CT23通过调节H6PD表达影响磷酸戊糖途径调控肝细胞癌细胞凋亡
CT23 influences pentose phosphate pathway to regulate apoptosis of HCC cells by regulating H6PD expression
杨哲 1廖晓璐 2张若皓 3卢玉飞 3党德静 4宁敬予 2陈俊霏 2肖贝贝 2黄秒 5胡启平6
作者信息
- 1. 广西医科大学基础医学院细胞生物学与遗传学教研室,南宁 530021;广西高校生物分子医学研究重点实验室,南宁 530021
- 2. 广西医科大学第一临床医学院,南宁 530021
- 3. 广西医科大学基础医学院细胞生物学与遗传学教研室,南宁 530021
- 4. 广西医科大学第二临床医学院,南宁 530021
- 5. 广西医科大学附属肿瘤医院影像诊断中心,南宁 530021
- 6. 广西医科大学基础医学院细胞生物学与遗传学教研室,南宁 530021;广西医科大学长寿与老年相关疾病教育部重点实验室,南宁 530021
- 折叠
摘要
目的:探讨癌-睾丸抗原23(CT23)参与磷酸戊糖途径(PPP)调控,促进肝细胞癌(HCC)细胞凋亡的分子机制.方法:通过全转录组测序、葡萄糖消耗检测、乳酸生成分析、还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)生成检测、活性氧(ROS)生成分析和线粒体示踪等方法探讨CT23与PPP的关系;蛋白质印记法(western blotting)及实时荧光定量PCR(RT-qPCR)检测敲低CT23的HCC细胞中已糖-6-磷酸脱氢酶(H6PD)表达量变化,TUNEL法分析细胞凋亡.结果:CT23与PPP有关;与control组相比,shCT23组HCC细胞葡萄糖消耗减少,乳酸生成水平降低,NADPH生成水平降低,ROS水平升高,细胞凋亡增加,H6PD mRNA水平降低,H6PD蛋白水平降低(均P＜0.05);电镜下细胞形态发生变化并伴随线粒体损伤;与shCT23组相比,shCT23+ H6PDOE组HCC细胞H6PD蛋白水平升高,葡萄糖消耗增多,乳酸生成水平升高,NADPH生成水平升高,细胞凋亡减少(均P＜0.05).结论:CT23通过H6PD增强PPP促进HCC细胞凋亡.
Abstract
Objective:To explore the molecular mechanism by which cancer-testis antigen 23(CT23)partici-pates in the regulation of the pentose phosphate pathway(PPP)and promotes apoptosis in hepatocellular carcino-ma(HCC)cells.Methods:The relationship between CT23 and PPP was investigated by whole transcriptomic se-quencing,glucose consumption detection,lactic acid production analysis,reduced nicotinamide adenine dinucleo-tide phosphate(NADPH)production detection,reactive oxygen species(ROS)production analysis and mitochon-drial tracer analysis.Western blotting and reverse transcription-quantitative PCR(RT-qPCR)were used to detect the expression of hexose-6-phosphate dehydrogenase(H6PD)in HCC cells with CT23 knockdown,and the apop-tosis was analyzed by the TUNEL assay.Results:CT23 was related to PPP.Compared with the control group,HCC cells in the shCT23 group had decreased glucose consumption,decreased lactate production level,de-creased NADPH production level,increased ROS level,increased apoptosis,decreased H6PD mRNA level and H6PD protein level(all P＜0.05).The morphological changes of the cells were accompanied by mitochondrial damage under electron microscope.Compared with the shCT23 group,HCC cells in shCT23+H6PDOE group had increased H6PD protein level,increased glucose consumption,increased lactate production level,increased NADPH production,and decreased apoptosis(all P＜0.05).Conclusion:CT23 promotes HCC cell apoptosis through enhancing PPP via H6PD.
关键词
肝细胞癌/癌—睾丸抗原23/磷酸戊糖途径/己糖-6-磷酸脱氢酶Key words
hepatocellular carcinoma/cancer-testis antigen 23/pentose phosphate pathway/hexose-6-phosphate dehydrogenase引用本文复制引用
基金项目
国家自然科学基金(81760424)
中央引导地方科技发展专项(2022ZYZX1193)
广西自然科学基金(2024GXNSFAA010034)
广西医科大学大学生创业训练计划立项项目(2023)(202310598008)
出版年
2024
国家科技期刊平台
NSTL
万方数据