张岩 乔录新 曾静 徐军 石英

    [摘要] 目的 探讨碳酸氢钠(NaHCO3)对肝癌生长的影响及其机制。 方法 5只7周龄裸鼠于无特定病原体(SPF)条件下的层流架中饲养，皮下注射肝癌细胞株HepG2细胞构建肝癌模型，并通过肿块内局部注射5%NaHCO3，体内研究肝癌肿块生长及组织坏死。Calcein/PI染色和流式细胞技术体外研究不同酸碱环境和NaHCO3作用下HepG2凋亡情况。蛋白印迹技术检测NaHCO3作用下HepG2细胞凋亡调控蛋白Bax/bcl2表达水平。 结果 5只裸鼠肝癌5%NaHCO3局部注射4周，3只肿块明显缩小，1只肿块长大，1只小鼠实验过程中死亡。5%NaHCO3 3/4、1/2、1/4、1/8、1/16和0(对照)培养基体积占比，Calcein/PI染色显示HepG2细胞凋亡率依次减低。流式细胞分析显示5%NaHCO3 1/2、1/4、1/8、1/16体积占比和对照组细胞凋亡率也依次减低。Calcein/PI染色显示培养基pH6.0、pH6.25、pH6.5、pH6.75、pH7.0、pH7.25、pH7.5、pH7.75、pH8.0的HepG2细胞凋亡率在pH7.0～7.25最低，随着酸碱度的增加凋亡率逐渐增加。流式细胞分析培養基pH6.25、pH6.5、pH6.75、pH7.0、pH7.25和pH7.5组细胞凋亡率变化显示类似结果。5%NaHCO3占1/4培养基体积比条件下HepG2细胞促凋亡的Bax mRNA和蛋白水平与对照相比较无明显变化，而抑凋亡的bcl2 mRNA和蛋白水平明显低于对照组。 结论 NaHCO3可通过改变肝癌组织内环境酸碱度，继而抑制凋亡功能蛋白blc2诱导肝癌细胞凋亡。

    [关键词] 碳酸氢钠；肝细胞癌；凋亡；治疗

    [中图分类号] G735.7 [文献标识码] A [文章编号] 1673-7210(2018)08(b)-0009-04

    [Abstract] Objective To explore the apoptotic mechanism of hepatocellular carcinoma (HCC) induced by sodium bicarbonate (SB). Methods HepG2 cells were implanted the subcutaneous of 5 athymic mice (7-week age) to form hepatocarcinoma mouse model and the growth and necrosis of hepatocellular carcinoma was observed under the condition of local mass injection of 5% SB. The apoptosis of hepatocellular carcinoma cell line HepG2 was observed by Calcein/PI staining and flow cytometry at SB exposure and different pH gradients. The expression of Bax/bcl2 were also detected in HepG2 cells under SB exposure by real-time PCR and western blotting. Results Animal in vivo studies revealed that 5% SB local injection for 4 weeks could inhibit tumor growth and cause tumor necrosis in 3 of 5 hepatocarcinoma mice. One hepatocarcinoma mice died and another mice had an enlarged tumor. 5% SB was diluted to the volume ratio 3/4 ......