Ad-IL-24抑制乳腺癌生长的体外实验研究

An experimental study on function of adenovirus-mediated IL-24 gene expression on suppressing breast cancer cells growth in vitro

目的研究腺病毒介导的IL-24基因(Ad-IL-24)表达对乳腺癌的生长抑制作用。方法将扩增的Ad-IL-24腺病毒感染人乳腺癌MDA-MB-231细胞,用RT-PCR法、Westernblot法检测IL-24基因在肿瘤细胞中的转录和表达;MTT法和FCM检测IL-24基因的表达对乳腺癌细胞的生长抑制和凋亡率;半定量RT-PCR检测IL-24基因的表达对乳腺癌细胞的凋亡相关基因Bax、Bcl-2及Survivin转录的影响;ELISA法检测VEGF、Ang-1的分泌水平。结果IL-24基因可在MDA-MB-231细胞中成功转录及表达,并对乳腺癌细胞增殖有明显抑制作用,并能引起细胞凋亡,其凋亡机制可能与Bax/Bcl-2比值升高、Survivin降低有关;VEGF、Ang-1表达下降。结论腺病毒介导的IL-24基因在体外可明显抑制人乳腺癌MDA-MB-231细胞的生长,诱导其凋亡,其机制可能与下调Survivin、VEGF、Ang-1,上调Bax的基因表达水平有关,该研究可为乳腺癌的基因治疗提供一定实验依据。

Objective To investigate the suppressing function of Ad-IL-24 on the proliferation of MDA-MB-231 cells.Methods Ad-IL-24 was obtained by virus-amplification technique.The expression of IL-24 gene in MDA-MB-231 breast cancer cells infected with Ad-IL-24 was detected by RT-PCR and Western blot;The growth suppressing and apoptosis effect were detected by MTT,flow cytometry respectively;RT-PCR was used to detect the transcription of Bax,Bcl-2,Survivin genes;The expression level of VEGF and Ang-1 gene were detected by ELISA.Results Il-24 was proved successfully transcribed and expressed in MDA-MB-231 cells and could obviously suppress proliferation of breast cancer cells and cause apoptosis of MDA-MB-231 cells,and its apoptosis mechanism might be related to increased ratio of Bax/Bcl-2 and decreased ratio of Survivin;The expression of VEGF and Ang-1 were down.Conclusion Ad-IL-24 could suppress the growth of MDA-MB-231 cells and induce their apoptosis,which might be related to gene levels of up-regulating Bax and down-regulating Survivin,VEGF、Ang-1.This study would provide certain experimental evidence for the gene therapy in breast cancer.