Muscarinic Acetylcholine Receptor M1 Promotes Cell Metastasis and Resists Cell Apoptosis in Prostate Cancer

WANG Qianhui, ZHANG Mi, WANG Hong, CHEN Jinying, HUANG Yanping, XU Chen*

(College of Life Science, Chongqing Medical University, Chongqing 400016, China)

Abstract:

The aim of this study was to investigate the effects of CHRM1 (muscarinic acetylcholine receptor M1) on the proliferation, metastasis and anti-apoptosis of prostate cancer cells by regulating PI3K/AKT signaling pathway. Western blot, immunofluorescence and the other methods were used to detect the expression of CHRM1 in prostate cancer cells. Human prostate cancer cells PC-3, LNCaP, DU145 were cultured in vitro, and the cells were treated by CHRM1 agonist CAR (carbachol) and CHRM1 specific inhibitor PIN (pirenzepine). PC-3 cells were infected with CHRM1 RNAi lentivirus and a stably transfected strain of CHRM1 knockdown was constructed. CCK8 cell proliferation assay, plate cloning assay, cell migration and invasion experiments, flow cytometry detection and observation under transmission electron microscope were used to explore the proliferation, metastasis and apopto-sis levels of CHRM1 in prostate cancer. Western blot was used to detect the expression of epithelial-mesenchymal markers and PI3K/AKT signals in prostate cancer cells knocking down CHRM1. The results showed that CHRM1 expressed abundantly in prostate cancer cells. In our assays, the group treated with CAR promoted the proliferation, colony formation, migration, invasion and anti-apoptosis abilities of prostate cancer, while the group PIN inhibited the proliferation, colony formation, migration, invasion and anti-apoptosis abilities. What’s more, CHRM1 knockdown also inhibited the migration, invasion and anti-apoptosis abilities, and apoptotic bodies appeared under electron microscope. The ability of metastasis was related to EMT (epithelial mesenchymal transformation) in the experiment of cell migration and invasion. Moreover, CHRM1 regulates tumor cell progression through PI3K/AKT signaling pathway. The results of this study suggested that CHRM1 could regulate PI3K/AKT signaling pathway to promote proliferation, metastasis and resist apoptosis in prostate cancer cells.