miR-199a-5p suppresses human bladder cancer cell metastasis by targeting CCR7.
BACKGROUND
C-C chemokine receptor type 7 (CCR7) overexpression correlated with lymphatic metastasis and poor prognosis is a major obstacle to bladder cancer treatment. Recent studies have revealed that miR-199a-5p was significantly abnormal expressed in several solid tumors and functioned as oncogene or tumor suppressor. This study was aimed to further investigate the effects of miR-199a-5p on the cell metastasis mediated by CCR7 in bladder cancer.


METHODS
Quantitative Real Time PCR (qRT-PCR) was firstly performed to identified the expression of miR-199a-5p and CCR7 in human bladder cancer samples and cell lines. Following that, the effects of miR-199a-5p on cell migratory and invasive activities were assessed by wound healing and Matrigel invasion assays, respectively. Finally, luciferase reporter assay and western blot were employed to investigate whether CCR7 could directly interact with miR-199a-5p.


RESULTS
miR-199a-5p downregulation and CCR7 upregulation were firstly observed in bladder cancer samples and cell lines. In addition, both miR-199a-5p downregulation and CCR7 upregulation were significantly involved in bladder cancer clinicopathological features. Moreover, overexpression of miR-199a-5p could inhibit baldder cancer cell migration and invasion. miR-199a-5p was confirmed to be able to target the 3' untranslated region (UTR) of CCR7 and regulate the expression of CCR7, Matrix metalloproteinases 9 (MMP-9) and Epithelial-Mesenchymal Transition (EMT)-related proteins.


CONCLUSIONS
Our findings added newer insights into the multifaceted role played by miR-199a-5p/CCR7 in bladder cancer, prompting for the first time this miRNA/chemokine axis that regulates cell metastasis. The results strongly supported miR-199a-5p as a potential therapeutic agent and diagnostic marker of bladder cancer.
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China.
Mi
M
Zhou
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China.
Shuai
S
Wang
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China.
Linyi
L
Hu
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China.
Feng
F
Liu
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China. kaiguisongyang@163.com.
Qi
Q
Zhang
Department of Urology, Zhejiang Provincial People's Hospital, 158 Shangtang Road, Hangzhou, Zhejiang Province, 310014, People's Republic of China. kiti006@163.com.
Dahong
D
Zhang
D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000502:physiology*; D008875:Middle Aged; D009362:Neoplasm Metastasis; D054400:Receptors, CCR7 / Q000096:biosynthesis*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology*
C515370:CCR7 protein, human; D035683:MicroRNAs; D054400:Receptors, CCR7; C531408:mirn199 microRNA, human
Bladder cancer; CCR7; Metastasis; miR-199a-5p