沉默PTN表达抑制人胰腺细胞侵袭转移  

Research of the expression of pleiotrophin in human pancreatic cancer and its effect on invasion and metastasis

作　　者：符常波[1] 陆卫军[2] 刘志苏[1] 

机构地区：[1]武汉大学中南医院普外科,430071 [2]湖北省肿瘤医院肝胆胰科,武汉430079

出　　处：《中华实验外科杂志》2016年第1期16-18,共3页Chinese Journal of Experimental Surgery

摘　　要：目的观察多效应生长因子PTN在胰腺癌组织及癌旁组织中的表达水平，探讨PTN对胰腺癌细胞侵袭转移的影响。方法采用实时定量聚合酶链反应（Real-time PCR）及Western blot方法检测PTN在12对人胰腺癌组织标本中基因及蛋白水平表达。将PTN小干扰RNA（siRNA）转染至胰腺癌细胞MIAPaCa2、PANC-1中，通过Real—timePCR及Western blot检测沉默PTN后胰腺癌细胞PTN mRNA及蛋白表达以验证其干扰效果，Transwell检测沉默PTN表达后胰腺癌细胞侵袭及迁移能力的变化。细胞划痕实验检测其迁移能力的变化。结果PTN mRNA和蛋白水平在胰腺癌组织中均呈高表达，而在相应配对的癌旁组织中相对低表达。PTNmRNA在胰腺癌组织及癌旁组织中相对表达量分别为0．011±0．013、0．001±0．002。沉默PTN后其mRNA表达水平明显下降（0．049±0．012比0．121±0．015）。迁移试验显示Panc-1细胞跨膜数在空白组、阴性对照组及抑制组中分别为（178．1±5．2）、（171．3±11．1）、（58．2±6．7）个；侵袭实验显示Panc-1细胞跨膜数在3组中分别为（135．6±6．2）、（134．4±6．2）、（72．2±4．9）个，MIA PaCa2细胞跨膜数为（58．3±7．5）、（56．8±8．7）、（24．2±8．1）个。划痕实验显示MIA PaCa2细胞在3组中比例分别为0．41±0．03、0．42±0．11、0．73±0．03，而Panc-1细胞在3组中比例分别为0．54±0．02、0．57±0．03、0．82±0．02。结论PTN在胰腺癌组织中高表达，敲除PTN能抑制胰腺癌细胞的侵袭转移。Objective To research pleiotrophin （PTN） expression levels in pancreatic cancer tissues and paracancerous tissues and to investigate PTN impact on the metastasis of the pancreatic cancer cell invasion. Methods Using real - time quantitative polymerase chain reaction （ Real - time PCR） and Western blotting to detect PTN expression of the gene and protein level in 12 pairs of human pancreatic tissues. Using small interfering RNA （siRNA） against PTN to inhibit its expression in pancreatic cancer cell line MIA PaCa2 and PANC - 1. Real - time PCR and Western blotting detecting PTN mRNA and protein expression are used in order to verify its effectiveness. Transwell cell tests are used to detect changes of cell invasion and migration after PTN expression is knocked down in pancreatic cancer. Wound healing assays are used to detect changes in their migration ability. Results Expression levels of PTN mRNA and protein in pancreatic cancer tissues were way high,while matching the low table in the corresponding paracancerous tissues. The relative expression of PTN mRNA was respectable higher in cancer tissues than that in corresponding paracancerous tissues （0. 011 ±0. 013 vs. 0. 001 ±0. 002）. Knocking down PTN suppresses its expression at mRNA and protein levels significantly, which is testified by the test of Real - time PCR （0. 049 ±0. 012 vs. 0. 121 ±0. 015） as well as western blotting. The migration test shows that the number of cells of PANC - 1 crossing the membrane in blank group/negative group/suppression group is 178.1 ± 5.2, 171.3 ± 11.1,58.2 ±6. 7 respectively. The invasion test show that the number of cells of Panc - 1 crossing the membrane in blank group/negative group/suppression group is 135.6 ± 6. 2, 134. 4 ± 6. 2, 72. 2 ± 4. 9 respectively. The invasion test shows that the number of cells of MIA PaCa2 crossing the membrane in blank group/negative group/suppression group is 58.3 ± 7.5, 56. 8 ± 8. 7, 24. 2 ± 8. 1 respectively. The numbers of suppression group remain the lowest

关 键 词：胰腺癌 PTN 侵袭转移 

分 类 号：R735.2[医药卫生—肿瘤]