一种肠道微生物代谢产物混合物对肝癌的影响  

A metabolite cocktail of gut microbiota affects hepatocellular carcinoma

作　　者：董妍汐 王滨 张书琴 肖惠文 刘杏忠 董佳丽[1] 崔明[1] DONG Yanxi;WANG Bin;ZHANG Shuqin;XIAO Huiwen;LIU Xingzhong;DONG Jiali;CUI Ming(Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine,Institute of Radiation Medicine,Chinese Academy of Medical Sciences&Peking Union Medical College,Tianjin 300192,China;Department of Microbiology,College of Life Sciences,Nankai University,Tianjin 300071,China)

机构地区：[1]中国医学科学院北京协和医学院放射医学研究所,天津市放射医学与分子核医学重点实验室,天津300192 [2]南开大学生命科学学院微生物学系,天津300071

出　　处：《微生物学报》2024年第9期3409-3418,共10页Acta Microbiologica Sinica

基　　金：天津市杰出青年科学基金(20JCJQJC00100);国家自然科学基金(82373524,32100087,82202944)。

摘　　要：【目的】探究肠道微生物代谢产物的混合物——吲哚-3-丙酸(indole-3-propionic acid,IPA)、丁酸钠(sodium butyrate,SB)和戊酸(valeric acid,VA)对肝癌细胞增殖的影响。【方法】体外培养人肝癌HepG2细胞系,分别给予不同浓度的混合物(1×、2×、3×、4×和5×)处理细胞,用总胆固醇和甘油三酯检测试剂盒检测细胞总胆固醇(total cholesterol,TC)和甘油三酯(triglycerides,TG)含量,用细胞增殖-毒性检测试剂盒(Cell Counting Kit-8,CCK-8)及克隆形成检测细胞增殖。12只BALB/c无胸腺裸鼠随机分为对照(Ctrl)组和处理(Treat)组,皮下注射HepG2细胞。每3d测量肿瘤大小并计算肿瘤体积和抑瘤率。当肿瘤体积达到100 mm^(3)时,Ctrl组小鼠每天给予无菌水灌胃,Treat组每天给予混合物灌胃直至麻醉处死。治疗27d后,分别称量两组小鼠的体重,分离肿瘤并称量瘤重,计算瘤重/体重比。通过免疫组织化学(immunohistochemical,IHC)染色比较肿瘤Ki-67蛋白的含量,通过油红O染色比较肿瘤组织内的脂质积累。【结果】IPA、SB、VA的混合物降低了肝癌HepG2细胞内的TC和TG,对HepG2细胞的增殖有抑制作用,CCK-8和克隆形成结果都表明,混合物以剂量依赖的方式抑制HepG2细胞的增殖。在体内,混合物灌胃阻断了肝癌细胞的生长,表现为更小的肿瘤体积,更轻的肿瘤质量和更低的瘤重/体重比(Ctrl组:723mm^(3),0.47g,22.23%;Treat组:526 mm^(3),0.32 g,16.65%)。IHC和油红O染色进一步证明Ki-67的表达和脂质积聚在混合物处理的小鼠中减少。【结论】IPA、SB、VA的混合物可以抑制肝癌细胞增殖,抑制脂质合成。[Objective]To investigate the effects of a metabolite cocktail composed of indole-3-propionic acid(IPA),sodium butyrate(SB),and valeric acid(VA)of gut microbiota on the proliferation of hepatocellular carcinoma cells.[Methods]The human hepatocellular carcinoma HepG2 cells were cultured in vitro and treated with the cocktail at different concentrations(1×,2×,3×,4×,and 5×).The total cholesterol(TC)and triglyceride(TG)levels in the cells were determined by the total cholesterol and triglyceride assay kits.The Cell Counting Kit-8(CCK-8)and colony formation assays were employed to examine the cell proliferation.Twelve BALB/c athymic nude mice were randomized into a control(Ctrl)group and a treatment(Treat)group and then subjected to subcutaneous injections of HepG2 cells.The tumor size was measured every three days,and the tumor volume and tumor inhibition rate were calculated.When the tumor volume reached 100 mm^(3),the mice in the Ctrl group were administered with sterile water by gavage daily,while those in the Treat group received the cocktail via gavage until euthanized under anesthesia.After 27 days of treatment,the body weights of mice in both groups were measured,and tumors were excised and weighed,with the tumor weight/body weight ratio calculated.The content of Ki-67 protein in the tumors was determined by immunohistochemical(IHC)staining,and the lipid accumulation within tumor cells was assessed by Oil Red O staining.[Results]The cocktail of IPA,SB,and VA lowered the levels of TC and TG in hepatocellular carcinoma HepG2 cells and exerted an inhibitory effect on the proliferation of HepG2 cells.Both CCK-8 and colony formation assays indicated that the cocktail inhibited the proliferation of HepG2 cells in a dose-dependent manner.The oral administration of the cocktail inhibited the growth of hepatocellular carcinoma cells,as evidenced by smaller and lighter tumors and lower tumor weight/body weight ratios in the Treat group than in the Ctrl group(Ctrl:723 mm^(3),0.47 g,22.23%;Treat:526 mm^(3),0.32 g,16.

关 键 词：肠道微生物代谢产物 肝癌 细胞增殖 脂质合成 

分 类 号：R735.7[医药卫生—肿瘤]