摘要
目的探讨丙泊酚对膀胱癌细胞迁移及侵袭的影响以及其与微小RNA-383-5p(miR-383-5p)/蛋白磷酸酶2A的癌性抑制因子(CIP2A)轴的关系。方法体外培养膀胱癌细胞,分别用0、5、10和20μg/mL丙泊酚进行处理,筛选出丙泊酚的最佳浓度,将膀胱癌细胞分为对照组、丙泊酚组、丙泊酚+miR-NC组、丙泊酚+miR-383-5p组、丙泊酚+si-con组、丙泊酚+si-CIP2A组、丙泊酚+miR-383-5p+pcDNA组和丙泊酚+miR-383-5p+pc-CIP2A组。CCK-8法和克隆形成实验检测膀胱癌细胞增殖能力;Transwell实验评估膀胱癌细胞的侵袭、迁移能力;实时荧光定量聚合酶链式反应(qRT-PCR)测定miR-383-5p表达;蛋白免疫印迹法评价CIP2A蛋白表达;双荧光素酶报告基因实验证实miR-383-5p与CIP2A的靶向关系。结果20μg/mL丙泊酚组膀胱癌细胞增殖、侵袭和迁移能力均低于0、5、10μg/mL组,因此选择20μg/mL丙泊酚进行下一步实验。丙泊酚组膀胱癌细胞存活率、克隆形成率、侵袭和迁移细胞数及miR-383-5p表达均较对照组降低,CIP2A蛋白表达较对照组升高(P<0.05)。与丙泊酚组比较,丙泊酚+miR-383-5p组、丙泊酚+si-CIP2A组细胞存活率、克隆形成率、迁移和侵袭细胞数及miR-383-5p表达升高,CIP2A蛋白表达降低(P<0.05)。与丙泊酚+miR-383-5p组比较,丙泊酚+miR-383-5p+pc-CIP2A组细胞存活率、克隆形成率、侵袭和迁移细胞数下降,CIP2A蛋白表达升高(P<0.05)。结论丙泊酚被证明能够抑制膀胱癌细胞侵袭、迁移能力,并初步阐释了其抗癌作用可能与下调miR-383-5p、靶向提高CIP2A表达有关。
Objective To investigate the effects of propofol on the migration and invasion of bladder cancer cells and its relationship with the microRNA-383-5p(miR-383-5p)/cancerous inhibitor of protein phosphatase 2A(CIP2A)axis.Methods Bladder cancer cells were cultured in vitro and treated with 0,5,10,and 20μg/mL propofol,respectively,to screen the optimal concentration of propofol,and the bladder cancer cells were divided into control group,propofol group,and propofol+miR-NC group,propofol+miR-383-5p group,propofol+si-con group,propofol+si-CIP2A group,propofol+miR-383-5p+pcDNA group and propofol+miR-383-5p+pc-CIP2A group.The CCK-8 assay and clonogenesis assay were used to detect the proliferation of bladder cancer cells;the invasion and migration abilities of bladder cancer cells were evaluated by Transwell assay;the expression level of miR-383-5p was determined by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR);the expression level of CIP2A protein was evaluated by western blot;dual luciferase reporter gene assay confirmed the targeting relationship between miR-383-5p and CIP2A.Results The proliferation,invasion and migration of bladder cancer cells in the 20μg/mL propofol group were lower than those in the 0,5,and 10μg/mL groups,therefore,20μg/mL propofol was selected for the next experiment.Compared with the control group,the survival rate,clone formation rate,numbers of migrating and invasive cells,and the miR-383-5p expression of bladder cancer cells in the propofol group were reduced,the expression level of CIP2A protein was increased(P<0.05).compared with the propofol group,the survival rate,clone formation rate,numbers of migrating and invasive cells,and the miR-383-5p expression in the propofol+miR-383-5p group and the propofol+si-CIP2A group were increased,the CIP2A protein expression was reduced(P<0.05).Compared with propofol+miR-383-5p group,the survival rate,clone formation rate,and the numbers of migrating and invasive cells in the propofol+miR-383-5p+pc-CIP2A group were reduced,the CIP2A protein expression was increased(P<0.05).Conclusion Propofol was proved to inhibit the invasion and migration of bladder cancer cells,and it was preliminarily explained that its anticancer effect may be related to down-regulation of miR-383-5p and targeted enhancement of CIP2A expression.
作者
王刚
朱洪宽
陈永伦
李建钢
WANG Gang;ZHU Hong-kuan;CHEN Yong-lun(Department of Anesthesiology,Qujing First People’s Hospital,Qujing 655000,China)
出处
《中国实验诊断学》
2023年第3期341-345,共5页
Chinese Journal of Laboratory Diagnosis
