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KRAS促进肺癌细胞糖酵解

, PP. 1102-1107

Keywords: KRAS,肺癌,糖酵解,shRNA

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Abstract:

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-timePCR和Westernblot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48h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-timePCR和Westernblot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。

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