摘要
背景与目的:目前研究表明,JAK-STAT3信号通路的异常激活与细胞恶性转化密切相关。本实验观察JAK激酶抑制剂AG490对鼻咽癌CNE-2Z细胞增殖、凋亡及凋亡相关基因Survivin、Mcl-1表达的影响,探讨JAK-STAT3信号通路在CNE-2Z细胞凋亡调控中的作用。方法:AG490作用于体外培养的CNE-2Z细胞。MTT法检测细胞增殖,Hoechst33342荧光活染、流式细胞术检测细胞凋亡,逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测STAT3、Survivin和Mcl-1 mRNA表达,Western blot检测STAT3、磷酸化STAT3(p-STAT3)、Survivin和Mcl-1蛋白表达。结果:100μmol/LAG490作用24、48h后对CNE-2Z细胞增殖抑制率分别为37.95%和52.99%。流式细胞术显示,经50μmol/L和100μmol/LAG490作用24h后,细胞凋亡率由处理前的1.37%分别提高到9.30%和9.00%(P<0.01)。荧光显微镜下可见到典型的细胞凋亡形态学改变。细胞STAT3活化程度降低,Survivin、Mcl-1基因表达下调。结论:阻断JAK-STAT3信号通路可抑制CNE-2Z细胞增殖,并可能通过下调凋亡抑制基因Survivin、Mcl-1表达诱导细胞凋亡。
Background and Objective. Abnormal activation of Janus kinas/ signal transducer and activator of transcription 3 （JAK-STAT3） signaling pathway is closely related to malignant transformation of cells. This study was to investigate the effects of a JAK inhibitor, AG490, on the proliferation, apoptosis, and expressions of apoptosis-related survivin and Mcl-1 genes, and to explore the role of JAK-STAT3 signaling pathway in the regulation of cell apoptosis in nasopharyngeal carcinoma （NPC） cell line CNE-2Z. Methods： CNE-2Z cells were treated with different doses of AG490. The cell proliferation was measured using MTT array. Cell apoptosis was assessed by flow cytometry （FCM） and Hoechst33342 fluorescence staining. The mRNA levels of STAT3, survivin and Mcl-1 were detected by reverse transcriptionpolymerase chain reaction （RT-PCR）. The protein levels of STAT3, phosphoralated STAT3 （p-STAT3）, survivin and Mcl-1 were measured by Western blot. Results The proliferation inhibition rates of AG490 （100μmol/ L） on CNE-2Z cells after 24 h and 48 h treatment were 37.95% and 52.99%. After incubation with 50 μmol/L and 100μmol/L AG490 for 24 h, the apoptosis rates of CNE-2Z cells increased from 1.37% to 9.30% and 9.00%, respectively （P〈0.01）; typical changes in apoptotic morphology were observed by fluorescence microscopy; moreover, activation of STAT3 was inhibited, mRNA and protein levels of Mcl-1 and survivin were downregulated. Conclusion： Blocking of JAK-STAT3 signaling pathway in CNE-2Z cells could remarkably inhibit cell proliferation and inactivate STAT3, as well as induce cell apoptosis by the down-regulation of Mcl-1 and survivin.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2009年第1期33-38,共6页
Chinese Journal of Cancer
基金
广东省自然科学基金项目(No031963)~~
