特异性siRNA抑制XIAP基因表达对人内膜癌细胞凋亡的影响

Effect of Specific siRNA Inhibition of XIAP Expression on Cell Apoptosis of Human Endometri-al Carcinoma

【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.