Abstract: Objective To study the effect of long noncoding RNA human microRNA17 cluster host gene (LncRNA MIR17HG) on the malignant biological behavior of bladder cancer (BCa) cells by targeting microRNA (miR)-130a-3p. Methods LncRNA MIR17HG and miR-130a-3p were measured in SV-HUC-1 human urinary tract epithelial cells and human BCa cells (T24, UM-UC-3, 5637) by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The relationship between LncRNA MIR17HG and miR-130a-3p was detected by dual-luciferase reporter assay. BCa T24 cells were divided into Control, sh-NC, sh-LncRNA MIR17HG, sh-LncRNA MIR17HG+NC inhibitor, and sh-LncRNA MIR17HG+ miR-130a-3p inhibitor groups. LncRNA MIR17HG and miR-130a-3p were detected by RT-qPCR. Cell proliferation, migration, invasion, apoptosis, and related proteins in each group were detected by Cell Counting Kit 8, cell scratch, and Transwell assays, flow cytometry, and Western blot, respectively. Results LncRNA MIR17HG was increased and miR-130a-3p was decreased in human BCa cells (P<0. 05), with the greatest difference in T24 cells. T24 cells were therefore selected for subsequent experiments. Wild-type (WT) LncRNA MIR17HG+miR-130a-3p mimic cells showed lower luciferase activity than WT LncRNA MIR17HG+mimicNC cells (P<0. 05). In the Control and sh-NC groups, sh-LncRNA MIR17HG cells showed decreases in LncRNA MIR17HG, survival rate, invasion number, scratch-healing rate, proliferating cell nuclear antigen, and N-cadherin in T24 cells, and increases in miR-130a-3p, apoptosis rate, cleaved caspase-3, and E-cadherin (P<0. 05). In the shLncRNA MIR17HG and sh-LncRNA MIR17HG+NC inhibitor groups, sh-LncRNA MIR17HG+miR-130a-3p inhibitor cells showed no clear changes in LncRNA MIR17HG in T24 cells, but increases in survival rate, invasion number,scratch-healing rate, PCNA, and N-cadherin, and decreases in miR-130a-3p, apoptosis rate, cleaved caspase-3, and E-cadherin ( P<0. 05). Conclusions Knockdown of LncRNA MIR17HG may inhibit the malignant biological behavior of BCa cells by targeting upregulation of miR-130a-3p.