circCRKL靶向miR-942-5p对骨肉瘤Saos-2细胞增殖、迁移和侵袭的影响  

Effects of circCRKL targeting miR-942-5p on the proliferation,migration and invasion of osteosarcoma Saos-2 cells

作　　者：熊涛[1] 李秀山[1] XIONG Tao;LI Xiushan(Department of Orthopaedics,the First People's Hospital of Jingzhou,Jingzhou 434007,Hubei,China)

机构地区：[1]荆州市第一人民医院骨科,湖北荆州434007

出　　处：《中华骨与关节外科杂志》2024年第3期261-266,共6页Chinese Journal of Bone and Joint Surgery

摘　　要：目的:探讨circCRKL靶向miR-942-5p对骨肉瘤Saos-2细胞增殖、迁移和侵袭的影响。方法:回顾性分析2018年1月至2022年12月接受新辅助化疗的41例骨肉瘤患者的临床资料。所有患者在接受此次治疗前均未接受任何抗肿瘤治疗,手术时取骨肉瘤组织和瘤旁组织。采用RT-qPCR评估骨肉瘤组织中circCRKL、miR-942-5p表达情况。分别将pcDNA、pcDNA-circCRKL、si-NC、si-circCRKL、anti-miR-NC、pcDNA-circCRKL+miR-NC、anti-miR-942-5p、pcDNA-circCRKL+miR-942-5p转染Saos-2细胞,运用集落形成、划痕愈合、CCK-8和Transwell实验评估Saos-2细胞克隆数、划痕愈合率、细胞活力以及侵袭数。circCRKL和miR-942-5p的相互作用通过双荧光素酶报告基因法确定。结果:骨肉瘤组织中circCRKL表达下调(P<0.05),miR-942-5p表达上调(P<0.05)。过表达circCRKL后细胞克隆数、划痕愈合率、光密度(OD)值、侵袭数、miR-942-5p相对表达水平显著降低(P均<0.05)。抑制circCRKL表达后miR-942-5p相对表达水平显著升高(P<0.05)。下调miR-942-5p表达后细胞克隆数、划痕愈合率、OD值、侵袭数显著降低(P均<0.05)。circCRKL与miR-942-5p直接结合。上调miR-942-5p表达显著减弱circCRKL过表达对细胞克隆数、划痕愈合率、OD值、侵袭数的影响(P均<0.05)。结论:circCRKL通过靶向下调miR-942-5p来抑制骨肉瘤Saos-2细胞增殖、迁移和侵袭。Objective:To investigate the effect of circCRKL targeting miR-942-5p on the proliferation,migration and invasion of osteosarcoma Saos-2 cells.Methods:Retrospective analysis of clinical data of 41 patients with osteosarcoma who received neoadjuvant chemotherapy from January 2018 to December 2022.All patients did not receive any anti-tumor treatment before receiving this treatment,and bone sarcoma tissue and adjacent tissue were taken during surgery.The expression of circCRKL and miR-942-5p in osteosarcoma tissues was evaluated by RT-qPCR.The Saos-2 cells were transfected with pcDNA,pcDNA-circCRKL,si-NC,si-circCRKL,anti-miR-NC,pcDNA-circCRKL+miR-NC,anti-miR-942-5p,or pcDNA-circCRKL+miR-942-5prespectively.The colony formation,scratch healing,CCK-8 and Transwell experiments were performed to evaluate the clone number,scratch healing rate,cell viability and invasion ability of Saos-2 cells.The interaction between circCRKL and miR-942-5p was determined by the dual luciferase reporter assay.Results:The expression of circCRKL was down-regulated(P<0.05)while the expression of miR-942-5p was up-regulated(P<0.05)in osteosarcoma tissue.Overexpression of circCRKL significantly reduced the cell clone number,scratch healing rate,optical density(OD)value,invasion number and miR-942-5p expression level(all P<0.05).Inhibition of circCRKL expression significantly increased the expression level of miR-942-5p(P<0.05).Down-regulation of the miR-942-5p expression significantly reduced the cell clone number,scratch healing rate,OD value and invasion number(all P<0.05).circCRKL directly bound with miR-942-5p.Up-regulation of the miR-942-5p expression significantly reduced the effect of circCRKL overexpression on cell clone number,scratch healing rate,OD value and invasion number(all P<0.05).Conclusions:circCRKL inhibits the proliferation,migration and invasion of osteosarcoma Saos-2 cells by targeting and down-regulating miR-942-5p.

关 键 词：骨肉瘤 circCRKL 增殖 迁移 侵袭 miR-942-5p 

分 类 号：R681.8[医药卫生—骨科学]