title: Effect of the interaction between MiR-200b-3p and DNMT3A on cartilage cells of osteoarthritis patients.
abstract: The aim of this research is to explore the effect of miR-200b-3p targeting DNMT3A on the proliferation and apoptosis of osteoarthritis (OA) cartilage cells. Quantitative RT-PCR was performed to analyse the expression of miR-200b-3p, DNMT3A, MMP1, MMP3, MMP9, MMP13 and COL II in normal and OA cartilage tissues. The dual-luciferase reporter assay and Western blot assay were conducted to confirm the targeting relationship between miR-200b-3p and DNMT3A. We also constructed eukaryotic expression vector to overexpress miR-200b-3p and DNMT3A. We detected the expression level of MMPs and COL II in stable transfected cartilage cells using RT-PCR and Western blot. Cell proliferation and apoptosis were evaluated using the MTS, pellet culture and Hoechst 33342 staining method. Finally, we explored the effect of miR-200b-3p targeting DNMT3A on the proliferation and apoptosis of OA cartilage cells. The results of RT-PCR indicated that both miR-200b-3p and COL II were down-regulated in OA cartilage tissues, while the expression of DNMT3A and MMPs was up-regulated in OA cartilage tissues. The expressions of DNMT3A, MMPs and COL II detected by Western blot showed the same trend of the results of RT-PCR. The dual-luciferase reporter assay and Western blot assay confirmed the targeting relationship between miR-200b-3p and DNMT3A. In overexpressed miR-200b-3p cartilage cells, DNMT3A and MMPs were significantly down-regulated, COL II was significantly up-regulated, cell viability was enhanced and apoptosis rate was decreased (P < 0.05). In overexpressed DNM3T cartilage cells, MMPs were significantly up-regulated, COL II was significantly down-regulated, cell viability was weakened and apoptosis rate was increased (P < 0.05). MiR-200b-3p inhibited the secretion of MMPs, promoted the synthesis of COL II and enhanced the growth and proliferation of OA cartilage cells through inhibiting the expression of DNMT3A.
authors.affiliation: Department of Laboratory Medicine, The First People's Hospital of Yancheng City, Yancheng, Jiangsu, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.affiliation: Yancheng TCM Hospital Affiliated To Nanjing University of Chinese Medicine, Yancheng, Jiangsu, China.
authors.forename: Yunjuan
authors.initials: Y
authors.lastname: Tao
authors.affiliation: Clinical Medicine School, Ningxia Medical University, Yinchuan, Ningxia, China.|Department of Laboratory Medicine, The Sixth People's Hospital of Yancheng City, Yancheng, Jiangsu, China.
authors.forename: Anquan
authors.initials: A
authors.lastname: Shang
authors.affiliation: Department of Laboratory Medicine, The Sixth People's Hospital of Yancheng City, Yancheng, Jiangsu, China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Clinical Medicine School, Ningxia Medical University, Yinchuan, Ningxia, China.
authors.forename: Yujie
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Laboratory Medicine, The First Hospital of Ningbo City, Ningbo, Zhejiang, China.
authors.forename: Liqing
authors.initials: L
authors.lastname: Hu
authors.affiliation: Clinical Medicine School, Ningxia Medical University, Yinchuan, Ningxia, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou, Zhejiang, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Laboratory Medicine, The First People's Hospital of Yancheng City, Yancheng, Jiangsu, China.
authors.forename: Naizhou
authors.initials: N
authors.lastname: Guo
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000328:Adult; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D002356:Cartilage / Q000378:metabolism / Q000473:pathology; D049109:Cell Proliferation / Q000235:genetics; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics* / Q000378:metabolism; D000090205:DNA Methyltransferase 3A; D005786:Gene Expression Regulation; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D008875:Middle Aged; D010003:Osteoarthritis / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
chemical_list: D020413:3' Untranslated Regions; C000717967:DNMT3A protein, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A
keywords: 
  DNMT3A
  ; cartilage cell; miR-200b-3p; osteoarthritis