SUMO调控XBP1介导的内质网应激对肝癌进展的影响机制  

Mechanism of SUMO regulating XBP1 mediated endoplasmic reticulum stress on the progression of liver cancer

作　　者：辛瑞强 宋晓萍[2] 张帆 孙莹[2] 王涛[2] 孙伟 Xin Ruiqiang;Song Xiaoping;Zhang Fan;Sun Ying;Wang Tao;Sun Wei(Department of Hepatobiliary Surgery,Inner Mongolia People’s Hospital,Huhhot 010017,China;First Department of Respiratory and Critical Care,Affiliated Zhongshan Hospital of Dalian University,Dalian 116011,China;Department of Pathology,Inner Mongolia People’s Hospital,Huhhot 010017,China)

机构地区：[1]内蒙古自治区人民医院肝胆外科,呼和浩特010017 [2]大连大学附属中山医院呼吸与危重症一科,116011 [3]内蒙古自治区人民医院病理科,呼和浩特010017

出　　处：《国际肿瘤学杂志》2020年第7期397-403,共7页Journal of International Oncology

基　　金：大连市卫生健康委员会专项基金(1811114)。

摘　　要：目的探讨SUMO特异性蛋白酶1(SENP1)调控内质网应激转录调节因子X-盒结合蛋白1(XBP1)参与肝癌细胞增殖的分子机制。方法收集2012年1月至2020年1月内蒙古自治区人民医院肝胆外科收治的180例原发性肝癌患者的病理样本,检测肝癌、癌旁组织及不同肝癌细胞株中SENP1及XBP1的表达情况;分析肝癌组织中SENP1阳性表达与患者临床病理特征的相关性。免疫荧光与流式细胞技术检测SENP1 siRNA对XBP1及凋亡的影响。免疫沉淀检测肝癌细胞株中XBP1表面SUMO1表达情况及SENP1 siRNA对XBP1的SUMO化的影响。结果SENP1在肝癌及癌旁组织中的表达水平分别为16.332±4.371、6.840±2.238,差异具有统计学意义(t=-5.073,P=0.017)。XBP1在肝癌及癌旁组织中的表达水平分别为6.641±2.482、16.051±4.452,差异具有统计学意义(t=3.592,P=0.032)。肝癌组织中SENP1的表达与分期(χ^2=6.724,P=0.010)和是否转移(χ^2=6.265,P=0.012)相关。免疫荧光染色结果发现,XBP1表达水平在L02(0.509±0.219)、MHCC97-L(0.092±0.022)和HCCLM3(0.086±0.014)细胞间的表达差异有统计学意义(F=6.378,P=0.004),其中MHCC97-L和HCCLM3细胞显著低于在L02细胞的表达(P=0.023;P=0.021);SENP1在L02、MHCC97-L、HCCLM3细胞中的表达水平分别为0.109±0.079、0.802±0.392、0.921±0.352,差异具有统计学意义(F=7.783,P=0.004),其中MHCC97-L及HCCLM3细胞的表达均显著高于L02细胞(P=0.039;P=0.016);SENP1 siRNA转染MHCC97-L、HCCLM3细胞后,XBP1的表达水平均升高(0.462±0.192,t=3.664,P=0.022;0.524±0.203,t=3.383,P=0.028);SENP1的表达水平均下降(0.153±0.093,t=2.790,P=0.049;0.165±0.104,t=3.568,P=0.023)。流式细胞术结果显示,L02、MHCC97-L、HCCLM3、MHCC97-L+SENP1 siRNA及HCCLM3+SENP1 siRNA细胞的凋亡率分别为(20.80±3.11)%、(2.02±1.20)%、(0.12±0.01)%、(7.01±1.80)%、(6.20±2.01)%,差异具有统计学意义(F=1.025,P=0.030);MHCC97-L、HCCLM3细胞的凋亡率明显低于L02细胞(P=0.040;P=0.010);MHCC97-L+SENP1 siRNA�Objective To investigate the molecular mechanism of SUMO specific protease 1(SENP1)regulating endoplasmic reticulum stress transcription regulator X-box binding protein 1(XBP1)in the proliferation of liver cancer cells.Methods The pathological samples of 180 patients with primary liver cancer in the Department of Hepatobiliary Surgery of Inner Mongolia People's Hospital from January 2012 to January 2020 were collected.The expressions of SENP1 and XBP1 in liver cancer,adjacent tissues and different liver cancer cell lines were detected.The correlation between SENP1 positive expression and clinicopathological features of liver cancer patients was analyzed.Immunofluorescence and flow cytometry were used to detect the effect of SENP1 siRNA on XBP1 and apoptosis.SUMO1 expression on XBP1 surface was detected and the effect of SENP1 siRNA on SUMO formation of XBP1 was detected by immunoprecipitation.Results The expression levels of SENP1 in liver cancer and adjacent tissues were 16.332±4.371 and 6.840±2.238,with a statistically significant difference(t=-5.073,P=0.017).The expression levels of XBP1 in liver cancer and adjacent tissues were 6.641±2.482 and 16.051±4.452,with a statistically significant difference(t=3.592,P=0.032).The expression of SENP1 was correlated with stage(χ^2=6.724,P=0.010)and metastasis(χ^2=6.265,P=0.012).Immunofluorescence staining showed that the expressions of XBP1 in L02(0.509±0.219),MHCC97-L(0.092±0.022)and HCCLM3(0.086±0.014)cells were significantly different(F=6.378,P=0.004),while the expression of XBP1 in MHCC97-L and HCCLM3 cells was significantly lower than that in L02 cells(P=0.023;P=0.021).The expression levels of SENP1 in L02,MHCC97-L and HCCLM3 cells were 0.109±0.079,0.802±0.392 and 0.921±0.352,with a statistically significant difference(F=7.783,P=0.004),while the expression level of SENP1 in MHCC97-L and HCCLM3 cells was significantly higher than that in L02 cells(P=0.039;P=0.016).After transfection of SENP1 siRNA into MHCC97-L and HCCLM3 cells,the expressions of XBP1 inc

关 键 词：肝肿瘤 细胞凋亡 XBP1 SUMO 

分 类 号：R735.7[医药卫生—肿瘤]