Abstract:

Objective To explore the effects and mechanism of miR ⁃ 330⁃3p on the proliferation and apoptosis of gastric cancer cells，and aerobic glycolysis process by targeting pyruvate kinase M2（PKM2）gene. Methods The expression levels of SGC⁃7901 in gastric cancer cells and GES⁃1 miR⁃miR⁃330⁃3p in gastric muco⁃ sal epithelial cells were detected by reverse transcription⁃polymerase chain reaction（RT⁃PCR）. The miR⁃NC，miR⁃ 330⁃3p⁃inhibitor，miR⁃330⁃3p⁃mimcs，miR⁃330⁃3p⁃inhibitor+PKM2⁃NC，miR⁃330⁃3p⁃inhibitor+PKM2⁃shRNA， miR⁃330⁃3p⁃mimcs+LV⁃NC and miR⁃330⁃3p⁃mimcs+LV⁃PKM2 were transfected into gastric cancer cells. The proliferation，invasion and apoptosis of cells were detected by CCK8，Transwell assay and Annexin V/PI method. The glycolysis indexes such as glucose utilization rate，production of lactic acid，activities of hexokinase（HK） and lactate dehydrogenase（LDH）were detected. The expression of PKM2，GLUT1，HK2，Bax and Caspase ⁃ 3 proteins was detected by Western Blot. The targeted relationship between miR⁃330⁃3p and PKM2 was detected by double luciferase reporter gene assay. Results （1）The expression level of miR⁃330⁃3p in gastric cancer cell line was higher than that in gastric mucosal epithelial cells，while expression level of PKM2 was lower（P < 0.05）.（2） reporter gene assay showed that compared with that in miR⁃NC group，activity of PKM2 wild⁃type luciferase was decreased in miR⁃330⁃3p group（P < 0.05）.（3）Compared with those in miR⁃NC group，absorbance and number of invasion cells were increased in miR⁃330⁃3p⁃inhibitor group；number of apoptosis cells was decreased；cellular glucose consumption，production of lactic acid，HK and LDH activities were decreased；expression of PKM2， GLUT1 and HK2 was increased，and expression level of pro⁃apoptosis proteins（Bax and Caspase⁃3）was decreased （P < 0.05）. The increase or decrease amplitude in miR⁃330⁃3p⁃inhibitor+PKM2⁃shRNA group was lower than that in miR⁃330⁃3p⁃inhibitor group（P < 0.05）.（4）Compared with that in miR⁃NC group，expression level of miR⁃ 330⁃3p was increased in miR⁃330⁃3p⁃mimcs group（P < 0.05），absorbance and number of invasion cells were decreased，number of apoptosis cells was increased，cellular glucose consumption，production of lactic acid，HK and LDH activities were increased，expression of PKM2，GLUT1 and HK2 was decreased，and expression levels of pro⁃apoptosis proteins（Bax，Caspase⁃3）were increased（P < 0.05）. The increase or decrease amplitude in miR⁃ 330⁃3p⁃mimcs+LV⁃PKM2 group was lower than that in miR⁃330⁃3p⁃mimcs group（P < 0.05）. Conclusion The expression of miR⁃330⁃3p is related to proliferation and apoptosis of gastric cancer cells，and aerobic glycolysis pro⁃ cess，

Key words:

miR ? 330 ? 3p, pyruvate kinase M2 type, gastric cancer, glycolysis, proliferation, apoptosis