摘要
目的 探讨人趋化素样因子 2 (CKLF2 )对小鼠成纤维细胞BALB c 3T3增殖和凋亡的影响。方法 构建pcDI CKLF2真核表达载体 ;建立稳定表达人CKLF2的BALB c 3T3(3T3 CKLF2 )细胞系 ;以细胞计数和MTT法分析过表达人CKLF2后对 3T3细胞增殖的影响 ;以流式细胞术检测annexin Ⅴ阳性细胞为凋亡指标 ,分析 3T3 CKLF2细胞和 3T3 pcDI细胞对撤除血清诱导凋亡的反应情况。结果 成功地建立了稳定表达人CKLF2的BALB c 3T3细胞系 ,过量表达人CKLF2的 3T3细胞系增殖速度明显加快 ,同等接种量的细胞培养 40h以后 ,3T3 CKLF2细胞的数量 (1.0 7× 10 6 )比 3T3 pcDI细胞(2 .5 4× 10 5)及野生型 3T3细胞 (3 .0× 10 5)高 2～ 3倍 ;流式细胞术分析表明 ,撤除血清后 3T3 CKLF2的annexin Ⅴ阳性细胞数 (13.34% 48h、15 .44 % 96h)明显低于 3T3 pcDI细胞 (19.36 % 48h、39.2 % 96h)。结论 人CKLF2能够促进BALB c 3T3细胞的增殖和抵抗撤除血清诱导的细胞凋亡 ,这些研究为进一步探讨CKLF家族的生物学功能提供了实验依据。
Objective To study the effect of human chemokine-like factor 2(CKLF2)on the proliferation and cell survival of BALB/c 3T3 cells. Methods We constructed a eukaryotic expression vector of pcDI/CKLF2 and established two cell lines, including CKLF2-overexpressed cell line (3T3/CKLF2) and a control cell line(3T3/pcDI). Cell proliferation was analyzed by MTT and cell counting assay; cell apoptosis after serum-withdrawal was analyzed by annexin-Ⅴ staining. Results We have successfully established two cell lines, 3T3/CKLF2 and 3T3/pcDI, and identified expression of CKLF2 by RT-PCR; 3T3/CKLF2 cells grow faster than the control cells, after cell culture for 40h, the number of 3T3/CKLF2 is 2-3 times more than that of 3T3/pcDI and wild type cells. Flow cytometry analysis showed that annexin-Ⅴ positive cells in 3T3/CKLF2 were less than those in 3T3/pcDI cells after serum withdrawal. Conclusion CKLF2 could promote proliferation and exert an anti-apoptotic effect on BALB/c 3T3 cells. These results provide experimental evidences for further study of biological activity of CKLF family. [
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2003年第1期5-8,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金 (No .30 0 0 0 15 3)
国家 86 3基金( 2 0 0 1AA2 15 0 6 1)资助项目
