【目的】探讨RNA干扰抑制内膜癌RL95-2细胞XIAP基因表达及对细胞凋亡的影响。【方法】设计并合成XIAP基因特异性siRNA,转染人内膜癌RL95-2细胞,Real time RT-PCR检测转染后XIAP mRNA的变化,Western Blot检测 XIAP蛋白的变化,MTT 及流式细胞仪法检测细胞增值和凋亡的变化。【结果】XIAP siRNA转染后,特异性转染组mRNA转录量的相对倍数值为0.04±0.06,相对蛋白表达量分别为0.590±0.178,较各对照组明显减少(P<0.05);特异性转染组细胞生长抑制率为(47.86±4.46)%,较对照组明显增高(P<0.05)。【结论】体外实验表明,合成的 siRNA能在mRNA水平及蛋白水平有效抑制人内膜癌RL95-2细胞内 XIAP基因的转录及表达,进而显著的促进内膜癌细胞凋亡,其促凋亡机制仍有待进一步研究。
[Obj ective]To explore the effect of RNA interference(RNAi)targeting XIAP gene on cell ap-optosis of human endometrial carcinoma.[Methods]Specific small interference RNA(siRNA)of XIAP was de-signed and composed.Endometrial cancer cell line RL95-2 was transfected.The changes of mRNA and protein of XIAP after transfection were detected by real time RT-PCR and Western Blotting,respectively.MTT and flow cytometry method were used to detect cell proliferation and apoptosis.[Results]After transfection of XI-AP siRNA,the relative fold value of mRNA transcription volume and the relative protein expression volume in specific transfection group were (0.04±0.06)and (0.590±0.178)respectively,which were obviously lower than those in the control group(P〈0.05).The cell growth inhibition rate in specific transduction group was (47.86±4.46)%,which was obviously higher than that in control group(P〈0.05).[Conclusion]The ex-periment in vitro indicates that the composed siRNA can effectively inhibit the transcription and expression of XIAP in human endometrial cancer cell line RL95-2,and further promote cell apoptosis of endometrial carcino-ma significantly.The mechanism of promoting cell apoptosis still needs further investigation.