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19 January 2020 : Animal Research  

Mitofusin-2 (Mfn-2) Might Have Anti-Cancer Effect through Interaction with Transcriptional Factor SP1 and Consequent Regulation on Phosphatidylinositol Transfer Protein 3 (PITPNM3) Expression

Tao Tang1AE*, Xuxiong Tao1ABC, Xing Bao1BC, Jun Chen1BC, Jingyu Dai1CD, Jinjun Ye1BDF, Yukuang Yan1CF

DOI: 10.12659/MSM.918599

Med Sci Monit 2020; 26:e918599

Abstract

BACKGROUND: The aim of this study was to explore the influence of mitofusin-2 (Mfn-2) on phosphatidylinositol transfer protein 3 (PITPNM3) and tumor growth and the potential mechanism behind the regulation of Mfn-2 on PITPNM3 in hepatic carcinoma cell line SMMC-7721.

MATERIAL AND METHODS: We obtained promoter sequence of PITPNM3 gene from University of Santa Cruz (UCSC) genomic database, and we predict transcriptional factor of PITPNM3 genes by JASPAR database. Target transcription factor was determined by comparison of binding sites number for promoter. SMMC-7721 cells were transfected with expression plasmid containing Mfn-2, transcription factor gene and PITPNM3. The cells transfected with empty vector were used as control. Real-time polymerase chain reaction was used to determine the mRNA level of target genes. Co-immunoprecipitation (Co-IP) assay was used to determine the interaction between Mfn-2 and target transcription factor. Chromatin immunoprecipitation assay (ChIP) assay was used to determine the binding of transcription factor with PITPNM3 promoter. Tumorigenicity assay was used to compare the effect of Mfn-2, SP1, and PITPNM3 on tumor development.

RESULTS: SP1 was selected as the target transcriptional factor. In the Co-IP assay, Mfn-2 was shown to interact with SP1. In the ChIP assay Mfn-2 transfection resulted in decreased binding number of SP1 with PITPNM3 promoter. Furthermore, PITPNM3 mRNA levels were significantly increased in SMMC-7721 cells transfected with SP1 but were decreased after transfection with Mfn-2. In nude mice, PITPNM3 and SP1 upregulation lead to larger tumor lump and conversely Mfn-2 upregulation lead to smaller tumor lump.

CONCLUSIONS: Mfn-2 could suppress expression of PITPNM3 through interaction with transcription factor SP1; Mfn-2 may have anti-tumor activity; SP1 and PITPNM3 may promote tumor development.

Keywords: Carcinoma, Acinar Cell, Caspase 1, Drug Screening Assays, Antitumor, Binding Sites, CCAAT-Enhancer-Binding Protein-beta, Calcium-Binding Proteins, carcinogenesis, GTP Phosphohydrolases, Membrane Proteins, Mitochondrial Proteins, Promoter Regions, Genetic, Protein Binding, RNA, Messenger, Sp1 Transcription Factor

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750