M-CSF上调cyclinD1/D3和CDK2/6的表达促进HeLa细胞增殖  

M-CSF Accelerates The Proliferation of HeLa Cells by Up-regulating The Expression of CyclinD1/D3 and CDK2/6

作　　者：涂剑[1] 吴海燕[1] 张蒙夏[2] 张晓红[2] 罗红梅[2] 龙治峰[2] 汪煜华[1] 雷小勇[1] 唐圣松[1,2] 

机构地区：[1]南华大学药物药理研究所,药物蛋白质组学湖南省高等学校重点实验室,衡阳421001 [2]南华大学组织胚胎学教研室,衡阳421001

出　　处：《生物化学与生物物理进展》2009年第7期910-915,共6页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30270684);湖南省自然科学基金资助项目(08JJ5004);湖南省教育厅科技计划项目(07A059;07C635);湖南省科技计划重点项目(06FJ3203)~~

摘　　要：非分泌型巨噬细胞集落刺激因子(M-CSF)的表达在肿瘤的发生发展过程中发挥重要作用,为探讨胞质M-CSF对细胞增殖的影响,采用基因重组技术构建胞内稳定表达M-CSF的HeLa细胞系,以空载体(pCMV/myc/cyto)转染HeLa细胞和未转染HeLa细胞作为对照,MTT法及反义寡核苷酸抑制实验分析M-CSF对细胞增殖的影响,并计算细胞倍增时间,RT-PCR观察胞内M-CSF对G1期细胞周期相关蛋白的影响.结果显示,与对照组比较,转染M-CSF的HeLa细胞倍增时间明显缩短、增殖能力显著增强,M-CSF的特异性反义寡核苷酸能抑制转染M-CSF的HeLa细胞的增殖,且抑制率随着反义寡核苷酸浓度的增高而增强,转染M-CSF的HeLa细胞的cyclinD1/D3和CDK2/6 mRNA表达显著升高(P<0.05).提示:M-CSF可上调cyclinD1/D3和CDK2/6的mRNA表达,促进HeLa细胞的增殖.Non-secreted macrophage colony-stimulating factor（M-CSF） plays an important role in genesis and progression of tumors. To explore the regulation of cytoplasmic M-CSF on the proliferation of HeLa cells, pCMV/myc/cyto-M-CSF vectors were transfected into HeLa cells. After comfirmed by RT-PCR, Western blot and immunocytochemistry, the effect of cytoplasmic M-CSF on the proliferation of HeLa cells were analyzed by MTT and antisense oligonucleotides. The doubling time was counted according to cell growth curves. The mRNA expression of cyclinE, cyclinD 1/2/3, CDK2/4/6 were assayed by RT-PCR. The results from RT-PCR, Western blot and immunocytochemistry showed that both M-CSF mRNA and protein were expressed at a higher level and localized to the cytoplasm in M-CSF-transfected HeLa cells, compared with either pCMV/myc/cyto-transfected HeLa cells or untransfected HeLa cells. M-CSF-transfected HeLa cells had shorter doubling time and more significantly augmented reproductive activity than either pCMV/myc/cyto-transfected HeLa cells or untransfected HeLa cells. M-CSF specific antisense oligonucleotides significantly inhibited the proliferation of the M-CSF-transfected cells, but had little effect on the other two groups. Furthermore, cytoplasmic M-CSF up-regulated the mRNA expression of cyclinDl, cyclinD3, CDK2 and CDK6（P 〈 0.05）. So it was concluded that cytoplasmic M-CSF accelerates the proliferation of HeLa cells by up-regulating the mRNA expression of cyclinD l/D3 and CDK2/6.

关 键 词：巨噬细胞集落刺激因子 HELA细胞 pCMV/myc/cyto-M-CSF载体 细胞增殖 细胞周期相关蛋白 

分 类 号：Q2[生物学—细胞生物学] R73[医药卫生—肿瘤]