单用Acivicin和联用顺铂诱导HepG_2细胞凋亡的实验研究

目的 :观察Acivicin单用及联用顺铂后对HepG2 细胞凋亡及相关基因Bcl -2、c -myc、p53表达的影响。方法 :Acivicin和顺铂分别或联用处理HepG2 细胞 ,采用MTT法测定细胞毒性 ;采用细胞凋亡原位检测法检测细胞凋亡率 ;采用免疫组化法检测凋亡相关基因Bcl -2、c -myc、p53 的表达。结果 :Acivicin为0 28、0 56、0 84、1 12、1 40mmol/L时 ,HepG2 细胞的凋亡率分别为(3±1 25) %、(3 7±2 0) %、(10±1 25) %、(17 4±4 5) %、(16 9±3 5) %。Acivicin(1 40mmol/L)在24、48、72h致HepG2 细胞的凋亡率分别为 (16 9±3 5) %、(27 9±4 3) %、(47 2±3 0) %,与对照组相比差异显著 (P<0 05) ;顺铂 (67μmol/L)致HepG2细胞 (24h)的凋亡率为 (73 4±1 5) %;Acivicin(1 40mmol/L)联用顺铂 (67μmol/L)致HepG2 细胞 (24h)的凋亡率为 (94 7±0 5) %,较两者单用显著增加 (P<0 01)。Bcl-2、c -myc、p53 检测结果显示 ,各组Bcl -2、c -myc、p53 的表达比对照组均有增加。结论 :Acivicin诱导HepG2 细胞凋亡呈剂量/时间依赖性 ;Acivicin与顺铂联用后可增强顺铂致HepG2 细胞凋亡的作用。

Experimental Study on the Apoptosis of HepG2 Cells Induced by Acivicin or/and Cisplatin

OBJECTIVE:To study the effects of Acivicin or/and cisplatin on the apoptpsis of HepG2 and expression of Bcl-2,c-myc and p53.METHODS:Acivicin and cisplatin were used to treat cultured HepG2 cell line separately or in combination.The cytotoxicity was measured by MTT assay,the apoptosis by TUNEL assay and relevant regulator genes by immunohistochemical SP method.RESULTS:Acivicin in concentrations of 0.28,0.56,0.84,1.12 and 1.40mmol/L,the apoptosis rates of HepG2 were (3±1.25)%,(3.7±2.0)%,(10±1.25)%,(17.4±4.5)%and (16.9±3.5)%respectively.Acivicin in concentration of 1.4mmol/L,the apoptosis rates at 24,48 and 72h were(16.9±3.5)%,(27.9±4.3)%and (47.2±3.0)%respectively,which was significantly different from control group.The apoptosis rate of HepG2 induced by cisplatin(67μmol/L)was (73.4±1.5)%at 24h.The apoptosis rate of HepG2 induced by Acivicin(1.4mmol/L)plus cisplatin(67μmol/L)was(94.7±0.5)%at 24h,which was markedly higher than those induced by Acivicin or cisplatin alone.The expressions of Bcl-2,c-myc and p53 in all groups were increased comparing with those in control group.CONCLUSION:This study indicates that the apoptosis of HepG2 induced by Acivicin was in a dose-dependent and time-dependent manner.When Acivicin and CDDP used simultaneously,the effect of CDDP on HepG2 apoptosis was enhanced greatly.