KRAS促进肺癌细胞糖酵解  

Homo sapiensv-Ki-ras2 Kirsten rat sarcoma viral oncogenes homologue promotes glycolysis in lung cancer cells

作　　者：李晓静[1] 唐青[1] 胡义德[1] 

机构地区：[1]第三军医大学新桥医院肿瘤科,重庆400037

出　　处：《第三军医大学学报》2015年第11期1102-1107,共6页Journal of Third Military Medical University

摘　　要：目的探讨KRAS对肺癌细胞糖酵解的影响及其初步机制。方法采用Real-time PCR和Western blot检测KRAS、HK2、PKM2在肺癌细胞系H1299、A549、SPC-A1中的表达水平,应用干化学方法检测肺癌细胞培养48 h后培养基上清中乳酸的浓度。采用RNA干扰技术,将KRAS-shRNA感染H1299细胞,实验分为3组:空白对照组(CON组)、阴性对照组(NC组)、慢病毒干扰组(KD组)。采用Real-time PCR和Western blot检测KRAS敲低效率及敲低KRAS基因前后H1299细胞中HK2、PKM2的表达变化。将TNF-α作用于NC组和KD组细胞,检测KRAS、HK2、PKM2的表达水平及乳酸浓度的变化。结果在3株肺癌细胞系中,H1299细胞中KRAS和HK2、PKM2的表达水平最高,乳酸水平也高于A549、SPC-A1细胞。与空白对照组相比,KRAS-shRNA感染H1299细胞后,KRAS基因拷贝以及蛋白表达水平被显著抑制(P<0.05),HK2、PKM2基因拷贝及蛋白表达水平显著下降,培养基中乳酸浓度明显降低(P<0.05)。以TNF-α作用NC组、KD组细胞后,KRAS、HK2、PKM2基因和蛋白表达均升高(P<0.05)。KRAS-shRNA可部分抑制TNF-α升高的乳酸浓度(P<0.05)。结论 KRAS能通过调节HK2和PKM2促进肺癌细胞的糖酵解。Objective To determine the effect of homo sapiensv-Ki-ras2 Kirsten rat sarcoma viral oneogenes homologue （KRAS） on the glycolysis in lung cancer ceils and investigate the underlying possible mechanism. Methods The expression of KRAS, hexokinase 2 （HK2） , and pyruvate kinase M2 （PKM2） in 3 lung cancer cell lines H1299, A549 and SPC-A was determined with real-time PCR and Western blotting. Lactate concentration was assessed in 48 hours＇ culture. After H1299 cells were transfeeted with negative control and KRAS-shRNA respectively, the expression of the 3 molecules was assessed with real-time PCR and Western blot analysis. After treatment with TNF- alpha in the negative control and H1299-KRAS-shRNA cells, the expression levels of HK2 and PKM2, as well as the lactate concentration were assessed. Results The expression of KRAS, HK2 and PKM2 as well as the lactate secretion in H1299 cells were the highest among above 3 lung cancer cell lines. Compared to the negative control group, lentiviral shRNA interference significantly inhibited KRAS expression at mRNA and protein levels （ P 〈 0.05 ）. Moreover, the mRNA and protein levels of HK2 and PKM2 were also decreased in KRAS-shRNA transfected H1299 cells, compared with negative control （P 〈 0.05 ）. TNF-α enhanced KRAS, HK2 and PKM2 in both the negative control and H1299-KRAS-shRNA cells （P 〈 0.05 ）. In addition, KRAS-shRNA significantly decreased TNF-α elevated lactate concentrations in the supernatant compared with the negative control （P 〈 0.05 ）. Conclusion KRASpromotes the glycolysis in lung cancer cells via modulating glycolysis related enzymes.

关 键 词：KRAS 肺癌 糖酵解 SHRNA 

分 类 号：Q591.4[生物学—生物化学] R734.2[医药卫生—肿瘤]