miR-30a-3p/MMP2機制調控膀胱癌細胞侵犯與腫瘤增殖的探討
仇光宇1 、張安辰2、陳栢均2、邱培承2、蔡德甫1、陳宏恩1、林宜佳1、林致凡2、黃一勝1,2,3,4
1新光醫院 外科部 泌尿科、2新光醫院 中央研究室、3台北醫學大學 醫學院、4輔仁大學 醫學院
The mechanism of miR-30a-3p/MMP2 system regulates cell invasion and tumor growth in baldder cancer
Kuang-Yu Chou1, An-Chen Chang2, Po-Chun Chen2, Pei-Cheng Ciou 2, Te-Fu Tsai 1, Hung-En Chen1, Yi-Chia Lin1 , Ji-Fan Lin2 and Thomas I-Sheng Hwang1,2,3,4
Department of Urology, Shin Kong Wu Ho-Su Memorial Hospital1, Central Laboratory, Shin Kong Wu Ho-Su Memorial Hospital2, Department of Urology, Taipei Medical University3, Division of Urology, School of Medicine, Fu-Jen Catholic University4, Taipei, Taiwan.
Background:
Bladder cancer (BC) is one of the most common noncutaneous malignancies with a high recurrence rate. Numerous study suggests that matrix metalloproteinases (MMPs) are involved in cancer initiation, invasion, and metastasis. The function of miR-30a acts as a tumor suppressor gene is related to its roles in autophagy and metastasis repression. It is a known fact that miR-30a was down-regulated in the progression of human BC cells. This study aims to investigate the role of miR-30a-5p and -3p (two isoforms of miR-30a) in cancer invasion, by targeting MMPs in BC.
Materials and Methods:
The levels of miR-30a-5p and -3p expression in BC specimens were evaluated from the online database GEPIA. To evaluate the regulatory roles of miR-30a-5p and -3p, the BC cell lines T24, and 5637 were transfected with miR-30a-5p or -3p mimic or negative control microRNA mimic, followed by evaluating cell migratory and invasion activity. The BC cells which stably expressed pre-miR-30a was established to confirm the role of miR-30a in the regulation of cell invasion. Finally, the in vivo orthotopic model was performed to evaluate the effect of miR-30a-3p in the treatment of bladder cancer.
Results:
The expression level of miR-30a-3p, but not miR-30a-5p, was significantly decreased in patients with bladder cancer compared with normal tissue. In in vitro analysis, we also found that the cell migration and invasion activity was inhibited by a miR-30a-3p mimic. However, the miR-30a-5p mimic has no such effect. Moreover, analysis of the miRNA target prediction program confirmed that miR-30a-3p directly targets the 3’-UTR region of MMP2. Transfected the cells with miR-30a-3p mimic reduced MMP2 protein and mRNA expression. Finally, the in vivo orthotopic model indicated that overexpression of miR-30a-3p diminished tumor growth. The H&E staining revealed that bladder cancer cells invaded surrounding muscle tissue and their attenuation by miR-30a-3p overexpression.
Conclusions:
Our results demonstrate that miR-30a-3p has an obvious effect on the suppression of BC invasion by directly suppressing MMP2 expression. The microRNA replenishment therapy by delivery of miR-30a-3p into BC cells represents a promising novel therapeutic strategy.