沉默胰腺癌上调因子对胰腺癌细胞株吉西他滨敏感性的影响  

Silencing pancreatic adenocarcinoma up-regulated factor increases the sensitivity of pancreatic cancer cell lineto gemcitabine

作　　者：高崇崇[1] 徐小兰[2] 李非[1] 刘爽[1] 崔叶青[1] 孙海晨[1] 武玉多 

机构地区：[1]首都医科大学宣武医院普外科,100053 [2]中国科学院生物物理研究所

出　　处：《中华肝胆外科杂志》2017年第1期44-47,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金（81272756）

摘　　要：目的观察沉默胰腺癌上调因子（PAUF）对胰腺癌BxPC-3细胞吉西他滨敏感性的影响。方法BxPC-3细胞中PAUF基因呈高表达，分别应用PAUF-shCtrl、PAUF-shPAUF转染BxPC-3细胞得到PAUF高低表达不同的对照组和实验组。应用实时聚合酶链式反应（RT-PCR）及蛋白印迹（Western blot）检测进行沉默后验证。MTF比色法检测两株细胞在不同浓度吉西他滨（0、3．1、6．25、12．5、25、50、100、200nmol／L）作用下对细胞增殖抑制率的影响。流式细胞术检测不同浓度吉西他滨（0、75、100nmol／L）作用下对细胞凋亡率的影响。结果BxPC-3shCtrl细胞及亲本细胞PAUF mRNA的相对表达量为1．00±0．06、0．83±0．07，均显著高于BxPC-3shPAUF细胞的0．25±0．02（均P〈0．05）。BxPC-3shCtrl细胞及亲本细胞PAUF蛋白相对表达量为0．89±0．07、0．95±0．04，显著高于BxPC-3shPAUF细胞的0．31±0．03（均P〈0．05）。吉西他滨对BxPC-3shCtrl细胞的半数抑制浓度IC50为（22．88±2．43）nmol／L，显著高于BxPC-3shPAUF细胞的（1．06±0．02）nmol／L（P〈0．05）；BxPC-3shPAUF细胞的凋亡随着吉西他滨浓度的增加速度明显大于BxPC-3shCtrl细胞。结论PAUF沉默后明显增加胰腺癌BxPC-3细胞对吉西他滨的敏感性。Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene. Methods BxPC-3 cells, which overexpress PAUF, was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3shCtrl and BxPC-3shPAUF cells as control and experiment group. Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot, respectively. The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine （0, 3.1, 6.25, 12.5, 25, 50, 100, 200 nmol/L） were detected by MTT. Apoptosis rates in the cells treated with different concentrations of gemcitabine （0, 75, 100 nmol/L） were then observed by flow cytometry. Results The relative PAUF mRNA expression level in BxPC-3shCtrl and BxPC-3 cells were 1. 00±0.06 and 0.83± 0.07, which were significantly higher than that in BxPC-3 shPAUF cells （0.25 ± 0.02 ; both P 〈 0.05）. The relative PAUF protein expression level in BxPC-3shCtrl and BxPC-3 cells were 0.89 ± 0.07 and 0.95±0.04, which were significantly higher than that in BxPC-3 shPAUF cells （0.31 ±0.03 ; both P 〈 0.05 ）. The ICSO value of gemcitabine to Bx- PC-3shCtrl cell was （ 22.88 ± 2.43 ） nmol/L, which was significantly higher than that of BxPC-3shPAUF cells [ （ 1.06±0.02 ） nmol/L ; P 〈 0.051 ; apoptosis rate of BxPC-3 shPAUF cells treated by gemcitabine increased faster than that of BxPC-3shCtrl cells. Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine.

关 键 词：胰腺癌细胞株 胰腺癌上调因子 吉西他滨 药物敏感性 

分 类 号：R735.9[医药卫生—肿瘤]