吉林大学学报（医学版）2024，Vol.50Issue(3)：682-688,7.DOI:10.13481/j.1671-587X.20240312
灵芝酸A对非小细胞肺癌PC9细胞糖酵解及其关键限速酶的影响
Effect of ganoderic acid A on glycolysis and its key rate-limiting enzymes of non-small cell lung cancer PC9 cells
摘要
Abstract
Objective:To discuss the effects of different doses of ganoderic acid A(GAA)on the biological activities,glycolysis,and the expression of rate-limiting enzymes of non-small cell lung cancer(NSCLC)PC9 cells,and to clarify the mechanism.Methods:The NSCLC PC9 cells were cultured in vitro and divided into blank control group,low dose(25 µmol·L-1)of GAA group,medium dose(50 µmol·L-1)of GAA group,and high dose(100 µmol·L-1)of GAA group.The methylthiazolydiphenyltetrazolium(MTT)assay was used to detect the survival rates of the PC9 cells in various groups;Transwell chamber assay was used to detect the number of migration cells of the PC9 cells in various groups;the glucose uptakes of the PC9 cells in various groups were detected by glucose assay kit;the levels of ATP in the PC9 cells in various groups were detected by ATP assay kit;the levels of lactic acid in the PC9 cells in various groups were detected by lactate assay kit;the expression levels of hexokinase 2(HK2)and pyruvate kinase M2(PKM2)mRNA in the PC9 cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method;the expression levels of HK2 and PKM2 proteins in the PC9 cells in various groups were detected by Western blotting method.Results:The MTT assay results showed that,at 24,48,and 72 h of culture,compared with blank control group,the survival rates of the cells in medium and high doses of GAA groups were significantly decreased(P<0.05).At 72 h of culture,compared with low dose of GAA group,the survival rate of the cells in high dose of GAA group was significantly decreased(P<0.05).The Transwell chamber assay results showed that compared with blank control group,the numbers of migration cells in medium and high doses of GAA groups were significantly decreased(P<0.05);compared with low doses of GAA group,the number of migration cells in high dose of GAA group was significantly decreased(P<0.05).Compared with blank control group,the glucose uptakes and levels of lactic acid in the cells in medium and high dose of GAA groups were significantly decreased(P<0.05),and the level of ATP in the cells in high dose of GAA group was significantly decreased(P<0.05).The RT-qPCR results showed that compared with blank control group,the expression levels of HK2 and PKM2 mRNA in the cells in medium and high doses of GAA groups were significantly decreased(P<0.05).The Western blotting results showed that compared with blank control group,the expression levels of HK2 and PKM2 proteins in the cells in medium and high doses of GAA groups were significantly decreased(P<0.05).Conclusion:Medium and high doses of GAA can inhibit the biological activities of proliferation and migration of the PC9 cells,reduce the glycolysis,and its mechanism may be related to the inhibition of the expressions of the key rate-limiting enzymes HK2 and PKM2.关键词
灵芝酸A/癌,非小细胞肺/己糖激酶2/M2型丙酮酸激酶/糖酵解Key words
Ganoderic acid A/Cancer,non-small cell lung/Hexokinase 2/Pyruvate kinase M2/Glycolysis分类
医药卫生引用本文复制引用
任爱华,董彦伯,苗润芝,吕俞娇,刘岩峰..灵芝酸A对非小细胞肺癌PC9细胞糖酵解及其关键限速酶的影响[J].吉林大学学报（医学版）,2024,50(3):682-688,7.基金项目
吉林省教育厅科学技术研究项目(JJKH20220069KJ,JJKH20180355KJ) （JJKH20220069KJ,JJKH20180355KJ）
北华大学省级大学生创新创业训练计划项目(202310201135) （202310201135）
