Junhong Cai1, Ying Zhang2, Sizhe Huang2, Mengdan Yan3, Jingjie Li3, Tianbo Jin3,4, Shan Bao2. 1. Central Laboratory, Hainan General Hospital Haikou, Hainan, China. 2. Department of Gynecology and Obstetrics, Hainan General Hospital Haikou, Hainan, China. 3. Key Laboratory of Resource Biology and Biotechnology in Western China (Northwest University), Ministry of Education, Northwest University Xi'an, Shaanxi, China. 4. School of Life Sciences, Northwest University Xi'an, Shaanxi, China.
Abstract
OBJECTIVE: The study aimed to explore the association between three miRNAs (miR-100-5p, miR-199a-3p and miR-199b-5p) and mTOR induced autophagy in EEC cells. The expression of three miRNAs and autophagy-related genes Beclin1 and LC3 in Ishikawa and KLE cells were detected. METHODS: The effects of three miRNAs on proliferation and apoptosis in EEC cells were analyzed in KLE and ISK cells transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics and inhibitors. Quantitative real-time polymerase chain reaction (qRT-PCR), CCK-8 method, flow cytometry and luciferase reporter assay were used to assess the effects of three miRNAs on cell viability, proliferation, apoptosis, and autophagy. RESULTS: We found an increased expression of Beclin1 and LC3 in Ishikawa and KLE cells transfected by miR-100-5p, miR-199a-3p and miR-199b-5p mimics compared with NC (P<0.05). Additionally, Ishikawa and KLE cells transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics grew more slowly than mock and mimics control (P<0.05); we also found an increased apoptosis incidence of Ishikawa cell transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics (P<0.05). Finally, luciferase reporter results showed miR-100-5p, miR-199a-3p and miR-199b-5p were all down-regulated the luciferase activity in cells transfected with miRNA mimics compared with mock. All results suggested that miR-100-5p, miR-199a-3p and miR-199b-5p may induce the autophagic death of EEC cell through targeting mTOR. CONCLUSIONS: Our results suggested that miR-100-5p, miR-199a-3p and miR-199b-5p may induce the autophagic death of EEC cell through targeting mTOR. IJCEP
OBJECTIVE: The study aimed to explore the association between three miRNAs (miR-100-5p, miR-199a-3p and miR-199b-5p) and mTOR induced autophagy in EEC cells. The expression of three miRNAs and autophagy-related genes Beclin1 and LC3 in Ishikawa and KLE cells were detected. METHODS: The effects of three miRNAs on proliferation and apoptosis in EEC cells were analyzed in KLE and ISK cells transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics and inhibitors. Quantitative real-time polymerase chain reaction (qRT-PCR), CCK-8 method, flow cytometry and luciferase reporter assay were used to assess the effects of three miRNAs on cell viability, proliferation, apoptosis, and autophagy. RESULTS: We found an increased expression of Beclin1 and LC3 in Ishikawa and KLE cells transfected by miR-100-5p, miR-199a-3p and miR-199b-5p mimics compared with NC (P<0.05). Additionally, Ishikawa and KLE cells transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics grew more slowly than mock and mimics control (P<0.05); we also found an increased apoptosis incidence of Ishikawa cell transfected with miR-100-5p, miR-199a-3p and miR-199b-5p mimics (P<0.05). Finally, luciferase reporter results showed miR-100-5p, miR-199a-3p and miR-199b-5p were all down-regulated the luciferase activity in cells transfected with miRNA mimics compared with mock. All results suggested that miR-100-5p, miR-199a-3p and miR-199b-5p may induce the autophagic death of EEC cell through targeting mTOR. CONCLUSIONS: Our results suggested that miR-100-5p, miR-199a-3p and miR-199b-5p may induce the autophagic death of EEC cell through targeting mTOR. IJCEP
Authors: Delphine Garnier; Edward Ratcliffe; Joséphine Briand; Pierre-François Cartron; Lisa Oliver; François M Vallette Journal: Biomedicines Date: 2022-01-06