摘要
目的研究p21蛋白激活激酶4(p21protein activated kinase 4,PAK4)对人乳腺肿瘤细胞生长的影响。方法人乳腺肿瘤细胞MCF-7转染不同剂量PAK4真核表达载体或慢病毒敲除PAK4表达,收集细胞提取蛋白进行蛋白质印迹法检测PAK4和Cyclin D1表达。流式细胞仪检测PAK4过表达和敲除后对细胞生长周期影响。结果过表达PAK4明显上调人乳腺肿瘤细胞MCF-7中的Cyclin D1蛋白表达,引起G1期细胞减少(从80%下降到55%),导致G1/S期转化细胞增多。慢病毒敲除人乳腺肿瘤细胞MCF-7中的PAK4表达降低了的Cyclin D1蛋白表达,使G1期的细胞增加12%,而S期的细胞下调7%。结论 PAK4调节Cyclin D1表达,促进人乳腺肿瘤细胞MCF-7G1/S期转化,揭示PAK4可能是重要的人乳腺肿瘤治疗靶点。
OBJECIIVE To examine the influence of p21-activated kinases（PAK4） on the proliferation of human breast cancer cells MCF-7.METHODS Human breast cancer cells MCF-7 were treated with increasing concentrations of PAK4 constructs,and whole-cell lysates were extracted.Lysates containing equal amounts of protein were immunoblotted with PAK4 and Cyclin D1 antibody.Cell cycle was analyzed by flow cytometry assay.Inhibiting PAK4,by PAK4-RNAi-lentivirus,cell cycle was analyzed by flow cytometry assay.RESULTS The increased expression of PAK4 obviously upregulated the levels of Cyclin D1 protein and shortened G1 （down from 80 ％ to 55 ％） and accelerated the G1/S-phase transition.When human breast cancer cells MCF-7 were transfected with a shRNA-lentivirus against PAK4,the levels of Cyclin D1 protein were decreased and there was 12％ increase in G1 phase and 7％ reduction in S phase.CONCLUSIONS We showed a novel mechanism of modulation of cell proliferation by PAK4 through up-regulation of Cyclin D1 in human breast cancer cells MCF-7.PAK4 promotes G1/S transition in human breast cancer cells MCF-7,which is critical for the design of targeted therapies against human breast cancer.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2015年第3期165-168,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(31171360)
