微小核糖核酸-299-3p藉由調控VEGFA影響上泌尿道上皮癌細胞的增生、爬行

和侵襲能力
王建勝^{1, 2}、徐偉齊³、李怡琛^{4, 5}、李威明^{2, 3, 4, 6}、張玲麗^{4, 7}、黃阿梅^{1, 8}、林慧惠²、吳文正^{2, 3, 4}、
李經家^{2, 3}、柯宏龍^{2, 3, 4, 9*}
高雄醫學大學 臨床醫學研究所 ¹ ; 高雄醫學大學附設醫院 泌尿科 ² ; 高雄醫學大學 醫學系
泌尿學科 ³， 醫學研究所 ⁴，解剖學科 ⁵ ; 衛生福利部屏東醫院泌尿科 ⁶ ;
高雄醫學大學 微生物與免疫學科 ⁷；生物化學科 ⁸ ; 高雄市立大同醫院 泌尿科 ⁹

MiR-299-3p Inhibits Cell Proliferation, Motility and Invasion through VEGFA in Upper Tract Urothelial Carcinoma
Chien-Sheng Wang ^{1, 2}, Wei-Chi Hsu ³, Yi-Chen Lee ^{4, 5}, Wei-Ming Li ^{2, 3, 4, 6}, Lin-Li Chang ^{4, 7},
A-Mei Huang ^{1, 8}, Hui-Hui Lin ², Wen-Jeng Wu ^{2, 3, 4}, Ching-Chia Li ^{2, 3}, Hung-Lung Ke ^{2, 3, 4, 9 *}

¹ Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ²Department of Urology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan. ³ Department of Urology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ⁴ Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ⁵ Department of Anatomy, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ⁶ Department of Urology, Ministry of Health and Welfare Pingtung Hospital, Pingtung, Taiwan. ⁷ Department of Microbiology, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ⁸ Department of Biochemistry, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan. ⁹ Department of Urology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.

Purpose: Upper tract urothelial carcinoma (UTUC) is a rare tumor with extraordinarily different feature between Eastern and Western countries. Vascular endothelial growth factor-A (VEGFA) was originally shown to be secreted signaling protein as well as a regulator of vascular development and cancer progression. MiR-299-3p governs tumor suppressive functions in several tumors. We aimed to elucidate the molecular mechanism underlying the regulation of VEGFA by miR-299-3p in UTUC.
Materials and methods: VEGFA expression was evaluated by immunohistochemistry in one hundred and forty human UTUC tissues. We demonstrated that the regulatory relationship between VEGFA and miR-299-3p by real-time PCR, western blot and dual luciferase reporter assay. The role of the miR-299-3p/VEGFA pathway in proliferation, motility and invasion was analyzed in vitro.

Results: VEGFA high expression is significantly associated to stage (P= 0.005), grade (P= 0.004), distant metastasis (P= 0.003), and cancer death (P= 0.001). High VEGFA expression was correlated with poor progression-free survival (P= 0.001) and cancer-specific survival (P< 0.001). Knock-down VEGFA repressed UTUC cell proliferation, migration and invasion. MiR-299-3p overexpression inhibited VEGFA protein expression by directly targeting its 3'-UTR. The functional studies indicated that VEGFA overexpression reversed the miR-299-3p reduced tumor cell proliferation, migration and invasion.

Conclusion: These findings suggest that miR-299-3p may suppress UTUC cell proliferation, motility and invasion by targeting VEGFA.