组织激肽释放酶12对胰腺癌细胞增殖、迁移和侵袭的影响  

Effect of kallikrein 12 on the proliferation,invasion and migration of pancreatic cancer cells

作　　者：胡丕波 刘蒙蒙 陈炯 Hu Pibo;Liu Mengmeng;Chen Jiong(Department of General Surgery,Huzhou Central Hospital,Huzhou 313000,China;Department of Pathology,Huzhou Central Hospital,Huzhou 313000,China;Department of General Surgery,First Affiliated Hospital of USTC(Anhui Provincial Hospital),Hefei 230001,China)

机构地区：[1]湖州市中心医院普外科,湖州313000 [2]湖州市中心医院病理科,湖州313000 [3]中国科学技术大学附属第一医院(安徽省立医院)普外科,合肥230001

出　　处：《中华胰腺病杂志》2022年第6期446-452,共7页Chinese Journal of Pancreatology

基　　金：国家自然科学基金(81071985);安徽省国际科技合作计划项目(10080703038)。

摘　　要：目的探讨组织激肽释放酶12(KLK12)对胰腺癌细胞增殖、侵袭及迁移能力的影响。方法收集2014年2月至2018年10月间中国科学技术大学附属第一医院胆胰外科行根治性手术切除且病理确诊的95例胰腺癌及其对应的癌旁正常组织,采用免疫组织化学法检测胰腺癌组织KLK12的表达,分析其与胰腺癌临床病理特征的相关性。采用蛋白质免疫印迹法和荧光定量PCR法检测胰腺癌细胞株SW1990、PANC1及正常胰腺腺泡细胞株HPDE6-C7的KLK12蛋白和mRNA表达。通过质粒转染构建上调及下调KLK12表达的胰腺癌细胞株,以未转染的及转染携带阴性对照质粒的胰腺癌细胞作为空白对照组和阴性对照组,运用CCK8法和Transwell小室检测各组细胞增殖、侵袭及迁移能力。结果胰腺癌组织KLK12的阳性表达率显著高于癌旁组织(70.5%比29.5%,P<0.001),其表达与肿瘤分化程度低、TNM分期晚和淋巴结转移显著相关(P值均<0.05)。SW1990、PANC1细胞的KLK12蛋白及mRNA表达量均显著高于HPDE6-C7细胞(0.34±0.01、0.28±0.01比0.21±0.01,3.31±0.10、2.91±0.09比1.41±0.20,P值均<0.01)。下调KLKL12表达,培养72 h时,SW1990细胞增殖的A450值(0.94±0.02比1.16±0.05)、穿膜细胞数[(373.7±14.8)个比(726.0±11.8)个/高倍视野]、迁移细胞数[(696.0±13.1)个比(841.3±15.4)个/高倍视野]均显著下降;PANC1细胞的A450值(0.96±0.03比1.21±0.03)、穿膜细胞数[(556.3±13.6)个比(646.0±15.1)个/高倍视野]、迁移细胞数[(449.0±16.5)个比(595.7±8.6)个/高倍视野]也均显著下降。而上调KLK12表达,培养72 h时,SW1990细胞增殖的A450值(1.32±0.03比1.11±0.03)、穿膜细胞数[(556.3±22.2)比(402.7±10.5)个/高倍视野]、迁移细胞数[(639.3±16.5)比(433.0±11.8)个/高倍视野]均显著增加;PANC1细胞增殖的A450值(1.26±0.04比1.08±0.03)、穿膜细胞数[(571.0±17.4)个比(426.7±23.3)个/高倍视野]、迁移细胞数[(740.3±13.0)个比(442.7±10.3)个/高倍视野]均显�Objective To explore the effects of kallikrein 12(KLK12)on the proliferation,invasion and migration ability of pancreatic cancer cells.Methods Pancreatic cancer tissues and para-cancer tissues were collected from 95 patients who underwent radical surgical resection,and pathologically diagnosed as pancreatic cancer at the Department of Biliary and Pancreatic Surgery of the First Affiliated Hospital of USTC between February 2014 and October 2018.Expression of KLK12 in pancreatic cancer tissues was investigated using immunohistochemistry,and the correlation between KLK12 expression and clinicopathological characteristics of pancreatic cancer was analyzed.Western blot and qPCR methods were used to detect the expression of KLK12 protein and mRNA in pancreatic cancer cell line SW1990,PANC1 and normal pancreatic gland cells HPDE6-C7.Pancreatic cancer cell lines with up-regulated and down-regulated KLK12 expression were constructed by plasmid transfection.Non-transfected pancreatic cancer cells and transfected pancreatic cancer cells carrying negative control plasmids were used as blank control group and negative control group.CCK8 and Transwell chamber experiments were used to study the changes in cell proliferation,invasion and migration.Results The positive rate of KLK12 in pancreatic cancer tissues was significantly higher than that in para-cancer tissues(70.5%vs 29.5%,P<0.001),and was significantly related to low tumor differentiation,late TNM stage and lymph node metastasis(all P value<0.05).The expression of KLK12 protein and mRNA in SW1990 and PANC1 cell lines was higher than that in HPDE6-C7(0.34±0.01,0.28±0.01 vs 0.21±0.01;3.31±0.10,2.91±0.09 vs 1.41±0.20;all P value<0.01).In cells with down-regulated KLK12 expression and then cultured for 72 h,the A450 value of SW1990 cell proliferation(0.94±0.02 vs 1.16±0.05),the number of invading membrane penetrating cells(373.7±14.8 vs 726.0±11.8 per high magnification field)and the number of migrating penetrating cells(696.0±13.1 vs 841.3±15.4 per high magnifi

关 键 词：胰腺肿瘤 组织激肽释放酶类 细胞增殖 免疫组织化学 

分 类 号：R735.9[医药卫生—肿瘤]