Omi／HtrA2短发夹siRNA对肺癌细胞凋亡的影响

Effect of RNA interference targeting Omi/HtrA2 gene on AS49 apoptosis

目的检测肺癌组织中促凋亡因子Omi／HtrA2的表达,观察Omi／HtrA2的短发夹状小干扰RNA(shRNA-Omi／HtrA2)载体对肺癌细胞生长的影响。方法采用免疫组织化学SP法检测50例非小细胞肺癌和10例正常肺组织中Omi／HtrA2的表达;将shRNA-Omi／HtrA2载体转染至人肺腺癌细胞株A549,用逆转录-聚合酶链反应(RT-PCR)和Western blot检测其对Omi／HtrA2转录和表达的下调作用,流式细胞术检测Omi／HtrA2下调后A549细胞凋亡的变化。结果Omi／HtrA2在非小细胞肺癌中的表达明显高于正常肺组织(P<0．05)。shRNA-Omi／HtrA2载体能显著抑制A549细胞Omi／HtrA2基因的表达(88％)。转基因组细胞经顺铂处理后凋亡率为(13．00±0．98)％,显著低于对照组的凋亡率(24．00±1．08)％(P<0．05)。结论Omi／HtrA2的高表达参与肺癌的发展,Omi／HtrA2短发夹siRNA可抑制A549细胞的凋亡。

Objective To investigate the expression of pro-apoptotic factor Omi/HtrA2 in lung cancer and to explore the effect of RNA interference targeting Omi/HtrA2 gent on A549 apoptosis. Methods The immunohistochemical system is used to detect the expression of Omi/HtrA2 in tissues from 50 cases lung cancer and 10 normal cases. The effects of shRNA-Omi/HtrA2 on its transcription and translation were analyzed by reverse transeriptase-polyrnerase chain reaction （RT-PCR） and Western blot. The sensitivity of A549 cells to cisplatin （DDP） after Omi/HtrA2 gene silencing was detected by flow cytometry. Results The expression of Omi/HtrA2 in lung cancer was higher than in normal tissues （P 〈 0.05）. Omi/HtrA2 expression was inhibited about 88 % （ P 〈 0.05） after shRNA-Omi/HtrA2 transfected. The apoptosis rate of A549 cells to DDP was （ 13.00 ± 0.98） %, while the control group was （24.00 ± 1.08） % （P〈0.05） .Condusion The high expression of Omi/HtrA2 is closely related to the development of lung cancer. The shRNA-Omi/HtrA2 could inhibit the apoptosis of A549 cells.