胰腺癌曲古霉素A耐药细胞株转移相关基因表达的研究  

Enhanced Expression of Genes Associated with Invasion and Metastasis in Trichostatin A Resistance Pancreatic Cancer Cells Line

作　　者：刘彪[1] 陈孝倩[2] 张翠[2] 王本泉[1] 白永恒[2] 王斯璐[2] 金嵘[1] 周蒙滔[1] 陈必成[1,2] 

机构地区：[1]温州医学院附属第一医院肝胆胰外科,温州325000 [2]温州医学院附属第一医院外科实验室,温州325000

出　　处：《中国细胞生物学学报》2013年第8期1126-1132,共7页Chinese Journal of Cell Biology

基　　金：浙江省教育厅重中之重学科-外科学和温州市科技局项目(批准号:Y20090028)资助的课题~~

摘　　要：该实验旨在探讨曲古霉素A（trichostatin A,TSA）对胰腺癌PANC-1细胞株侵袭转移力的影响。分别通过短时间低浓度TSA（0.1~0.2μmol/L）处理胰腺癌细胞株PANC-1 24 h,以及长期低浓度递增法建立TSA耐药株（PANC-1-TSA）。采用CCK8（cell counting kit-8）测得PANC-1、PANC-1-TSA的IC50分别为（0.51±0.09）μmol/L、（78±5）μmol/L,耐药株耐药指数为153。Transwell侵袭小室实验显示,0.1μmol/LTSA处理后,胰腺癌PANC-1细胞侵袭力未见明显改变,而耐药株PANC-1-TSA侵袭力较亲本株明显增强。实时荧光定量PCR（RT-qPCR）结果显示,耐药株PANC-1-TSA转移相关基因MMP1/2/7/9/14/15及TIMP1/2 mRNA的表达均明显上升,尤以MMPs mRNA明显。在倒置显微镜观察到耐药细胞形态上发生上皮型向间叶型转化（epithelial mesenchymal transition,EMT）,进一步通过RT-qPCR及蛋白免疫印迹（Western blot）实验证实耐药株发生EMT,其中上皮型标志物表达下降及间叶型标志物表达上升。该实验显示,耐药细胞株PANC-1-TSA侵袭力增强,侵袭、转移相关基因表达明显增加。耐药株形成的过程中,发生上皮–间叶转化（EMT）,EMT可能参与了胰腺癌PANC-1-TSA细胞株侵袭能力的增强。The aim of this study was to investigate the change of invasion ability on PANC-1. PANC-1 cells line was treated with 0：1-0.2 μmol/L TSA for 24 hours. TSA-resistant pancreatic cancer cells line （PANC-1- TSA） was established by drug concentration step-elevation method. The 24 h IC50 of PANC-1 and PANC-1-TSA was （0.51±0.09） μmol/L and （78±5） μmol/L respectively, and the resistance index of PANC-1-TSA was 153. Tran- swell array experiment showed that PANC-1-TSA has stronger capability for invasion and metastasis than PANC-1 cells, while there was no significant difference in the 0.1 μmol/L TSA group. In RT-qPCR assessments, the expressions of MMPs and TIMPs mRNA were all up-regulated in PANC-1-TSA, and the MMPs mRNA increased much moresignificantly. We also found that PANC-1-TSA cells suffered morphological alteration from epithelial morphology to mesenchymal morphology. Both the RT-qPCR and Western blot experiments confirmed that the PANC-1 cells line was suffered EMT during the process of forming drug resistance, with up-regulation of mesenchymal markers and down- regulation of epithelial markers. This paper demonstrated that TSA-resistant cells line had stronger capability for invasion and metastasis and much more aggressive than PANC-1. EMT might contribute to PANC-1-TSA cells line aggression.

关 键 词：胰腺癌 古曲霉素A耐药 上皮–间叶转化 

分 类 号：R735.9[医药卫生—肿瘤]