原钙黏蛋白10(PCDH10)抑制BXPC-3胰腺癌细胞的增殖以及侵袭和迁移  

Protocadherin 10(PCDH10) inhibits the proliferation,invasion and migration ability of BXPC-3 pancreatic cancer cells

作　　者：邱禅 卜小娜 胡丹[1] 姜政[1] 

机构地区：[1]重庆医科大学附属第一医院消化内科,重庆400016

出　　处：《细胞与分子免疫学杂志》2016年第2期163-167,172,共6页Chinese Journal of Cellular and Molecular Immunology

摘　　要：目的探讨原钙黏蛋白10(PCDH10)在胰腺癌细胞株中的表达情况以及生物学功能。方法利用反转录PCR检测PCDH10在CAPAN-1、PANC-1、ASPC-1、BXPC-3胰腺癌细胞以及HPDE6-C7人正常胰腺导管上皮细胞的表达情况。将质粒pc DNA3.1-PCDH10与质粒pc DNA3.1-vector通过脂质体转染至BXPC-3人胰腺癌细胞,转染后通过反转录PCR检测BXPC-3细胞中PCDH10 mRNA的水平、Western blot法检测其蛋白水平;CCK-8法和克隆形成实验检测细胞增殖情况,膜联蛋白Ⅴ-碘化丙啶双标记结合流式细胞术检测细胞凋亡,Transwell^(TM)侵袭和迁移实验、划痕实验检测BXPC-3细胞的侵袭和迁移。结果与正常胰腺导管上皮细胞相比较,PCDH10在CAPAN-1、PANC-1、BXPC-3胰腺癌细胞中表达明显下调。反转录PCR及Western blot法检测结果表明,转染pc DNA3.1-PCDH10的BXPC-3实验组细胞PCDH10表达明显高于转染pc DNA3.1-vector的对照组细胞。与对照组相比,过表达PCDH10后,BXPC-3细胞的增殖速度明显降低、克隆形成数明显减少;细胞凋亡率明显增加,细胞的侵袭、迁移能力明显降低。结论 PCDH10在胰腺癌细胞中表达下调,过表达PCDH10能够明显抑制BXPC-3胰腺癌细胞的增殖、侵袭和迁移能力,并且诱导BXPC-3细胞凋亡。Objective To explore the expression and biological function of protocadherin 10（ PCDH10） in pancreatic cancer cells. Methods Reverse transcription PCR（ RT-PCR） was used to detect the expression of PCDH10 in CAPAN-1,PANC-1,ASPC-1,BXPC-3 pancreatic cancer cells and the HPDE6-C7 normal human pancreatic ductal epithelial cells.Recombinant plasmid pc DNA3. 1-PCDH10 and empty vector pc DNA3. 1 were transfected into BXPC-3 pancreatic cancer cells via Lipofectamine～（TM）2000. After transfection,the mRNA expression of PCDH10 was examined by RT-PCR,and the protein level was detected by Western blotting. CCK-8 and colony formation assays were used to analyze the cell proliferation. Cell apoptosis was tested by flow cytometry combined with annexin V-FITC/PI staining. Transwell^TMand wound healing assays were performed to measure the invasion and migration ability of the cells. Results Compared with the normal pancreatic ductal epithelial cells,the expression of PCDH10 was obviously lower in the CAPAN-1,PANC-1,BXPC-3 cells. RT-PCR and Western blotting revealed that PCDH10 expression significantly increased in BXPC-3 cells transfected with plasmid pc DNA3. 1-PCDH10 compared with the ones with empty vector pc DNA3. 1. CCK-8 and colony formation assays showed that the PCDH10-transfected cells grew more slowly than the empty vector-transfected cells. Annexin V-FITC/PI staining combined with flow cytometry proved that the apoptosis in the PCDH10-transfected cells remarkably increased compared with that in the control group. A reduction of the invasion and migration ability was found obviously in the PCDH10-transfected cells by Transwell^TMassay. The wound healing assay also showed that the PCDH10-transfected cells spread the more slowly than the empty vector-transfected cells. Conclusion The expression of PCDH10 was down-regulated in the pancreatic cancer cells.PCDH10 over-expression could significantly induce cell apoptosis,and restrain proliferation,invasion and migration ability of BXPC-3 pancreatic cancer cells.

关 键 词：PCDH10 胰腺癌 转染 增殖 凋亡 侵袭 迁移 

分 类 号：R735.9[医药卫生—肿瘤] Q279[医药卫生—临床医学]