group box(HMGB)1 and HMGB2 with mitotic chromosomes[J].Mol Biol Cell,2003,14(8):3414
[5]HOCK R,FURUSAWA T,UEDA T,et al.HMG chromosomal proteins in development and disease[J].Trends Cell Biol,2007,17(2):72
[6]杨如意,赵普学,许自强,等.人脑胶质瘤组织中HMGB1、TLR4的表达[J].郑州大学学报(医学版),2018,53(2):213
[7]ZHANG J,SHAO S,HAN D,et al.High mobility group box1 promotes the epithelial-to-mesenchymal transition in prostate cancer PC3 cells via the RAGE/NF-κB signaling pathway[J].Inter J Oncol,2018,53(2):659
[8]BORDAG M,SKALOZUB V.Altered expression and localization of creatine kinase B,heterogeneous nuclear ribonucleoprotein F,and high mobility group box 1 protein in the nuclear matrix associated with colon cancer[J].Cancer Res,2006,66(2):763
[9]HE Q,LIANG CH,LIPPARD SJ.Steroid hormones induce HMG1 overexpression and sensitize breast cancer cells to cisplatin and carboplatin[J].Proc Natl Acad Sci U S A,2000,97(11):5768
[10]LANGE SS,MITCHELL DL,VASQUEZ KM.High mobility group protein B1 enhances DNA repair and chromatin modification after DNA damage[J].Pro National Acad Sci USA,2008,105(30):10320
[11]LI JJ,GAO JH,TIAN W,et al.Long non-coding RNAMALAT1 drives gastric cancer progression by regulating HMGB2 modulating the miR-1297[J].Cancer Cell Int,2017,17(1):44
[12]REDMOND AM,BYRNE C,BANE FT,et al.Genomic interaction between ER and HMGB2 identifies DDX18 as a novel driver of endocrine resistance in breast cancer cells[J].Oncogene,2015,34(29):3871
[13]朱威威,陈晓龙,陈建安,等.α-烯醇化酶在肝癌组织中的表达及临床意义[J].郑州大学学报(医学版),2018,53(4):412
[14]张玥,曲春枫,任建松,等.中国肝癌发病与死亡数据集[J].中华肿瘤杂志,2015(9):705
[15]YAMANAKA Y,FAGHIHI MA,MAGISTRI MA,et al.Antisense RNA controls LRP1 sense transcript expression through interaction with a chromatin-associated protein,HMGB2[J].Cell Rep,2015,11(6):967
[16]TNIGUCHI N,CARAMS B,RONFANI L,et al.Aging-related loss of the chromatin protein HMGB2 in articular cartilage is linked to reduced cellularity and osteoarthritis[J].Proc Natl Acad Sci U S A,2009,106(4):1181
[17]WU Z,CAI L,LIN S,et al.High-mobility group box 2 is associated with prognosis of glioblastoma by promoting cell viability,invasion,and chemotherapeutic resistance[J].Neuro Oncol,2013,15(9):1264
[18]CAI X,DING HJ,LIU YX,et al.Expression of HMGB2 indicates worse survival of patients and is required for the maintenance of Warburg effect in pancreatic cancer[J].Acta Biochim Biophys Sin(Shanghai),2017,49(2):119
[19]KWON JH,KIM J,PARK JY,et al.Overexpression of highmobility group box 2 is associated with tumor aggressiveness and prognosis of hepatocellular carcinoma[J].Clin Cancer Res,2010,16(22):5511
[20]邹向阳,李连宏.细胞周期调控与肿瘤[J].国际遗传学杂志,2006,29(1):70