RNA干扰技术沉默STAT3对人肺癌细胞生长抑制的作用

RNA 干扰技术沉默 STAT3 对人肺癌细胞生长抑制的作用【摘要】 目的探讨 RNA 干扰技术沉默STAT3 基因表达对肺腺癌 A549 细胞的生长抑制作用。方法针对 STAT3mRNA 序列设计合成 2 对编码 siRNA 的 DNA 模板，构建pSUPER siRNA STAT3重组质粒，转染 A549细胞；采纳 RT PCR法检测重组质粒对STAT3 基因表达的影响；在荧光显微镜下观察细胞的凋亡情况。 结果成功构建pSUPER siRNA STAT3重组质粒，并成功转染 A549 细胞。特异性 siRNA 片段能有效降低 STAT3mRNA 水平，最大干扰效率达％，明显高于空质粒对比组；干扰作用于转染后 24h 即可出现，48h 达高峰，72h 降低。对应不同位点的两个 siRNA 片段对 STAT3 均可产生干扰作用，彼此间差别不大。在荧光显微镜下，与未转染细胞相比，转染siRNA 的 A549 细胞中凋亡细胞所占比例明显增加。结论 pSUPER siRNA STAT3可抑制STAT3 在人肺腺癌 A549 细胞中的表达，并抑制肿瘤细胞的生长，促进其凋亡。以STAT3 为靶点的 RNA 干扰技术可望成为肺癌基因治疗的新方法。【关键词】 肺癌；RNA 干扰；STAT3；凋亡 Abstract：ObjectiveTo study the effects of pSUPER siRNA STAT3 gene on the growth of A549 cells．MethodsTwo pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pSUPER siRNA STAT3．Which was transfected into A549 cells．The STAT3 expression in A549 were transfected with pSUPER siRNA STAT3，and it was detected by RT PCR and cells viability by fluorescence siRNA eukaryotic expression vector against STAT3 mRNA was successfully conctructed， which was transfected into A549 cells. The level of STAT3 mRNA in A549 cells was inhibited by the specific siRNAs．The decrease of STAT3 mRNA expression began to appear 24 hours after transfection．And the most apparent interfering efficiency was ％,48 hours after transfection，which was markedly higher than that in the cells transfected with the control siRNAs．Both siRNAs from different loci had interfering effect on STAT3 mRNA expres...