title: [Effects of CD147 gene silencing on protein expression of ANXA2, MMP-2 and TIMP-2 by thyroid medullary carcinoma TT cells and biologic characteristics].
abstract: OBJECTIVE
  To study the influence of CD147 gene silencing on the expression of ANXA2, MMP-2 and TIMP-2 of thyroid medullary carcinoma TT cells and related biological characteristics.
  
  
  METHODS
  Protein expression of CD147, ANXA2, MMP-2 and TIMP-2 was detected by immunocytochemistry.RNAi technology was used to identify the specific siRNA sequences and the optimal time point of effective inhibition of CD147 gene. The expression of ANXA2, MMP-2 and TIMP-2 at mRNA and protein levels was detected with RT-PCR and Western blot, respectively. MTT method was used to detect the proliferation of the TT cells, flow cytometry (FCM) to detect the cell cycle and apoptosis changes of TT cells and transwell chamber assays to document the influence of CD147 gene silencing on migration and invasion of the TT cells.
  
  
  RESULTS
  The protein expression of CD147, ANXA2, MMP-2 and TIMP-2 proteins was variable in the TT cells. Two siRNA sequences were identified to effectively silence CD147 gene in the TT cells, in which relative expression of MMP-2 was reduced at both mRNA and protein levels; although the expression of ANXA2 mRNA and protein did not change significantly. TIMP-2 protein expression markedly decreased in an absence of its mRNA expression. The proliferation of the TT cells was inhibited upon the CD147 gene silencing along with a significant increase of G(0)/G(1) phase cells and a decrease of G(2)/M phase cells.However, the proportion of the apoptotic cells in all experimental groups did not change. The number of the penetrating cells through the membrane filters did not show significant changes in all experimental groups in the Transwell chamber assays.
  
  
  CONCLUSIONS
  Through RNAi technology, two CD147 siRNA sequences are identified and shown to effectively inhibit CD147 gene expression of the TT cells. CD147 gene silencing leads to growth inhibition of the TT cells and alteration of the cell cycle. However, silencing CD147 does not significantly affect the apoptosis, migration and invasion of the TT cells.
authors.affiliation: Department of Pathology, the First Affiliated Hospital, Zhengzhou University & Henan Key Laboratory for Tumor Pathology, Zhengzhou 450052, China.
authors.forename: Jiangwei
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Pathology, the First Affiliated Hospital, Zhengzhou University & Henan Key Laboratory for Tumor Pathology, Zhengzhou 450052, China. E-mail:lidaoming@zzu.edu.cn.
authors.forename: Daoming
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Pathology, the First Affiliated Hospital, Zhengzhou University & Henan Key Laboratory for Tumor Pathology, Zhengzhou 450052, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Pathology, the First Affiliated Hospital, Zhengzhou University & Henan Key Laboratory for Tumor Pathology, Zhengzhou 450052, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Huang
authors.affiliation: Department of Pathology, the First Affiliated Hospital, Zhengzhou University & Henan Key Laboratory for Tumor Pathology, Zhengzhou 450052, China.
authors.forename: Zhenqian
authors.initials: Z
authors.lastname: Li
mesh_terms: D017306:Annexin A2 / Q000235:genetics / Q000378:metabolism*; D051926:Basigin / Q000235:genetics* / Q000378:metabolism; D018276:Carcinoma, Medullary / Q000378:metabolism* / Q000473:pathology; D002453:Cell Cycle; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D013964:Thyroid Neoplasms / Q000378:metabolism* / Q000473:pathology; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: C472822:ANXA2 protein, human; D017306:Annexin A2; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C560823:TIMP2 protein, human; D019716:Tissue Inhibitor of Metalloproteinase-2; D051926:Basigin; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2