摘要
目的:探讨肝细胞生长因子(HGF)对肝癌细胞HepG2增殖和凋亡的影响.方法:人肝癌细胞株HepG2加入不同浓度HGF(0,1,10,50μg/L)培养2,4,6d,应用MTT法检测细胞增殖情况,流式细胞仪检测细胞周期及凋亡百分率的变化,免疫细胞化学法检测增殖细胞核抗原(PCNA)的表达.结果:HGF抑制HepG2细胞增殖呈剂量及时间依赖性,并诱导其凋亡[50μg/LHGF,培养时间为4,6d,其抑制率:(30.7±10.2)%,(49.8±11.1)%vs1μg/LHGF:(10.7±11.2)%,(4.2±9.4)%;P<0.05,P<0.01].随着HGF浓度增大S期细胞明显减少[试验组10,50μg/LHGF:(7.5±4.4)%,(7.8±1.6)%vs对照组0μg/LHGF:(16.6±2.8)%;P<0.05,P<0.01],同时可见G0/G1期细胞累积现象[试验组10,50μg/LHGF:(80.5±3.2)%,(73.1±3.9)%vs对照组0μg/LHGF:(59.6±6.2)%;P<0.05,P<0.01].试验组G1峰前出现明显的凋亡峰,凋亡率亦较对照组显著增加(P<0.05).且试验组细胞PCNA表达明显高于对照组.结论:HGF对肝癌细胞HepG2有抑制增殖和诱导其凋亡的作用,诱导细胞G0/G1期阻滞为可能机制之一.
AIM： To evaluate the effect of hepatocyte growth factor （HGF） on the proliferation and apoptosis in HepG2 cells. METHODS： HepG2 cells were cultured for 2, 4, 6 d in the presence of HGF at different concentrations of 0, 1, 10, 50μg/L. The effects of HGF on the proliferation of HepG2 cells was evaluated by MTT assay. The flow cytometry （FCM） was used to detect phase distribution of the cycles and apoptosis rate. Expression of proliferating cell nuclear antigen （PCNA） was detected with immunocytochemical technique. RESULTS： HGF inhibited the cell proliferation and induced the apoptosis in HepG2 in a time- and concentration-dependent manner. FCM also showed a sub-G1 cell apoptotic peak and a significant increace of apoptosis rate induced by HGF （P 〈 0. 05 ）. At the concentration of 50 Owe＇L, HGF treatments for 4 and 6 d depressed the proliferationof HepG2 cells by （30.7±10.2）% and （49.8±11.1）% , while at 1μg/L, HGF decreased the proliferation by （ 10.7±11.2） % and （4.2±9.4） %. Differences between the 2 groups were statistically significant （ P 〈 0. 05, P 〈 0.01 ）. DNA ploidy analysis showed that S phase cells were significantly decreased and quiescent G0/G1 phase cells were accumulated by raising HGF concentration. Percentages of S phase cells in 10 and 50μg/L groups were （7.5±4.4） %, （ 7.8±1.6 ） %, significantly lower than the control（ 16.6±2.8 ） % ; percentages of G0/G1 phase cells were （80.5±3.2）% and （73.1±3.9）% , significantly higher than the control （59.6±6.2 ） % （ P 〈 0. 05, P 〈 0.01 ）. Levels of PCNA were obviously down-regulated in HGF group compared with control group. CONCLUSION： HGF significantly inhibits the proliferation and induces the apoptosis in HepG2, which may be involved in G0/G1 phase arrest.
出处
《第四军医大学学报》
北大核心
2006年第8期717-719,共3页
Journal of the Fourth Military Medical University
