Polo样激酶1对胰腺癌细胞株PANC-1凋亡、侵袭和迁移的影响  

Polo-like kinase 1 is involved in the apoptosis, invasion and metastasis of pancreatic cancer cell line PANC-1

作　　者：毛永欢 李泉[1] 蔡则灵 余翔[1] 喻春钊[1] Mao Yonghuan Li Quan Cai Zeling Yu Xiang Yu Chunzhao(Department of General Surgery, the Second Clinical Medical School of Nanjing Medical University, Nanjing 210029, China)

机构地区：[1]南京医科大学第二临床医学院普外科,210029

出　　处：《中华实验外科杂志》2016年第11期2535-2537,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（30972910、81172269）;中国博士后一等基金（20060390294）;江苏省自然科学基金（BK2011858）;江苏省六大人才高峰基金（WSW-032）

摘　　要：目的 观察Polo样激酶1（Plk1）基因对胰腺癌细胞株PANC-1凋亡、侵袭和迁移的影响.方法 实验分3组：对照组（Control）、空载组[rAd-增强型绿色荧光蛋白（EGFP）]、实验组[rAd-短发卡RNA（shRNA）].RNA干扰（RNAi）技术靶向沉默Plk1基因,构建人类Plk1-shRNA,通过腺病毒感染的方式导入体外培养的PANC-1细胞.实时定量反转录聚合酶链反应（RT-qPCR）和Western blot法检测Plk1在mRNA和蛋白水平的变化;流式细胞仪检测PANC-1细胞的凋亡变化;Transwell方法检测PANC-1细胞侵袭和迁移能力的改变.结果 RT-qPCR结果显示Plk1 mRNA水平表达量：对照组：1.011±0.153;空载组：0.943±0.052;实验组：0.256±0.008;Western blot结果显示Plk1蛋白水平表达量：对照组：0.920±0.029;空载组：0.883±0.029;实验组：0.013±0.005.实验组Plk1的表达量在mRNA和蛋白水平均明显低于对照组和空载组,差异有统计学意义（P＜0.01）.流式细胞术检测结果显示：下调Plk1,PANC-1细胞24 h后凋亡较其他两组明显增加（P＜0.01）;Transwell实验结果显示：侵袭能力,对照组：0.127±0.021;空载组：0.121±0.016;实验组：0.046±0.010;迁移能力,对照组：0.440±0.037;空载组：0.417±0.034;实验组：0.112±0.012;实验组细胞的侵袭和迁移能力较对照组和空载组明显下降,差异有统计学意义（P＜0.01）.结论 Plk1的表达与胰腺癌的凋亡、侵袭和转移密切相关,且采用RNAi技术靶向沉默Plk1基因可以促进胰腺癌细胞的凋亡,抑制其侵袭和迁移.Objective To investigate the effect of Polo-like kinase 1 （Plk1） gene on apoptosis,invasion and metastasis of pancreatic cancer cell PANC-1.Methods Human Plk1-short hairpin RNA （shRNA） were constructed to inhibit Plk1 gene by RNA interference （RNAi） technology,and PIk1-shRNA were transferred to PANC-1 cells by virus infection.Real-time reverse transcriptase-polymerase chain reaction （RT-qPCR） and Western blotting were used to detect the Plk1 mRNA and protein levels,and cell apoptosis was determined by flow cytometry.Transwell assay was used to examine the invasion and metastasis ability of PANC-1 cells.Results RT-qPCR showed Plk1 mRNA expression levels in control group,rAd-enhanced green fluorescent protein （EGFP） group and rAd-shRNA group were 1.011 ± 0.153,0.943 ± 0.052 and 0.256 ± 0.008 respectively （P 〈 0.01）.Western blotting showed Plk1 protein expression levels in control group,rAd-EGFP group and rAd-shRNA group were 0.920 ± 0.029,0.883 ± 0.029 and 0.013 ± 0.005 respectively （P 〈 0.01）.The flow cytometry indicated that the apoptosis was significantly increased in rAd-shRNA group after treatment for more than 24 h （P 〈 0.01）.Transwell assays revealed that the migratory and invasive capacities were significantly inhibited by infecting rAd-Plk1-shRNA （P 〈 0.01）.Conclusion Plk1 expression was closely correlated with the proliferation,invasion and metastasis of pancreatic cancer cells,and inhibition of Plk1 expression by RNAi can promote cell apoptosis and suppress metastasis and invasion of pancreatic cancer cells in vitro.

关 键 词：POLO样激酶1 胰腺癌 脱噬作用 侵袭 迁移 

分 类 号：R735.9[医药卫生—肿瘤]