二甲双胍联合MCT1抑制剂α-氰基-4-羟基苯乙烯治疗小鼠Lewis肺癌的研究

Study on the Anti-Lewis Lung Carcinoma Effect of Metformin Combined with MCT1 Inhibitor CHC

目的探讨二甲双胍联合单羧酸转运蛋白1(monocarboxylate transporter 1,MCT1)抑制剂α氰基-4-羟基苯乙烯(CHC)干扰乳酸代谢是否有抗肿瘤作用。方法体外检测对照组、二甲双胍组(1mmol/L、5mmol/L)、CHC组(5mmol/L)及联合用药组(二甲双胍5mmol/L和CHC 5mmol/L)24、48hLL/2细胞的增殖、乳酸代谢的改变及24h细胞凋亡变化。C57BL/6小鼠于右背侧皮下接种LL/2肿瘤细胞(5×105/只),建立小鼠Lewis肺癌模型,将28只荷瘤小鼠随机平均分为4组:对照组(0.1mL生理盐水灌胃,同时给予0.1mL生理盐水腹腔注射)、二甲双胍治疗组(二甲双胍灌胃,200mg/kg,0.1mL/只,同时给予0.1mL生理盐水腹腔注射)、CHC治疗组(CHC腹腔注射,100mg/kg,0.1mL/只,同时给予0.1mL生理盐水灌胃)和联合治疗组(同时给予上述剂量二甲双胍灌胃和CHC腹腔注射)。观察各组小鼠肿瘤生长、测量体积大小并进行肿瘤组织原位凋亡检测。结果在体外,联合使用二甲双胍和CHC可以显著增加肿瘤细胞的乳酸生成,同时抑制肿瘤细胞的增殖,与对照组、二甲双胍组和CHC组相比,差异均有统计学意义(P<0.05)。体内动物模型证实联合使用二甲双胍和CHC可以早期减缓肿瘤的生长,与对照组、二甲双胍组和CHC组比较,差异均有统计学意义(P<0.05);同时促进肿瘤细胞的凋亡。结论二甲双胍联合CHC可以改变肿瘤细胞乳酸代谢,诱导细胞凋亡,具有一定的抗肿瘤作用。

Objecitve To investigate the antitumor effect of the combination of metformin with α-cyano-4- hydroxycinnamic acid （CHC, a MCT1 inhibitor） in the treatment of Lewis lung cancer. Methods In vitro, the utilization of lactate acid was measured by lactate assay in cultured medium and the inhibition of LL/2 cell proliferation of four groups Ccontrol group, metformin group （1 mmol/L and 5 mmol/L）, CHC group （5 mmol/L） and the combination group （metformin 5 mmol/L and CHC 5 mmol/L）3 was detected in 24 h, 48 h, and their apoptosis in 24 h was also detected. In vivo, twenty eight C57BL/6 mice bearing LL/2 （5×10^5） subcutaneous Lewis lung cancer on the right flank was established and then randomly assigned into four groups： control, metformin （200 mg/kg body mass in 0.1 mL i.g. with NS 0.1 mL i. p. ）, CHC （100 mg/kg body mass in 0.1 mL i.p. with NS 0.1 mL i. g. ） and the combination （metformin 200 mg/kg body mass in 0.1 mL i.g. with CHC 100 mg/kg body mass in 0. 1 mL i. p. ）. Tumor volume was measured. The pathologic observation and apoptotic analysis of tumors was assessed by TUNEL assay. Results Compared to the contorl, metformin or CHC alone, combination of two drugs leaded to a significant lactate acid production in cultured medium and the inhibition of LL/ 2 cell viability （P〈0. 05）. In vivo, the systemic administration of two drugs leaded to obvious retarded tumor growth compared with used alone in early stage （P〈0.05）. The TUNEL assay showed the significantly increased number of apoptosis ceils in tumor tissues from the combination group. Conclusion Combination of metformin and CHC transformed the lactic acid metabolism in LL/2 cells and induced cell apoptosis and showed the antitumor effect.