Studies on proliferation and migration effects and mechanisms of M3 mAChR in NSCLC cells / 中国药理学通报

ABSTRACT

Aim To investigate the effect of M3 mAChR on the proliferation and migration of NSCLC and the molecular mechanisms. Methods CCK-8 as-say,Wound-healing assay and Transwell invasion as-saywere used to determine the cell proliferation,migra-tion and invasion. Calcium imaging was used to identi-fy the M3 mAChR subtype mediating carbachol-in-duced increase in intracellular calcium. Western blot was used to determine the protein level of proliferation, migration and cell cycle related signaling molecules. Results Following the treatment with M3R antagonists 1. 25 ~ 80 μmol · L - 1 for 48 h,the proliferation of NSCLC cells was inhibited,the inhibitory effectR2-8018 > Darifenacin,H1299 > H460. After the treat-ment with R2-8018,the migration and invasion of H1299 significantly declined. Western blot showed that the protein level of p-Akt,p-GSK3β,cyclinD1 de-clined significantly with the increase of time and that the protein level of p21 increased. Conclusion M3R antagonists induce cell cycle arrest by suppressing the activation of Akt,down-regulating GSK3β and cy-clinD1,and up-regulating p21,then inhibiting the proliferation of NSCLC cells. It also inhibits the inva-sion and migration by down-regulating MMP-2.