title: Downregulation of rho-associated protein kinase 1 by miR-124 in colorectal cancer.
abstract: OBJECTIVE
  To investigate the roles and interactions of rho-associated protein kinase (ROCK)1 and miR-124 in human colorectal cancer (CRC).
  
  
  METHODS
  Expression of ROCK1 protein was examined by Western blotting, and quantitative reverse transcriptase PCR was performed to measure expression of ROCK1 mRNA and miR-124. Two cancer cell lines were transfected with pre-miR-124 (mimic) and anti-miR-124 (inhibitor) and the effects on ROCK1 protein and mRNA expression were observed. In addition, cell proliferation was assessed via a 5-ethynyl-2' deoxyuridine assay. Soft agar formation assay, and cell migration and invasion assays were used to determine the effect of survivin on the transformation and invasion activity of CRC cells.
  
  
  RESULTS
  miR-124 was significantly downregulated in CRC compared to normal specimens (0.603 ± 0.092 vs 1.147 ± 0.286, P = 0.016) and in metastatic compared to nonmetastatic CRC specimens (0.416 ± 0.047 vs 0.696 ± 0.089, P = 0.020). Expression of miR-124 was significantly associated with CRC metastasis, tumor T and N stages, and tumor grade (all P < 0.05). ROCK1 protein was significantly increased in CRC compared to normal tissues (1.896 ± 0.258 vs 0.866 ± 0.136, P = 0.026), whereas ROCK1 mRNA expression was unaltered (2.613 ± 0.251 vs 2.325 ± 0.246). miR-124 and ROCK1 were inversely expressed in CRC tissues and cell lines. ROCK1 mRNA was unaltered in cells transfected with miR-124 mimic and miR-124 inhibitor, compared to normal controls. There was a significant reduction in ROCK1 protein in cells transfected with miR-124 mimic and a significant increase in cells transfected with miR-124 inhibitor (Ps < 0.05). Transformation and invasion of cells transfected with miR-124 inhibitor were significantly increased compared to those in normal controls (P < 0.05). Cells transfected with miR-124 inhibitor showed increased cell proliferation.
  
  
  CONCLUSIONS
  miR-124 promotes hyperplasia and contributes to invasion of CRC cells, but downregulates ROCK1. ROCK1 and miR-124 may play important roles in CRC.
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Zuo-Wu
authors.initials: ZW
authors.lastname: Xi
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Shi-Yong
authors.initials: SY
authors.lastname: Xin
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Li-Qing
authors.initials: LQ
authors.lastname: Zhou
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Hai-Xin
authors.initials: HX
authors.lastname: Yuan
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Zuo-Wu Xi, Kai-Xuan Chen, Department of Anorectal Surgery, Henan province Hospital of Traditional Chinese Medicine (Second Affiliated Hospital of Henan University of traditional Chinese medicine), Zhengzhou 450002, Henan Province, China.
authors.forename: Kai-Xuan
authors.initials: KX
authors.lastname: Chen
mesh_terms: D002465:Cell Movement; D049109:Cell Proliferation; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D015536:Down-Regulation; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D045325:HCT116 Cells; D019073:HT29 Cells; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D009367:Neoplasm Staging; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D054460:rho-Associated Kinases / Q000235:genetics / Q000378:metabolism*
chemical_list: C509207:MIRN124 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases
keywords: Cell invasion; Colorectal cancer; Rho-associated protein kinase; miR-124