核转位序列敲除的人类aFGF对NIH3T3细胞的促分裂活性及其机制  

Effect of aFGF with nuclear translocation sequence-knocked out on mitogenic activity to NIH3T3 cells and its mechanism

作　　者：吕文天[1] 龚平生[2] 王志成[1] 赵红光[1] 姜晓燕[1] 郭伟[1] 李艳博[1] 李校堃[3] 龚守良[1] 

机构地区：[1]吉林大学公共卫生学院卫生部放射生物学重点实验室,吉林长春130021 [2]吉林大学分子酶学工程教育部重点实验室,吉林长春130023 [3]温州医学院生物与天然药物研究院,浙江温州325035

出　　处：《吉林大学学报（医学版）》2006年第5期745-749,共5页Journal of Jilin University：Medicine Edition

基　　金：国家863专题项目资助课题(2002AA2Z3318)

摘　　要：目的：研究核转位序列敲除的人类酸性成纤维细胞生长因子（aFGF）对NIH3T3细胞促分裂活性的影响。方法：实验分4组，即野生型aFGF（waFGF）、重组型aFGF（raFGF）、waFGF加肝素（HS，waFGF＋HS）和raFGF＋HS。采用MTT法检测NIH3T3细胞增殖，通过流式细胞术检测NIH3T3细胞凋亡，采用RT—PCR法检测bcl-2、p53和c-los基因的表达。结果：当raFGF或waFGF浓度10～40μg·L^-1时，raFGF组细胞的增殖作用明显低于waFGF组（P〈0．001）；waFGF＋HS组和raFGF＋HS组均明显高于单纯waFGF或raFGF组（P〈0．001）。waFGF和raFGF均可导致NIH3T3细胞凋亡百分数下降，即存活的细胞增多，但raFGF作用弱于waFGF。c-los基因在waFGF和waFGF＋HS组的表达量均高于raFGF和raFGF＋HS组；p53基因表达在waFGF和raFGF两组表达增高；bcl-2基因在waFGF＋HS组表达最高，其次是waFGF和raFGF两组。结论：与waFGF比较，raFGF具有较弱的促分裂活性，但仍具有刺激DNA合成和细胞增殖的能力及抑制NIH3T3细胞凋亡的作用。HS具有增强aFGF活性的作用，可作为aFGF体外实验中模仿机体内环境的辅助因子。Objective To study the effect of aFGF with the nuclear translocation sequence-knocked out on the mitogenic activity to NIH3T3 cells. Methods There were four groups in the experiment; waFGF, raFGF, waFGF ＋ HS and raFGF＋HS. The cell proliferation was measured with MTT and the cell cycle progression and apoptosis were measured with FCM. The expressions of bcl-2, p53 and c-los genes were determined with RT-PCR. Results As the concentrations of raFGF or waFGF were at 10 - 40 μg · L^-1 , the effect of raFGF on the cell proliferation was significantly lower than that of waFGF （P〈0. 001）; when HS was added, the cell proliferations in waFGF＋ HS and raFGF＋ HS groups were significantly higher than those in waFGF and raFGF groups, respectively （P〈0. 001）. The apoptotic percentages of NIH3T3 cells treated with aFGF and raFGF all decreased, but the effect of raFGF was weaker than that of waFGF. The expressions of c-los gene in waFGF and waFGF＋ HS groups were higher than those in raFGF and raFGF＋ HS. The expressions of p53 gene in waFGF and raFGF groups increased. The expression of bcl-2 in waFGF ＋ HS group was highest, next were those in waFGF and raFGF groups. Conclusion The mitogenic activity of raFGF is weaker than that of waFGF. They all can stimulate DNA synthesis and cell proliferation, and inhibit the NIH3T3 cell apoptosis. Heparin can enhance the aFGF activity, and act as a asistant factor to imitate the the internal environment in aFGF in vitro.

关 键 词：成纤维细胞生长因子1 肝素 细胞增殖 细胞凋亡 基因表达 

分 类 号：Q7[生物学—分子生物学]