RNA干扰糖原合成激酶-3β对胰腺癌细胞生长的抑制作用  

Glycogen synthase kinase-3β RNA interference suppresses growth of pancreatic adenocarcinoma cells

作　　者：汪理[1] 刘涛[1] 勾善淼[1] 周伟[1] 王统玲[1] 陈立波[1] 王春友[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院普通外科,武汉430022

出　　处：《中华实验外科杂志》2012年第10期2032-2034,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年基金资助项目（30801098）

摘　　要：目的观察沉默糖原合成激酶-3p（GSK-3p）对人胰腺癌细胞株PANC-1增殖和存活的影响，探讨其分子机制。方法实验分组如下：阴性对照组为未转染的PANC一1细胞；载体对照组为稳定转染control shRNA的PANC-1细胞；实验组为稳定转染GSK-3β shRNA的PANC．1细胞。噻唑蓝（MTF）比色法连续7d检测各组细胞的吸光度（A）值，并绘制出细胞的生长曲线；流式细胞仪（FCM）检测各组细胞的凋亡和周期；电泳迁移率实验（EMSA）检测各组细胞中核因子（NF）．KB的DNA结合活性。结果转染GSK-3B短发卡RNA（shRNA）的实验组PANC．1细胞生长速度明显减慢；实验组细胞的早期凋亡比例为（5．38±0．33）％，较阴性对照组（1．71±0．08）％和载体对照组（1．68±0．11）％显著升高（P〈0．05）；实验组中G0／G1期细胞比例为（60．02±1．99）％，较阴性对照组（46．56±3．13）％和载体对照组（46．04±2．92）％显著升高（P〈0．05），发生明显的G，期阻滞；实验组细胞中代表NF-KB复合体数量的条带A值为2186．37±225．51，与阴性对照组（4005．39±203．64）和载体对照组（3793．32±310．34）比较显著减少（P〈0．05）；而上述对照组组间差异无统计学意义（P〉0．05）。结论靶向抑制GSK-3β可抑制胰腺癌细胞增殖，诱导其凋亡，其机制可能与下调NF-κB的转录活性有关。Objective To investigate the effects of glycogen synthase kinase-3β （GSK-3β） RNA interference on proliferation of pancreatic adenocarcinoma cells and to explore the molecular mechanism in- volved in the procedure. Methods PANC-1 cells stably expressing GSK-3β shRNA and control shRNA served as the experimental group and the vector control group respectively, and the cells that were not transfected as the negative control group. The growth curves of PANC-1 cells were designed by MT＇F assay. The apoptosis and cycle of cells were analyzed by using flow cytometry （FCM）. The nuclear factor-KB （NF-κB） DNA binding activity was detected by using electrophoretic mobility shift assay （EMSA） analysis of nuclear extracts. Results As compared with control groups, the growth capability of PANC-I cells in experimental group was decreased notably. In experimental group, the early apoptosis rate [ （5.38 ± 0. 33 ） % ] was significantly higher than the negative control group [ （ 1.71 ± 0. 08 ） % ] and the vector control group [ （ 1.68 ± 0. 11 ） % ] （ P 〈 0. 05 ）. In experimental group, the percentage of cells in Go and G1 phases [ （60. 02 ± 1.99） % ] was higher than in the negative control group [ （46. 56 ± 3. 13） % ] and the vector con- trol group [ （46. 04±2. 92）% ], while that in S phase cell was lower （P 〈0. 05）. Cell cycle was arrest at G1 phase. In experimental group, the band intensity index which was described as the NF-KB complex number was （2186. 37 ±225. 51 ）, which was lower than in the negative control group [ （4005.39 ± 203.64） ] and the vector control group [ （3793.32 ±310. 34） ], but there was no significant difference between the negative control group and the vector control group （ P 〉 0. 05 ）. Conclusion GSK-313 RNA interference may inhibit the proliferation and survival of pancreatic adenocarcinoma cells via NF-KB signal pathway.

关 键 词：胰腺癌 糖原合成激酶-3B 核因子-ΚB RNA干扰 

分 类 号：R735.9[医药卫生—肿瘤]