摘要：

目的:探讨蟾蜍灵联合顺铂对舌鳞状细胞癌(以下简称舌鳞癌) Tca8113细胞增殖,凋亡的影响及其机制.方法培养人舌鳞癌Tca8113细胞株,取对数期细胞分为对照组,蟾蜍灵组,顺铂组,联合组.对照组加入RP-MI 1640培养基;蟾蜍灵组分别加入10,20,40,80,160 nmol/L的蟾蜍灵;顺铂组分别加入2.5,5,10,20,40μg/mL的顺铂;联合组分别加入10,20,40,80,160 nmol/L的蟾蜍灵后按浓度对应加入2.5,5,10,20,40μg/mL的顺铂;MTT法检测各组细胞增殖抑制率,计算抑制细胞增殖50%的药物浓度( IC50)及联合指数( CI).根据蟾蜍灵与顺铂48 h时的IC50选择三个药物组相应浓度亚组,即蟾蜍灵40 nmol/L组(蟾蜍灵亚组),顺铂10μg/mL组(顺铂亚组),蟾蜍灵40 nmol/L,顺铂10μg/mL组(联合亚组),流式细胞术分析各亚组细胞凋亡情况,Western-Blot法检测各亚组Bax,Bcl-2,Caspase-3,细胞外调节蛋白激酶( Erk)及其磷酸化细胞外调节蛋白激酶状态( P-Erk)的表达.结果与对照组相比,蟾蜍灵组,顺铂组,联合组细胞增殖受到抑制,且联合组增殖抑制率高于蟾蜍灵组与顺铂组,并呈时间-剂量依赖关系(P均<0.05).联合组联合指数(CI)均小于1.联合亚组细胞凋亡率高于蟾蜍灵亚组与顺铂亚组,三组与对照组相比,P均<0.05.蟾蜍灵亚组,顺铂亚组,联合亚组Bax,Caspase-3蛋白表达高于对照组,其中联合亚组高于蟾蜍灵亚组和顺铂亚组;三个药物亚组P-Erk蛋白表达均低于对照组,其中联合亚组低于蟾蜍灵亚组和顺铂亚组;联合亚组Bcl-2表达低于对照组及蟾蜍灵亚组和顺铂亚组(P均<0.05).结论蟾蜍灵与顺铂单用或联合应用均可抑制Tca8113细胞增殖,促进其凋亡,联用效果更强;其作用机制可能与调节细胞凋亡相关蛋白表达,抑制Erk信号通路有关.%Objective To investigate the effects of bufalin and cisplatin on proliferation and apoptosis of tongue squa -mous cell line Tca8113.Methods Tongue squamous cell line Tca8113 was cultured and the logarithmic phase cells were divided into control, bufalin(10,20,40,80,160 nmol/L), cisplatin(2.5,5,10,20,40 μg/mL) and combination groups. MTT assay was used to examine the inhibition rate of cell growth when cell were treated at various concentrations of bufalin and cisplatin alone or in combination and calculate inhibiting concentration of 50%cell growth ( IC50 ) .The corresponding concentrations of three drug groups were selected based on IC 50 of Bufalin and cisplatin at 48 h.Apoptosis was evaluated by flow cytometry.The protion expression of Bax ,Bcl-2,Caspase-3,P-ERK and Erk were examined by Western blot method . Results Bufalin or cisplatin effectively inhibited the growth of Tca 8113 cells in a concentration-dependent manner at cer-tain range of concentrations .The inhibitory rate of combined treatment with bufalin and cisplatin was significantly higher than that of bufalin or cisplatin alone (P<0.05).The combination index(CI) values were all <1.When the Tca8113 cells were exposed to bufalin (40 nmol/L) combined with cisplatin (10 μg/mL) for 24 h, apoptosis rates were significantly in-creased.The difference was significant between drug treatment groups and control group (P<0.05).Both of bufalin and cisplatin could decrease the protein expression of p-Erk and increase the protein expression of Bax and Caspase-3.All these effects were enhanced when identical experiments were treatment with combination of the two drugs .Bufalin or cisplatin a-lone did not affect the protein expression of Bcl-2 obviously , but it was significantly decreased when combination used ( P<0.05).Conclusions Bufalin combined with cisplatin has synergistic effects in inhibiting the growth and inducing apopto -sis of Tca8113 cells and regulating the proteins expression of apoptosis-related and inhibiting Erk signaling pathway may be the related mechanism .

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