[关键词]
[摘要]
目的: 研究核转录因子STAT3的诱骗寡核苷酸(decoy ODNs)对人膀胱癌细胞系增殖的抑制作用,并探讨其作用机制。方法: 采用阳离子聚合物Sofast为转染试剂将STAT3 decoy ODNs转染入膀胱癌T24及5637细胞;用MTT法检测细胞增殖情况;流式细胞术检测ODNs的转染效率;荧光显微镜检测ODNs入核情况;RT-PCR及Western blot检测转染ODNs前后bcl-xl、cyclinD1和c-myc表达水平的变化。结果:ODNs可有效地转染至细胞内,部分可进入核内,转染效率呈剂量依赖关系;STAT3 decoy ODNs可抑制膀胱癌细胞的增殖,100 nmol/L组抑制率最明显,达40%~48%;100 nmol/L decoy ODNs明显抑制bcl-xl、cyclinD1和c-myc mRNA及蛋白的表达水平。结论:STAT3 decoy ODNs可显著抑制膀胱癌细胞的增殖,其作用机制可能是通过下调癌基因c-myc、细胞周期基因cyclinD1及凋亡相关基因bcl-xl的基因转录及蛋白表达而实现的。
[Key word]
[Abstract]
Objective: To investigate the inhibitory effects of STAT3 decoy ODNs on the growth of human bladder cancer cell lines in vitro. Methods: STAT3 decoy and scramble ODNs were transfected into the bladder cell lines (T24 cells and 5637 cells) with Sofast, a positive compound transfection agent. MTT assay was used to detect cell growth rate; the transfection efficiency was detected by flow cytometry assay;the locations of FITC labeled decoy ODNs were observed by reflected light fluorescence microscope;and the expression of STAT3 downstream genes, such as bcl-xl, cyclinD1 and dc-myc mRNA and proteins, were examined by RT-PCR and Western bloting. Results: STAT3 decoy ODNs were effectively incorporated into the bladder cancer cells in a dose-dependent manner, and they inhibited the proliferation of cancer cells. The most obvious inhibition rate (40 % to 48%) was found when the concentration of STAT3 decoy ODNs was at 100 nmol/L. The expression levels of Bcl-xl,c-myc and CylinD1 mRNA and protein were also significantly inhibited by 100 nmol/L STAT3 decoy ODNs. Conclusion: STAT3 decoy ODNs can obviously inhibit the proliferation of the bladder cancer cell lines T24 and 5637 , which may be associated with the inhibition of STAT3-mediated gene expression, such as bcl-xl,c-myc, and cylinD1.
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[基金项目]
国家重点基础研究发展规划(973)资助项目(2004CB 518807)和国家自然科学基金项目(30571696)