双特异性磷酸酶5对人胎盘滋养层细胞系BeWo增殖和凋亡的影响  

Effect of dual specificity phosphatase 5 on proliferation and apoptosis in human placental trophoblast cell line BeWo

作　　者：戴小燕[1] 李新哲[2] 黄团明 王婉[1] 俞丽丽[1] 郑英如[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所妇产科,重庆400042 [2]第三军医大学基础医学部生物化学与分子生物学教研室,重庆400038

出　　处：《第三军医大学学报》2014年第7期659-663,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(81070505)~~

摘　　要：目的探讨双特异性磷酸酶5(dual specificity phosphatase 5,DUSP5)基因过表达对人胎盘滋养层细胞系BeWo增殖和凋亡的影响及其可能机制。方法以携带人DUSP5基因的重组慢病毒感染BeWo细胞,用流式细胞仪分选建立过表达DUSP5的BeWo细胞模型,以携带空载体的慢病毒感染的细胞以及未经任何处理的细胞作对照。经荧光实时定量PCR和Western blot检测DUSP5基因mRNA和蛋白的表达,经CCK-8实验和流式细胞术检测BeWo细胞增殖和凋亡的变化,经Western blot检测磷酸化ERK(p-ERK)、p-p38及p-JNK的变化。结果①经荧光实时定量PCR和Western blot证实,成功构建了DUSP5过表达的人胎盘滋养层细胞BeWo细胞模型;②CCK-8实验结果显示DUSP5过表达组在第3、4、5天的光密度值D(490)与其他两对照组相比明显增高(P<0.01);流式细胞仪结果显示顺铂作用48 h后,DUSP5过表达组与空病毒载体组相比,正常细胞的百分比显著增加而早期凋亡细胞百分比则显著减少(P<0.01);Western blot结果显示DUSP5过表达组中p-ERK1/2水平显著下降,而p-p38和p-JNK未见明显变化。结论 DUSP5可促进人胎盘滋养细胞系BeWo增殖、抑制细胞凋亡,以上作用可能与DUSP5抑制ERK信号通路有关。Objective To determine the effect of the over-expression of dual specificity phosphatase 5 （DUSP5） on the proliferation, apoptosis in human placental trophoblast cell line BeWo and investigate the possible signaling pathway. Methods BeWo cells was infected by recombinant lentiviral vector carrying human DUSP5 gene. The cells infected by empty lentiviral vector and untreated cells served as control. Flow cytometry was used to select the cells over-expressing DUSP5. Real-time PCR and Western blotting were used to detect the mRNA and protein levels of of DUSP5. CCK-8 assay and flow cytometry were employed to detect cell proliferation and apoptosis. The expression of p-ERK1/2, p-p38 and p-JNK1/2/3 were detected by Western blotting. Results Real-time PCR and Western blotting indicated that the BeWo cells over-expressing DUSP5 was successfully established. CCK-8 assay confirmed that the BeWo cells over-expressing DUSP5 had signifi cantly higher rates of proliferation than the other 2 cells in days 3, 4 and 5 （P 〈0.01 ）. The number of normal cells was significantly increased, while that of early apoptotic cells was markedly lesser in DUSP5 over-expres sion cells than the vector group in 48 h after cisplatin treatment （P 〈 O. O1 ）. The protein level of p-ERK1/2 was lower in BeWo cells over-expressing DUSP5 than in cells, while that of p-p38 and p-JNK was not changed proliferation, and inhibits cell apoptosis in human inhibiting ERK1/2 signaling pathway. the cells infected by empty lentiviral vector and untreated cells, while that of p-p38 and p-JNK was not changed. Conclusion Over-expressing of DUSP5 induces cell proliferation, and inhibits cell apoptosis in human placental trophoblast BeWo cells, which may be relevant with inhibiting ERK1/2 signaling pathway.

关 键 词：双特异性磷酸酶5 BEWO细胞 细胞增殖 细胞凋亡 ERK1 2信号通路 

分 类 号：R321.4[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]