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

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

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<0.05),细胞克隆形成数和迁移、侵袭细胞数均显著减少(均P<0.05),Bcl-2蛋白水平显著降低(P<0.05);同时敲减circFBXL5和miR-515-5p可部分逆转敲减circFBXL5对T24细胞增殖的抑制作用。结论:circFBXL5通过调控miR-515-5p表达影响膀胱癌T24细胞的增殖、迁移、侵袭,circFBXL5和miR-515-5p可能膀胱癌治疗的潜在分子靶标。

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,sicircFBXL5 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.