CAJ | 학술논문

目的观察沉默糖原合成激酶-3β(GSK-3β)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-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(MTT)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)-κB的DNA结合活性.结果转染GSK-3β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G1期阻滞；实验组细胞中代表NF-κB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.
목적 관찰침묵당원합성격매-3β(GSK-3β)대인이선암세포주PANC-1증식화존활적영향,탐토기분자궤제.방법 실험분조여하:음성대조조위미전염적PANC-1세포；재체대조조위은정전염control shRNA적PANC-1세포；실험조위은정전염GSK-3β shRNA적PANC-1세포.새서람(MTT)비색법련속7d검측각조세포적흡광도(A)치,병회제출세포적생장곡선；류식세포의(FCM)검측각조세포적조망화주기；전영천이솔실험(EMSA)검측각조세포중핵인자(NF)-κB적DNA결합활성.결과 전염GSK-3β단발잡RNA (shRNA)적실험조PANC-1세포생장속도명현감만；실험조세포적조기조망비례위(5.38±0.33)％,교음성대조조(1.71±0.08)％화재체대조조(1.68 ±0.11)％현저승고(P＜0.05)；실험조중G0/G1기세포비례위(60.02±1.99)％,교음성대조조(46.56±3.13)％화재체대조조(46.04±2.92)％현저승고(P＜0.05),발생명현적G1기조체；실험조세포중대표NF-κB복합체수량적조대A치위2186.37±225.51,여음성대조조(4005.39±203.64)화재체대조조(3793.32 ±310.34)비교현저감소(P＜0.05)；이상술대조조조간차이무통계학의의(P＞0.05).결론 파향억제GSK-3β가억제이선암세포증식,유도기조망,기궤제가능여하조NF-κB적전록활성유관.
Objective To investigate the effects of glycogen synthase kinase-3 β (GSK-3β) RNA interference on proliferation of pancreatic adenocarcinoma cells and to explore the molecular mechanism involved 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 MTT assay.The apoptosis and cycle of cells were analyzed by using flow cytometry (FCM).The nuclear factor-κB (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-1 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 G0 and G1 phases [(60.02 ± 1.99)％] was higher than in the negative control group [(46.56 ±3.13)％] and the vector control 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-κB 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-3 β RNA interference may inhibit the proliferation and survival of pancreatic adenocarcinoma cells via NF-κB signal pathway.