miR-199a-5p regulates the expression of metastasis-associated genes in B16F10 melanoma cells.
MicroRNAs are regulatory factors that play important roles in tumor development, invasion and metastasis. Previously, we showed that miR-199a is abnormally expressed in clinical melanoma specimens and expression was closely associated with clinical features of metastasis. However, the exact molecular mechanisms by which miR-199a-5p influences melanoma invasion and metastasis remains unclear. In this study, we investigated gene expression changes of metastasis-associated genes in B16F10 melanoma cells following targeted silencing or overexpression of miR-199a-5p, using mouse tumor metastasis PCR arrays. Comparison of gene expression changes in miR-199a-5p-silenced versus overexpressing cells identified a set of upregulated genes (> 2-fold) including Cd44, Cdh1, Cxcr4, Etv4, Fxyd5, Rpsa, Mmp3, Myc, Rb1, Tcf20, Hprt1, Actb1 and downregulated genes (> 2-fold) including Ctsk, Itga7 and Tnfsf10. Regulation of a subset of these genes (Myc, Tnfsf10 and Cd44) following miR-199a-5p silencing or overexpression was validated by reverse transcription-polymerase chain reaction (RT-PCR) and western blot. In conclusion, our study demonstrates that miR-199a-5p regulates melanoma metastasis-related genes, and may provide a basis for the development of novel, molecularly targeted drugs.
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Jianda
J
Zhou
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Rui
R
Liu
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Yang
Y
Wang
Institute of Medical Physics and Engineering, Department of Engineering Physics, Tsinghua University Beijing, China.
Jingtian
J
Tang
Cleft Lip and Palate Treatment Center, Second Affiliated Hospital, Shantou University Medical College Shantou, China.
Shijie
S
Tang
Department of Dermatology, Xiangya Hospital, Central South University Changsha, China.
Xiang
X
Chen
State Key Laboratory of Medical Genetics, Central South University Changsha, China.
Kun
K
Xia
Cancer Research Institute, Key Laboratory of Carcinogenesis of Ministry of Health, Central South University Changsha, China.
Wei
W
Xiong
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Dan
D
Xu
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Shaohua
S
Wang
Department of Plastic and Reconstructive Surgery, Third Xiangya Hospital, Central South University Changsha, China.
Quanyong
Q
He
Department of Oncology, Third Xiangya Hospital, Central South University Changsha, China.
Ke
K
Cao
D000818:Animals; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D008546:Melanoma, Experimental / Q000235:genetics* / Q000378:metabolism / Q000556:secondary*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009361:Neoplasm Invasiveness; D020411:Oligonucleotide Array Sequence Analysis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012878:Skin Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D014162:Transfection
D035683:MicroRNAs; C534780:Mirn199 microRNA, mouse
Melanoma; metastasis PCR array; miR-199a-5p