title: MiR-20a inhibits cutaneous squamous cell carcinoma metastasis and proliferation by directly targeting LIMK1.
abstract: BACKGROUND
  MicroRNA-20a (miR-20a) plays a key role in tumorigenesis and progression. But its function is reverse in different kinds of malignant tumor, and its role and mechanism in cutaneous squamous cell carcinoma (CSCC) remains unclear.
  
  
  OBJECTIVE
  To determine the miR-20a's roles in CSCC and confirm whether LIMK1 is a direct target gene of miR-20a.
  
  
  METHODS
  First miR-20a and LIMK1 expression levels were detected in six pairs of CSCC tissues and corresponding normal skin by qRT-PCR. Then MTT assays and colony formation assays were performed to evaluate the impact of miR-20a on cell proliferation. In addition, scratch migration assays and transwell invasion assays were performed to check miR-20a's effect on cell metastasis. Since LIMK1 (LIM kinase-1) was predicted as a target gene of miR-20a, the changes of LIMK1 protein and mRNA were measured by western blot and qRT-RCR methods after miR-20a overexpression. Moreover the dual reporter gene assay was performed to confirm whether LIMK1 is a direct target gene of miR-20a. Finally LIMK1 mRNA and miR-20a in other 30 cases of CSCC pathological specimens were determined and a correlation analysis was evaluated.
  
  
  RESULTS
  The miR-20a significantly low-expressed in CSCC tissues compared with that in matched normal tissues while LIMK1 has a relative higher expression. MiR-20a inhibited A431 and SCL-1 proliferation and metastasis. Both of LIMK1 protein and mRNA levels were downregulated after miR-20a overexpression. The dual reporter gene assays revealed that LIMK1 is a direct target gene of miR-20a. Furthermore, qRT-PCR results of LIMK1 mRNA and miR-20a in 30 cases of CSCC pathological specimens showed miR-20a is inversely correlated with LIMK1 expression.
  
  
  CONCLUSIONS
  Our study demonstrated that miR-20a is involved in the tumor inhibition of CSCC by directly targeting LIMK1 gene. This finding provides potential novel strategies for therapeutic interventions of CSCC.
authors.affiliation: Department of Plastic and Reconstructive Surgery; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Jianda
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Department of Plastic and Reconstructive Surgery; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Rui
authors.initials: R
authors.lastname: Liu
authors.affiliation: Department of Plastic and Reconstructive Surgery; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Chengqun
authors.initials: C
authors.lastname: Luo
authors.affiliation: Department of Oncoplastic and Reconstructive Surgery; The Affiliated Tumor Hospital of Xiangya Medical School; Changsha City, Hunan, PR China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Medical Genetics; Changsha City, Hunan, PR China.
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.affiliation: Department of Dermatology; Xiangya Hospital; Changsha City, Hunan, PR China.
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.affiliation: Cancer Research Institute; Key Laboratory of Carcinogenesis of Ministry of Health; Changsha City, Hunan, PR China.
authors.forename: Ming
authors.initials: M
authors.lastname: Zhou
authors.affiliation: Department of Pathology; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Zou
authors.affiliation: Department of Oncology; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Peiguo
authors.initials: P
authors.lastname: Cao
authors.affiliation: Department of Oncology; Third Xiangya Hospital; Central South University; Changsha City, Hunan, PR China.
authors.forename: Ke
authors.initials: K
authors.lastname: Cao
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000473:pathology* / Q000556:secondary; D002465:Cell Movement; D049109:Cell Proliferation; D005260:Female; D006801:Humans; D054461:Lim Kinases / Q000235:genetics / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012878:Skin Neoplasms / Q000473:pathology*; D014407:Tumor Cells, Cultured
chemical_list: C534451:MIRN20a microRNA, human; D035683:MicroRNAs; C515892:LIMK1 protein, human; D054461:Lim Kinases
keywords: LIMK1; cutaneous squamous cell carcinoma; metastasis; miR-20a; proliferation