circFBXL5通过靶向miR-515-5p影响膀胱癌T24细胞的恶性生物学行为及其分子机制

目的：探讨circFBXL5通过靶向miR-515-5p影响膀胱癌T24细胞的增殖、迁移、侵袭及其分子机制。方法：收集2020年4月至2020年6月间在苏州市中西医结合医院手术切除的41例膀胱癌组织及其癌旁组织，采用qPCR法检测circFBXL5、miR-515-5p的表达；双荧光素酶报告实验验证circFBXL5与miR-515-5p之间的靶向关系，体外培养人膀胱癌T24细胞，实验分为si-NC组、si-circFBXL5组、anti-miR-NC⁺si-circFBXL5组和si-circFBXL5⁺anti-miR-515-5p组；MTT法、细胞克隆形成实验、FCM、Transwell实验和WB法分别检测转染后T24细胞的增殖、细胞克隆形成、迁移、侵袭和凋亡及BAX、Bcl-2蛋白水平。结果：膀胱癌组织中circFBXL5呈高表达，miR-515-5p呈低表达(均P<0.05);circFBXL5靶向且负向调控miR-515-5p的表达；敲减circFBXL5后T24细胞的增殖抑制率、凋亡率和BAX蛋白水平均显著增高(均P<...

The effects of circFBXL5 on the malignant biological behaviors of bladder cancer T24 cells by targeting miR-515-5p and its molecular mechanisms

Objective: To investigate the effects of circFBXL5 on the proliferation, migration, and invasion of bladder cancer T24 cells by targeting miR-515-5p and its possible molecular mechanisms. Methods: 41 bladder cancer tissues and their adjacent tissues were collected at Suzhou Hospital of Integrated Traditional Chinese and Western Medicine from April 2020 to June 2020. The expressions of circFBXL5 and miR-515-5p were detected by qPCR. Dual-luciferase reporter assay was used to verify the targeting relationship between circFBXL5 and miR-515-5p. Human bladder cancer cells T24 were cultured in vitro and divided into si-NC group, si-circFBXL5 group, anti-miR-NC+si-circFBXL5 group, and si-circFBXL5+anti-miR-515-5p group. MTT assay, plate colony formation assay, flow cytometry, Transwell assay and Western blotting were used to detect cell proliferation, clone formation, migration, invasion, apoptosis and BAX and Bcl-2 protein levels of T24 cells after transfection, respectively. Results: circFBXL5 was highly expressed and miR-515-5p was lowly expressed in bladder cancer tissues (all P<0.05). circFBXL5 could negatively regulate the expression of miR-515-5p. After the knockdown of circFBXL5, the cell proliferation inhibition rate, apoptosis rate, and protein level of BAX were significantly increased (all P<0.05); the number of cell clone formation, migration, and invasion cells were decreased (all P<0.05); the protein level of Bcl-2 was significantly decreased (P <0.05). Knockdown of circFBXL5 and miR-515-5p simultaneously could partially reverse the inhibiting effect of circFBXL5 knockdown on T24 cells. Conclusion: circFBXL5 promoted the proliferation, migration, and invasion of bladder cancer T24 cells by inhibiting miR-515-5p expression. circFBXL5 and miR-515-5p may serve as potential molecular targets for bladder cancer treatment.