摘要
目的探讨Dis312基因敲除对胃癌细胞葡萄糖代谢的影响，并初步探讨该作用的分子机制。方法利用CRISPR／Cas9技术敲除胃癌细胞Dis312基因，然后利用生化法检测细胞培养液乳酸含量，ELISA法检测细胞内磷酸果糖激酶（PFK）含量，蛋白质印迹法检测哺乳动物雷帕霉素靶蛋白（mTOR）、缺氧诱导因子-1α（HIF-1α）、葡萄糖转运蛋白1（GLUT-1）、丙酮酸激酶M2（PKM2）蛋白表达水平。结果胃癌细胞Dis312基因敲除后，细胞培养液乳酸含量和细胞内PFK含量均升高；mTOR蛋白及葡萄糖代谢相关蛋白HIF-1α、GLUT-1、PKM2水平明显升高。结论胃癌细胞Dis312基因敲除后可以促进胃癌细胞的糖酵解，其作用机制可能是通过roTOR通路上调HIF-1α、GLUT-1、PKM2水平而实现。
Objective To explore the effects of Dis312 knockout on the glucose metabolism of gastric cancer cells. Methods CRISPR/Cas 9 system was used to knock out Dis312 gene. The amount of lactic acid in the culture media was measured by biological methods. The level of phosphate fructose kinase （PFK） in the cells was also detected by ELISA. The levels of mammalian target of rapamycin （mTOR） , hypoxia inducible factor - 1α（ HIF - 1α） , glucose transporter 1 （ GLUT - 1 ） , and pyruvate kinase M2 （PKM2） were detected using Western blotting. Results Knockout of Dis312 in gastric cancer cells resulted in increases in the level of lactic aeid in the culture media and PFK levels in the cells, while up - regulating the expression of mTOR, HIF - 1α, GLUT - 1, and PKM2. Conclusion Knockout of Dis312 promotes the glycolysis of gastric cancer cells, which may be related with the up - regulation of HIF - 1α, GLUT - 1, and PKM2 in the mTOR pathway.
出处
《徐州医科大学学报》
CAS
2017年第12期776-778,共3页
Journal of Xuzhou Medical University
基金
江苏省“六大人才高峰”资助项目（2014-WSW-032）
江苏省“科教强卫工程”青年医学人才项目资助（QN-RC2016790）
