miR-30a-3p藉由標靶細胞自噬相關基因及基質金屬蛋白酶抑制膀胱癌細胞自嗜作用及細胞侵犯
仇光宇1 、陳栢均2、楊尚哲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
miR-30a-3p inhibits bladder cancer autophagy and invasion by suppressing autophagic genes and matrix metalloproteinases expression
Kuang-Yu Chou1, Po-Chun Chen2, Shan-Che Yang2, 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: miR-30a is tumor suppressor genes that were down-regulated in progression of bladder cancer (BC). The tumor-suppressive function of miR-30a is related to its roles in autophagy and metastasis repression. Herein, we investigate role of miR-30a-5p and -3p, two isoforms of miR-30a, in cancer progression by targeting autophagy- and metastasis-related genes in BC.
Materials and Methods: To evaluate the regulatory roles of miR-30a-5p/3p, T24 and 5637 BC cells were transfected with miR-30a-5p/3p mimic or negative control microRNA mimic, followed by evaluating autophagy and cell mobility phenotype. Expression levels of predicted targeting genes that involved in metastasis and autophagy formation were accessed by QPCR and Western blot. Chemosensitivity of BC cells transfected miR-30a-5p/3p mimic was evaluated by cell viability assay, caspase-3 activity and annexin V/PI staining. Cell mobility was examined by migration and invasion assay. Finally, the BC cells which stably expressed pre-miR-30a was established to confirm the role of miR-30a in metastasis and autophagy regulation.
Results: The autophagy activity was suppressed in BC cells introduced with miR-30a-5p and -3p mimic. Suprisingly, transfected with miR-30a-3p showed obvious effect on suppression of autophagy-related genes including including ATG5, ATG12, and Beclin-1. BC cells transfected with miR-30a-3p were found to improve chemosensitivity after cisplatin exposure. Meanwhile, the cell mobility was also dramatically inhibited by miR-30a-3p mimic than miR-30a-5p mimic. The results indicated matrix metalloproteinase (MMP)-2 and MMP-9 expression level were reduced after miR-30a-3p mimic transfection. Finally, the miR-30a-expressing cells showed decreased autophagy and mobility in BC cells.
Conclusions: Our results demonstrate that miR-30a-3p have great effect on suppression of BC progression through inhibiting autophagy and metastasis-related genes. The microRNA replenishment therapy by delivery of miR-30a-3p into BC cells represents a promising novel therapeutic strategy.
Keywords: Autophagy, invasion, ATGs, MMP, bladder cancer, miR-30a.