肿瘤生酮代谢疗法敏感性依赖于酮体代谢酶水平  

The sensitivity of the ketogenic metabolic therapy in cancer depends on the expression of ketolytic key enzymes

作　　者：张杰 丛明华 高云[3] 俞伟男 缪明永[3] 石汉平 

机构地区：[1]徐州医科大学附属淮安医院淮安市第二人民医院内分泌科,江苏淮安223002 [2]国家癌症中心/中国医学科学院北京协和医学院肿瘤医院,北京100021 [3]第二军医大学基础医学部生物化学与分子生物学教研室,上海200433 [4]首都医科大学附属北京世纪坛医院胃肠外科/临床营养科,北京100038

出　　处：《肿瘤代谢与营养电子杂志》2017年第4期421-429,共9页Electronic Journal of Metabolism and Nutrition of Cancer

基　　金：国家自然科学基金(81570569);国家重点研发计划项目(2017YFC1309200);山东省抗体制药协同创新中心(聊城大学)开放课题项目(CIC-AD1804)

摘　　要：目的基础实验及临床研究均证实KD具有一定的抗肿瘤作用,但是疗效并不一致,尚没有特异性的分子靶标或明确的适应证来衡量其抗肿瘤作用。本研究拟检测多种肿瘤细胞系酮体代谢酶的表达情况,以验证不同酮体代谢酶表达水平的肿瘤细胞对KD治疗的敏感性。方法通过qRT-PCR检测33株肿瘤细胞系BDH1和OXCT1编码基因的表达情况。根据筛选结果选择BDH1和OXCT1高表达的HeLa细胞以及低表达PANC-1细胞进行KD敏感性的体内外验证。使用手持式细胞计数器检测含不同浓度βHB的低糖培养基中HeLa和PANC-1细胞增殖情况。然后,敲低HeLa细胞中BDH1和OXCT1的表达水平,构建HeLa和PANC-1裸鼠皮下移植瘤,体内外重新验证KD抗肿瘤治疗的敏感性。结果与对照组相比,低糖组HeLa细胞和PANC-1细胞增殖明显受抑制,但是低糖培养基加入βHB后,HeLa细胞增殖明显好转,但是PANC-1细胞增殖无显著好转。动物实验中,KD显著抑制PANC-1移植瘤的生长,但是对HeLa移植瘤的生长无抑制作用,而同时敲低BDH1和OXCT1的HeLa细胞对KD治疗敏感。结论酮体代谢关键酶BDH1和OXCT1低表达,并且无明显诱导的肿瘤对生酮代谢疗法敏感。Objective Although ketogenic diet （KD） has been studied its certain anti-tumor effect by basic research and clinicaltrials, the specific molecular targets or indications which verify its anti-tumor activity are lack. In this study, we examined theexpression of different ketolytic key enzymes to identify the sensitivity of KD in tumor cells. Methods Expression of genes encoding3-hydroxybutyrate dehydrogenase 1 （BDH1） and succinyl-CoA： 3-oxoacid CoA transferase 1 （OXCT1） have been determined in 33human cancer cell lines by quantitative RT-PCR. HeLa cell and pan-1 cell were selected with high expressing and low expressingtwo enzymes respectively. Proliferation of HeLa and PANC-1 cells was determined by using a hand-held automatic cell counter aftercultured in low glucose medium with or without DL-β-Hydroxybutyric acid sodium salt （βHB）. BDH1 and OXCT1 were knockeddown in HeLa cells by lentivirus-mediated RNA interference and established HeLa and PANC-1 cell xenograft tumors models inBALB/C nude mice to identify the sensitivity of KD in anti-tumor therapy. Results Compared to control group, proliferation ofHeLa and PANC-1 cells in low glucose （LG） was significantly inhibited. Howevere, proliferation of HeLa cells was significantlyincreased, while added βHB into LG group, but there was no significant effect on proliferation of PANC-1 cells. Animal experimentsdemonstrated that KD inhibited growth of PANC-1 cell xenograft tumors dramatically, but no effect on the growth of HeLa xenografttumor. Down-regulation of both BDH1 and OXCT1 in HeLa cells rendered their sensitivity to KD in vitro and in vivo. ConclusionsTumors with low expression of ketolytic enzymes which are not obviously induced gene expression are sensitive to ketogenicmetabolic therapy.

关 键 词：生酮饮食 肿瘤 酮体 代谢 

分 类 号：R730.5[医药卫生—肿瘤]