单-ADP-核糖基转移酶3过表达对人乳腺癌细胞MDA-MB-231增殖、凋亡及侵袭的影响

Effects of overexpression of mono-ADP-ribosyltransferase 3 on proliferation,apoptosis and invasion of human breast cancer MDA-MB-231 cells

目的探讨单-ADP-核糖基转移酶3(mono-ADP-ribosyltransferase 3,ART3)过表达对人乳腺癌MDA-MB-231细胞增殖、凋亡及侵袭的影响。方法在Lipofectamine 2000的介导下,将ART3基因重组真核表达质粒pCMV-ART3-myc-tag转染人乳腺癌细胞MDA-MB-231,用G418进行稳定转染细胞株的筛选,同时设未转染组对照。Western blot法检测稳定转染细胞中ART3-myc-tag蛋白的表达,MTT法、流式细胞术和Transwell试验分别检测ART3对MDA-MB-231细胞增殖活力、凋亡及侵袭能力的影响。结果 ART3可在MDA-MB-231细胞中稳定过表达;ART3转染组细胞的增殖活力和侵袭能力均明显高于未转染组(P<0.05);ART3转染组细胞的凋亡率明显低于未转染组(P<0.05)。结论过表达ART3能促进乳腺癌细胞增殖、侵袭,并抑制其凋亡,本实验为进一步研究ART3基因在乳腺癌发生发展中的作用及其机制奠定了基础。

Objective To investigate the effects of overexpression of mono-ADP-ribosyltransferase 3 （ART3） on prolifera- tion, apoptosis and invasion of human breast cancer MDA-MB-231 cells. Methods MDA-MB-231 cells were transfected with recombinant plasmid pCMV-ART3-myc-tag in medication of Lipofectamine 2000, using those untransfected as control. The stable transfectants were screened with G418 and determined for expression of ART3 by Western blot, for proliferative activity by MTY assay, for invasion ability by Transwell assay and for apoptosis by flow cytometry. Results ART3 was stably overexpressed in MDA-MB-231 cells. Both the proliferation activity and invasion ability of MDA-MB-231 cells transfected with recombinant plasmid pCMV-ART3-myc-tag were significantly higher （P 〈 0. 05 ）, while the apoptosis rate was significantly lower, than those untransfected （P 〈 0. 05 ）. Conclusion The overexpression of ART3 promot- ed the proliferation and invasion while inhibited the apoptosis of breast cancer cells, which ladi a foundation of further study on role of ART3 in the tumorigenesis and development of breast cancer.