胰腺癌中Legumain的表达及其对胰腺癌细胞增殖和迁移的影响  

Expression of Legumain in pancreatic cancer and its effects on proliferation and migration of pancreatic cancer cells

作　　者：张波 王正峰[1,2] 朱骏 严俊[2,4] 陈浩斐[1] 张宝腾[1] 周文策 ZHANG Bo;WANG Zheng-feng;ZHU Jun;YAN Jun;CHEN Hao-fei;ZHANG Bao-teng;ZHOU Wen-ce(the First Clinical Medical College,Lanzhou University,Lanzhou 730000,China;Department of General Surgery,the First Hospital of Lanzhou University,Lanzhou 730000,China;Department of Pathology in Donggang District,the First Hospital of Lanzhou University,Lanzhou 730000,China;Gansu Province Key Laboratory of Biotherapy and Regenerative Medicine,the First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一临床医学院,兰州730000 [2]兰州大学第一医院普外科,兰州730000 [3]兰州大学第一医院东岗院区病理科,兰州730000 [4]兰州大学第一医院甘肃省生物治疗与再生医学重点实验室,兰州730000

出　　处：《临床与实验病理学杂志》2021年第4期402-407,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：甘肃省科技重点研发计划资助课题(17YF1FA128);兰州市人才创新创业项目资助课题(2017-RC-37)。

摘　　要：目的探讨Legumain(LGMN)在胰腺癌组织和细胞中的表达、意义及其下调对胰腺癌AsPC-1细胞增殖和迁移的影响。方法采用免疫组化SP法检测LGMN蛋白在67例胰腺癌组织及61例癌旁组织中的表达;分析LGMN表达与胰腺癌临床病理特征的关系。采用RT-PCR和Western blot法检测胰腺细胞系中LGMN的表达。通过慢病毒感染下调AsPC-1细胞中LGMN的表达,以未干预的细胞作为空白对照组,转染空白载体细胞为阴性对照组,LGMN敲低细胞为实验组;采用CCK-8法、Transwell实验、Western blot法检测下调LGMN对细胞增殖、迁移以及凋亡相关蛋白的影响。结果与癌旁组织相比,胰腺癌组织中LGMN高表达率更高,差异有统计学意义(P<0.01);LGMN表达与肿瘤分化程度、淋巴结转移相关(P<0.05),与患者年龄、性别、肿瘤大小等无关(P>0.05)。AsPC-1细胞中LGMN mRNA及蛋白表达量均显著高于胰腺正常上皮细胞HPNE,差异有统计学意义(P<0.01);转染慢病毒后,与对照组相比,实验组的吸光度值、细胞迁移数量、BCL-2蛋白表达、BCL-2/Bax的比值降低(P<0.05),Bax升高差异不明显(P>0.05)。结论LGMN在胰腺癌中呈高表达,体外敲低LGMN后能够抑制胰腺癌细胞的增殖和迁移能力,并通过BCL-2/Bax激活凋亡通路诱导细胞凋亡。Purpose To investigate the expression and significance of Legumain(LGMN)in pancreatic cancer tissues and pancreatic cells and the effect of LGMN down-regulation on proliferation and migration of pancreatic cancer AsPC-1 cells.Methods Immunohistochemical SP method was used to detect the expression of LGMN protein in 67 cases of pancreatic cancer tissues and 61 cases of para-cancerous tissues.The relationship between the expression of LGMN with clinicopathological parameters was analyzed.RT-PCR and Western blot were used to detect the expression of LGMN in pancreatic cells.The expression of LGMN in AsPC-1 cells was down-regulated by lentivirus infection.The cells without intervention were used as blank control group,those transfected with blank vector were used as negative control group,and those with LGMN knockdown were used as experimental group.CCK-8 assay,Transwell assay,and Western blot were used to detect the effect of LGMN down-regulation on cell proliferation,migration,and apoptosis-related proteins.Results Compared with para-cancerous tissues,the high expression rate of LGMN was higher in pancreatic cancer tissues,with statistically significant difference(P<0.01).LGMN expression was associated with the degree of tumor differentiation and lymph node metastasis(P<0.05),but not with age,gender,tumor size.(P>0.05).The expression levels of LGMN mRNA and protein in AsPC-1 cells were significantly higher than those in normal pancreatic epithelial cells HPNE,with statistically significant differences(P<0.01).After transfection with lentivirus,compared with the control group,the absorbance(OD)value,cell migration numbers,BCL-2 protein expression and BCL-2/Bax ratio in the experimental group were decreased(P<0.05),while the increase of Bax was not significantly different(P>0.05).Conclusion LGMN is highly expressed in pancreatic cancer,knockdown in vitro can inhibit the proliferation and migration of pancreatic cancer cells,and induce apoptosis through BCL-2/Bax activating apoptosis pathway.

关 键 词：胰腺肿瘤 LEGUMAIN 增殖 迁移 免疫组织化学 

分 类 号：R735.9[医药卫生—肿瘤]