bFGF对人肝癌细胞系Be1-7402的生长调控  

Effect of basic fibroblast growth factor on growth regulation in Bel-7402 cell line

作　　者：于卉影[1] 孙利于 孙黎光[1] 丁晓慧 

机构地区：[1]中国医科大学生物化学教研室,辽宁省沈阳市110001 [2]沈阳市第七人民医院内科,辽宁省沈阳市110003 [3]日本三重大学医学部解剖学

出　　处：《世界华人消化杂志》2003年第9期1333-1336,共4页World Chinese Journal of Digestology

基　　金：国家自然科学基金;No.39870384

摘　　要：目的:探讨bFGF是否通过PI3K/PKB途径调节p21^(WAFl)的表达。方法:^(32)P掺入法检测 PKB酶活性,RT-PCR、Western blot检测不同处理组Bel-7402细胞的p21^(WAFl)表达,流式细胞术分析细胞周期。结果:25 μg/L bFGF刺激细胞10min,就可使胞液和膜性PKB酶活性达高峰.p21^(WAFl)mRNA表达水平在1h达高峰,比对照升高了5.5倍.p21^(WAFl)蛋白表达在2 h达高峰,比对照升高了2.2倍.wortmannin预处理后,PKB活性在各时间点均明显降低(P<0.05),p21^(WAFl)mRNA表达及p21^(WAFl)蛋白表达无明显变化.流式细胞术分析显示,bFGF处理组与对照组相比 GI期细胞减少(0.65 ± 0.01→0.49 ± 0.02,P<0.01),S期细胞增多(0.14 ± 0.01→0.28 ± 0.01,P<0.01),wortmannin能抑制此促增生作用(GI:0.58±0.01;S:0.22 ±0.01,P<0.01)。结论:PI3K/PKB途径可介导bFGF对Bel-7402细胞的促增生作用,但是bFGF对p21^(WAFl)表达的调节作用不依赖PI3K/PKB途径。AIM: To investigate whether bFGF regulates p21^(WAF1) ex- pression of Bel-7402 cell line via PI3K/PKB pathway. METHODS: ^(32)P incompration assay. The expression of p21^(WAF1)- mRNA was assessed by RT-PCR. The expression of p21^(WAF1) protein was datected by Western blot. Cell Cycle analysis was performed on a FACScan. RESULTS: Both membrane and cytosol activity of PKB in Bel-7402 cell which were treated with bFGF (25 μg/L) reached the peak at 10 min. p21^(WAF1) mRNA level was upregulated and peaked at 1 h (5.5 fold induction). Correspondingly, p21^(WAF1) expression was increased and peaked at 2 h (2.2 folds of reduction). Wortmannin efficiently inhibited the activity of PKB (P<0.05), but not the level of p21^(WAF1) mRNA and the expression of p21^(WAF1) protein.FCM analysis showed bFGF induced S-phase entry (0.14±0.01→ 0.28±0.01, P<0.01), which was inhibited by wortmannin effectively (0.28±0.01→0.22±0.01, P<0.01). CONCLUSION: bFGF stimulates the proliferation of bel-7402 cell line via PI3K/PKB pathway, and modulates p21^(WAF1)ex- pression through separating signaling pathways.

关 键 词：BFGF 肝癌 Bel-7402细胞系 PKB酶 细胞周期 RT-PCR 流式细胞术 

分 类 号：R735.7[医药卫生—肿瘤]