Qiang Li1, Sugui Wang1, Ziyu Wu1, Yuzhong Liu2. 1. Department of Urology, Huai'an Hospital Affiliated to Xuzhou Medical University and Second People's Hospital of Huai'an, Huai'an, 223000, China. 2. Department of Urology, Traditional Chinese Medical Hospital of Siyang County, 15 Jiefangbei Road, Siyang, 223700, China. Electronic address: yuzhong_liu08@163.com.
Abstract
PURPOSE: We investigated DDX11-AS1 effects on bladder cancer (BLCA) progression to identify a new potential therapeutic target for BLCA. METHODS: BLCA cases (n = 108) were enrolled. SW780 and J82 cells were transfected. Cell counting kit-8 (CCK-8) assay, wound healing assay and transwell migration assay was conducted. Cell cycle and apoptosis was detected by flow cytometry. Luciferase reporter assay was performed. DDX11-AS1, miR-499b-5p and CDK6 mRNA expression in tissues/cells was determined by quantitative real-time polymerase chain reaction (qRT-PCR). In vivo experiment was performed using nude mice. CDK6 and Ki67 proteins expression in cells and xenograft tumors were researched by Western blot and immunohistochemistry. RESULTS: Overexpressed DDX11-AS1 in BLCA was associated with poor outcome of patients. Compared with siCtrl group, SW780 and J82 cells of siDDX11-AS1 group had lower OD450 value (P < 0.01), less cells in S phase, more apoptosis cells (P < 0.05), higher relative wound width (P < 0.05) and less invasive cell number (P < 0.01). DDX11-AS1 promoted CDK6 expression via inhibiting miR-499b-5p. Compared with oe-DDX11-AS1 group, SW780 cells of oe-DDX11-AS1 + miR-499b-5p mimic group and oe-DDX11-AS1 + siCDK6 group had lower OD450 value (P < 0.01), less cells in S phrase, more apoptosis cells (P < 0.01), higher relative wound width (P < 0.05) and less invasive cell numbers (P < 0.01). DDX11-AS1 knockdown inhibited SW780 cells growth in vivo and suppressed CDK6 and Ki67 expression in xenograft tumors. CONCLUSION: DDX11-AS1 exacerbates BLCA progression by enhancing CDK6 expression via suppressing miR-499b-5p.
PURPOSE: We investigated DDX11-AS1 effects on bladder cancer (BLCA) progression to identify a new potential therapeutic target for BLCA. METHODS: BLCA cases (n = 108) were enrolled. SW780 and J82 cells were transfected. Cell counting kit-8 (CCK-8) assay, wound healing assay and transwell migration assay was conducted. Cell cycle and apoptosis was detected by flow cytometry. Luciferase reporter assay was performed. DDX11-AS1, miR-499b-5p and CDK6 mRNA expression in tissues/cells was determined by quantitative real-time polymerase chain reaction (qRT-PCR). In vivo experiment was performed using nude mice. CDK6 and Ki67 proteins expression in cells and xenograft tumors were researched by Western blot and immunohistochemistry. RESULTS: Overexpressed DDX11-AS1 in BLCA was associated with poor outcome of patients. Compared with siCtrl group, SW780 and J82 cells of siDDX11-AS1 group had lower OD450 value (P < 0.01), less cells in S phase, more apoptosis cells (P < 0.05), higher relative wound width (P < 0.05) and less invasive cell number (P < 0.01). DDX11-AS1 promoted CDK6 expression via inhibiting miR-499b-5p. Compared with oe-DDX11-AS1 group, SW780 cells of oe-DDX11-AS1 + miR-499b-5p mimic group and oe-DDX11-AS1 + siCDK6 group had lower OD450 value (P < 0.01), less cells in S phrase, more apoptosis cells (P < 0.01), higher relative wound width (P < 0.05) and less invasive cell numbers (P < 0.01). DDX11-AS1 knockdown inhibited SW780 cells growth in vivo and suppressed CDK6 and Ki67 expression in xenograft tumors. CONCLUSION:DDX11-AS1 exacerbates BLCA progression by enhancing CDK6 expression via suppressing miR-499b-5p.