TRPM7 Affects Proliferation and Apoptosis of Human Cerebral Vascular Adventitial Fibroblasts via PI3K/AKT Pathway
GUO Zeming1*, CAI Qiuhong2
The abnormal proliferation of HBVAFs (human cerebral vascular adventitial fibroblasts) contributes to the development of vascular proliferative diseases. This study aimed to investigate whether TRPM7 (transient receptor potential melastatin 7) could affect the proliferation and apoptosis of HBVAFs through PI3K/AKT signaling pathway. The cultured HBVAFs were randomly divided into the following groups: parental (normal cultured HBVAFs), si-NC, si-TRPM7, si-NC+IGF-1 (an activator of PI3K/AKT pathway), si-TRPM7+IGF-1. qRT-PCR was used for determining TRPM7 mRNA expression. The cell proliferation was detected by CCK-8. Flow cytometry was performed to assess cell cycle and apoptosis. The levels of target proteins were evaluated by Western blot. The results showed that si-TRPM7 transfection significantly reduced the mRNA and protein expression of TRPM7 in HBVAFs (P<0.001). Compared with si-NC group, si-TRPM7 transfection reduced cell viability (P<0.001), increased percentage in G0-G1 phase (P<0.001), decreased percentage in S phase (P<0.001), enhanced protein levels of CCND1, CDK2, CDK4 (P<0.001), raised apoptotic rate (P<0.001), reduced protein levels of Bcl-2, p-PI3K, and p-AKT (P<0.001), and elevated protein levels of Bax, cleaved caspase-3 and Cytochrome c (P<0.001). However, these si-TRPM7-mediated changes could be effectively reversed by IGF-1 treatment (P<0.001). These findings indicated that interference of TRPM7 could inhibit the proliferation and induce apoptosis of HBVAFs via inactivation of PI3K/AKT signaling pathway, which provided theoretical basis for TRPM7 serving as a potential therapeutic target for vascular proliferative disease.
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