腺病毒介导黑色素瘤分化相关基因对乳腺癌细胞生长和凋亡的研究

Effects of adenoviral-mediated melanoma differentiation associated gene-7/IL-24 on growth and apoptosis of breast cancer cells

目的观察腺病毒介导黑色素瘤分化相关基因（mda-7／IL-24）对乳腺癌细胞生长和凋亡的作用。方法将腺病毒mda-7／IL-24进行病毒的扩增、纯化和滴度测定。将纯化好的腺病毒mda-7／IL-24感染乳腺癌细胞，用Western印迹方法检测mda-7／IL-24在乳腺癌细胞中的表达。结晶紫和4甲基偶氮唑蓝染色法（M1Tr）观察腺病毒mda-7／IL-24对乳腺癌细胞生长的影响；Hoechst染色和膜联蛋白（Annexin）V检测腺病毒mda-7／IL-24对乳腺癌细胞凋亡的影响；并在裸鼠上进行了动物实验研究。结果腺病毒mda-7／IL-24感染的乳腺癌细胞上清液中可以检测到mda-7／IL-24蛋白的表达；腺病毒mda-7／IL-24能明显地抑制乳腺癌细胞的生长，Hochest染色和AnnexinV提示腺病毒mda-7／IL-24促进乳腺癌细胞的凋亡。实验进一步证明了腺病毒mda-7／IL-24可以显著抑制裸鼠乳腺癌荷瘤的生长。结论腺病毒mda-7／IL-24能显著抑制乳腺癌细胞的生长并诱导乳腺癌细胞的凋亡。

Objective To investigate the effects of adenovirus melanoma differentiation associated gene-7 （mda-7）/IL-24 on the growth and apoptosis of human breast cancer cells. Methods Human breast cancer cells of the lines MCF-7 and MD-MBA-453 were cultured and transfected with purified adenovirns mda-7/IL-24. 48 h later, Western blotting was used to detect the protein expression of mda-7/IL-24 in the MCF-7 cells. The growth of Ad-mda-7/IL-24 in the MCF-7 cells was assayed by crystal violet staining and MTr method. The apoptotic effect of Ad-mda-7/IL-24 in the MCF-7 and MDA-MB-453 cells was detected by Hoeehst staining and annexin V method. Nude mice were inoculated with MCF-7 cells and randomly divided into 2 groups to be injecte4d with Ad-mda-7/IL-24 of PBS. The size of tumor was observed. Results Western blotting showed that mda-7/IL-24 was expressed in the transfected MCF-7 cells. Crystal violet staining and MTF method showed that Ad-mda-7/IL-24 dose and time-dependently inhibited the growth of the MCF-7 and MDA-MB-453 cells. Hochest staining and annexin V test indicated obvious apoptosis of the breast cancer cells. The size of tumor of the nude mice injected with Ad-mda-7/IL-24 was significantly smaller than that of the control mice. Conclusion Ad-mda-7/IL-24 effectively inhibits the growth of breast cancer cells and induces its apoptosis.