Bladder cancer (BC) is a familiar malignancy with high morbidity and mortality. The effect of treatment is unsatisfactory after the metastasis and invasion of BC. Hence, more studies should be carried out to explore the metastasis of BC. RT‐qPCR or/and western blot was conducted to evaluate miR‐494‐3p, KLF9, and RGS2 expression. Cell proliferation and invasion were estimated by MTT assay and transwell assay, respectively. Cell migration was tested by wound healing assay and transwell assay. Dual‐luciferase reporter gene assay was employed to validate the interplay between miR‐494‐3p and KLF9 mRNA. The interaction between KLF9 and RGS2 promoter was verified using dual‐luciferase reporter gene assay and chromatin immunoprecipitation (ChIP) assay. miR‐494‐3p expression was upregulated, whereas KLF9 and RGS2 were downregulated in BC cells. miR‐494‐3p inhibition was competent to limit the growth of BC cells. KLF9 knockdown abolished the miR‐494‐3p depletion‐mediated inhibitory growth of BC cells. Mechanistically, we found that KLF9 was a downstream gene of miR‐494‐3p and could bind to the promoter region of RGS2 to promote the expression of RGS2. Moreover, RGS2 knockdown abrogated the suppressive effects of miR‐494‐3p knockdown on the proliferation, migration, and invasion of BC cells. Notably, miR‐494‐3p inhibition obstructed the tumor growth in nude mice. miR‐494‐3p silencing inhibited the progression of BC by regulating the KLF9/RGS2 axis in vitro and in vivo, which laid the foundation for experiments of miR‐494‐3p in BC and provided therapeutic targets for BC.
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