摘要
目的:研究人骨形态发生蛋白2和9(BMP2和BMP9)对人胃癌MNK-45细胞增殖、凋亡和迁移的影响,并初步探讨其作用机制。方法:用重组腺病毒AdBMP2和AdBMP9感染MNK-45细胞,采用MTT法、Hoechst 33258染色、流式细胞术(FCM)、划痕愈合实验和Transwell小室检测BMP2和BMP9对MNK-45细胞增殖、凋亡和迁移能力的影响,Western blotting检测总GSK-3β、磷酸化GSK-3β(p-GSK-3β)及β-catenin的表达水平。结果:(1)MTT显示AdBMP2和AdBMP9转染后,MNK-45细胞的增殖自第3 d开始被抑制并呈时间依赖性;(2)Hoechst 33258染色与FCM一致显示上调BMP2和BMP9表达可以促进MNK-45细胞的凋亡;(3)划痕愈合与Transwell迁移实验结果一致提示上调BMP2和BMP9表达均可抑制MNK-45细胞的迁移;(4)Westernblotting显示BMP2组和BMP9组的p-GSK-3β均较GFP组升高,但对β-catenin的表达量无明显影响(P>0.05);上述结果与对照组相比差异均有统计学意义(P<0.05或P<0.01)。结论:上调BMP2和BMP9表达对MNK-45细胞的增殖具有抑制作用,其机制与β-catenin无明显关系。
AIM ： To investigate the effects of human bone morphogenetic protein 2 （BMP2） and BMP9 on the proliferation, apoptosis and migration of human gastric carcinoma cell line MNK - 45. METHODS ： Immunocytochemical staining, MTT assay, wound -healing test, Transwells migration test, Hoechst 33258 staining and flow cytometry （FCM） were used to determine the infection of AdBMP2 and AdBMP9 on the proliferation, apoptosis and migration of MNK - 45 cells. The expression of GSK - 3β （ including p - GSK - 3β and total GSK - 3β） and β - catenin in MNK - 45 cells was also detected by Western blotting. RESULTS ： The proliferation of MNK - 45 cells was inhibited from the third day on and in a time - dependent manner after infected with AdBMP2 and AdBMP9. The results of Hoechst 33258 staining and FCM proved that apoptosis rates in BMP2 group and BMP9 group were higher than that in GFP group. Both wound - healing test and Transwell experiment indicated that up - regulating the expression of BMP2 and BMP9 inhibited the migration of MNK -45 cells. The phosphorylation levels of GSK -3β in BMP2 group and BMP9 group were higher than that in GFP group. However, no significant change of β - catenin among groups was observed. CONCLUSION： Up - regulation of BMP2 and BMP9 expression inhibits the proliferation of MNK -45 cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2012年第12期2160-2166,共7页
Chinese Journal of Pathophysiology
