Objective:To investigate the expression profile of calcyphosine (CAPS) in bladder cancer and its role in regulating tumor cell biological behavior.Methods:From July 2023 to January 2024, CAPS-overexpressing and CAPS-silenced UM-UC-3 bladder cancer cell lines were constructed. The experimental groups were as follows: overexpression negative control (OvNC) group, transfected with empty lentiviral vector; overexpression group (OvCAPS), transfected with CAPS overexpression vector (pcDNA3.1-CAPS); knockdown groups (shCAPS.1 and shCAPS.2), transfected with two different lentiviral vectors encoding CAPS-specific short hairpin RNAs (shRNAs); and knockdown negative control (shNC), transfected with non-targeting shRNA lentiviral vector. Transfection efficiency was observed using fluorescence microscopy. CAPS mRNA and protein expression levels were detected by RT-qPCR and Western blotting, respectively. Cell proliferation, migration, and invasion capabilities were evaluated using cell counting kit-8 (CCK-8) and Transwell assays. Comparisons between groups were analyzed using the t-test. Results:CAPS protein expression differed significantly among experimental groups. Western blotting showed that CAPS protein levels in the OvCAPS group (0.61±0.03) were markedly higher than those in the OvNC group (0.38±0.02, t=10.89, P<0.001). In contrast, expression levels in the shCAPS.1 (0.21±0.01) and shCAPS.2 (0.20±0.01) groups were significantly lower than in the shNC group (0.60±0.04, t=14.72 and 14.45, respectively, P<0.001). Functional experiments further demonstrated that CAPS markedly influenced the biological behavior of bladder cancer cells. CCK-8 assays showed that the proliferation rate of the OvCAPS group at 48 h was significantly higher than that of the OvNC group (0.63±0.08 vs. 0.44±0.06, t=4.02, P<0.01), whereas proliferation was significantly reduced in knockdown groups as compared with shNC (shCAPS.1: 0.49±0.03 vs. 0.67±0.08, t=4.28, P<0.05; shCAPS.2: 0.43±0.04 vs. 0.67±0.08, t=5.99, P<0.05). Transwell migration assays revealed that the number of migrating cells in the OvCAPS group was significantly greater than that in the OvNC group (747.3±14.2 vs. 284.7±14.0, t=40.14, P<0.05), whereas the number of migrating cells was significantly reduced in the shCAPS.1 (116.3±3.2 vs. 327.3±7.6, t=44.43, P<0.05) and shCAPS.2 (137.0±3.0 vs. 327.3±7.6, t=40.48, P<0.05) groups. Similarly, invasion assays showed the same trend: the number of invading cells was significantly greater in the OvCAPS group (881.3±12.1 vs. 370.3±16.5, t=43.31, P<0.05) and markedly less in shCAPS.1 (96.0±1.0 vs. 367.0±10.0, t=37.45, P<0.05) and shCAPS.2 (121.0±5.3 vs. 367.0±10.0, t=29.46, P<0.05) grougs than in shNC group. Conclusion:CAPS is highly expressed in bladder cancer cells and may play a critical role in tumor progression by promoting cell proliferation, migration, and invasion.
(8.0+极速模式)