生物钟基因Period2调节肝癌细胞warburg效应及转移侵袭中的作用机制

Effects of circadian clock gene Period2 on Warburg effect and metastasis in hepatocellular carcinoma cells

目的:研究钟基因Period2在肝癌细胞warburg效应及转移侵袭中的作用。方法:采用实时荧光定量PCR(RT-QPCR)检测组织以及肝癌细胞的Period2表达水平。上调肝癌细胞MHCC97L中Period2的水平,沉默HepG2中Period2的表达。检测葡萄糖摄取水平、乳酸水平、细胞氧耗水平、细胞活力、迁移能力、侵袭能力。Western-Blot检测细胞中代谢关键酶的表达。结果:肿瘤组织中Period2基因水平显著高于癌旁组织(P<0.05)。4株肿瘤细胞中MHCC97L中Period2表达水平较低,而HepG2水平较高。Period2沉默后,HepG2细胞葡萄糖相对摄取水平相比control下调、乳酸相对产生水平相比control下调,细胞耗氧量相比control增高,细胞活力、迁移及侵袭能力下降。糖代谢关键酶表达水平下调。而Period2过表达后,MHCC97L细胞葡萄糖摄取水平相比control增高、乳酸产生水平相比control上调,细胞耗氧量相比control降低,细胞活力、迁移及侵袭能力上调。糖代谢关键酶表达水平上调。结论:钟基因Period2在肝癌中高表达,其作用与促进肝癌细胞Warburg效应,促进转移侵袭有关。

AIM: To study the effects of circadian clock gene Period2 on Warburg effect and metastasis in hepatocellular carcinoma cells. METHODS: Real-time fluorescence quantitative PCR (RT-QPCR) was used to detect the expression of Period2 in tissues and 4 hepatocellular carcinoma cells,up-regulated the expression of Period2 in hepatocellular carcinoma cell line MHCC97L.silenced the expression of Period2 in hepatocellular carcinoma cell line HepG2.The glucose uptake level,lactic acid level,cell oxygen consumption level,cell viability,migration,and cell invasion were detected.Western-Blot was used to detect the expression of the key metabolic enzymes. RESULTS:The expression level of Period2 in MHCC97L was low,while HepG2 level was high.After Period2 silencing, the relative glucose uptake and lactate production of HepG2 cells were down-regulated compared with control group, and the oxygen consumption of HepG2 cells was increased compared with control,and the cell viability, migration and invasion ability were decreased.The expression level of key enzymes of glycometabolism was down-regulated.After overexpression of Period2,the glucose uptake level of MHCC97L cells was increased,the lactate production level was increased,the oxygen consumption was decreased compared with control, and the cell viability, migration and invasiveness were increased.The expression level of key enzymes of glucose metabolism was up-regulated. CONCLUSION: The clock gene Period2 is highly expressed in HCC, and its role is related to the promotion of Warburg effect and metastasis and invasion of HCC cells.