中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
24期
1889-1893
,共5页
刘军%厉天瑜%张宁%杨晓群%王志明%马骏%顾喜喜%范越%蔡定芳
劉軍%厲天瑜%張寧%楊曉群%王誌明%馬駿%顧喜喜%範越%蔡定芳
류군%려천유%장저%양효군%왕지명%마준%고희희%범월%채정방
胃肿瘤%白细胞介素1β%核因子-κB%微RNAs
胃腫瘤%白細胞介素1β%覈因子-κB%微RNAs
위종류%백세포개소1β%핵인자-κB%미RNAs
Gastric neoplasms%Interleukin-1 beta%NF-kappa B%MicroRNAs
目的 探讨白细胞介素1β(IL-1β)促进胃癌细胞生长的机制.方法 采用芯片筛选IL-1β处理后人胃癌细胞系(AGS)细胞中差异表达的miRNAs;应用生物信息学手段及双荧光素酶报告系统筛选并验证miR-425的靶基因;CCK-8检测细胞增殖率;流式细胞仪检测细胞的凋亡;实时聚合酶链反应(Real-time PCR)检测临床样本及细胞中miR-425的表达.结果 (1)IL-1β处理AGS细胞后,29个miRNAs表达上调,17个miRNAs表达下调.其中miR-425在IL-1β处理后表达上调最明显.Real-time PCR结果显示,相比癌旁组织,miR-425在胃癌组织中表达显著上调(均值1.41与5.81,P＜0.000 1);(2)使用生物信息学方法miRecords预测miR-425的潜在靶基因,经双荧光素酶报告检测,证实PTEN是miR-425的直接靶标;(3)使用核因子(NF)-κB单克隆抗体对AGS细胞行染色质免疫沉淀,证实NF-κB蛋白与miR-425的启动子中形成了复合物.荧光素酶报告基因的结果显示:miR-425启动子上B结合位点是IL-1β诱导miR-425转录激活必需的;(4)IL-1β诱导的miR-425表达上调促进细胞的增殖,抑制细胞凋亡.结论 IL-1β通过激活NF-κB诱导miR-425表达上调,负性调控PTEN进而促进胃癌细胞的增殖和抗凋亡作用.
目的 探討白細胞介素1β(IL-1β)促進胃癌細胞生長的機製.方法 採用芯片篩選IL-1β處理後人胃癌細胞繫(AGS)細胞中差異錶達的miRNAs;應用生物信息學手段及雙熒光素酶報告繫統篩選併驗證miR-425的靶基因;CCK-8檢測細胞增殖率;流式細胞儀檢測細胞的凋亡;實時聚閤酶鏈反應(Real-time PCR)檢測臨床樣本及細胞中miR-425的錶達.結果 (1)IL-1β處理AGS細胞後,29箇miRNAs錶達上調,17箇miRNAs錶達下調.其中miR-425在IL-1β處理後錶達上調最明顯.Real-time PCR結果顯示,相比癌徬組織,miR-425在胃癌組織中錶達顯著上調(均值1.41與5.81,P＜0.000 1);(2)使用生物信息學方法miRecords預測miR-425的潛在靶基因,經雙熒光素酶報告檢測,證實PTEN是miR-425的直接靶標;(3)使用覈因子(NF)-κB單剋隆抗體對AGS細胞行染色質免疫沉澱,證實NF-κB蛋白與miR-425的啟動子中形成瞭複閤物.熒光素酶報告基因的結果顯示:miR-425啟動子上B結閤位點是IL-1β誘導miR-425轉錄激活必需的;(4)IL-1β誘導的miR-425錶達上調促進細胞的增殖,抑製細胞凋亡.結論 IL-1β通過激活NF-κB誘導miR-425錶達上調,負性調控PTEN進而促進胃癌細胞的增殖和抗凋亡作用.
목적 탐토백세포개소1β(IL-1β)촉진위암세포생장적궤제.방법 채용심편사선IL-1β처리후인위암세포계(AGS)세포중차이표체적miRNAs;응용생물신식학수단급쌍형광소매보고계통사선병험증miR-425적파기인;CCK-8검측세포증식솔;류식세포의검측세포적조망;실시취합매련반응(Real-time PCR)검측림상양본급세포중miR-425적표체.결과 (1)IL-1β처리AGS세포후,29개miRNAs표체상조,17개miRNAs표체하조.기중miR-425재IL-1β처리후표체상조최명현.Real-time PCR결과현시,상비암방조직,miR-425재위암조직중표체현저상조(균치1.41여5.81,P＜0.000 1);(2)사용생물신식학방법miRecords예측miR-425적잠재파기인,경쌍형광소매보고검측,증실PTEN시miR-425적직접파표;(3)사용핵인자(NF)-κB단극륭항체대AGS세포행염색질면역침정,증실NF-κB단백여miR-425적계동자중형성료복합물.형광소매보고기인적결과현시:miR-425계동자상B결합위점시IL-1β유도miR-425전록격활필수적;(4)IL-1β유도적miR-425표체상조촉진세포적증식,억제세포조망.결론 IL-1β통과격활NF-κB유도miR-425표체상조,부성조공PTEN진이촉진위암세포적증식화항조망작용.
Objective To evaluate the mechanism of interleukin-1 (IL-1) in tumorigenesis.Methods The miRNA expression was profiled with the Exiqon miRCURYTM LNA Array System.And the miRecords algorithm and luciferase reporter assays were employed for identifying the targets of miR-425.Cell proliferation assay was performed with CCK8.Cell apoptosis assay was performed with flow cytometry.Results IL-1 β induced the up-regulation of miR-425,a negative expression regulator of phosphatase and tensin homolog (PTEN).An increase in miR-425 depended upon IL-1 β-induced NF-kappaB activation.Repression of PTEN by miR-425 promoted gastric cancer cell proliferation.Conclusion There is a critical role for NF-kappa B-dependent up-regulation of miR-425.And it represents a new pathway for the repression of phosphatase and tensin homolog activation and the promotion of cell survival upon IL-1 β induction.
