摘要
目的探讨骨保护素(OPG)/细胞核因子-κB受体活化因子(RANK)/细胞核因子-κB受体活化因子配体(RANKL)信号通路介导的自噬及微小RNA(miRNA,miR)-217的靶向调控在骨巨细胞瘤(GCT)中作用。方法在建立肿瘤异种移植模型和体外培养GCT0404细胞的基础上,分为空白对照组,神经钙黏蛋白(N-cadherin)组(miR-217类似物组),转染miR-217组。评估各组骨巨细胞瘤的活性、增殖及迁移水平;检测miR-217、OPG、RANK、RANKL、自噬相关蛋白5(ATG5)、自噬相关蛋白7(ATG7)、Beclin-1和微管相关蛋白轻链3(LC3)在骨巨细胞瘤细胞中的mRNA和蛋白表达变化;多组间均数比较采用单因素方差分析(ANOVA),组间比较采用多重极差最小显著性差异(LSD)。结果(1)miR-217在肿瘤异种移植模型和体外培养GCT0404细胞的调控都显著抑制GCT细胞的增殖和肿瘤细胞的发生[对照组吸光度(A)值为0.620±0.019,inhibitor NC组为0.640±0.019,miR-217 inhibitor组为0.950±0.030,inhibitorNC组和对照组比较,差异无统计学意义(P>0.05),而miR-217 inhibitor组与inhibitor NC组比较,差异有统计学意义(F=128.600,P<0.01)]。(2)miR-217抑制GCT细胞的碱性磷酸酶(ALP)活性,并抑制OPG/RANKL/RANK信号通路相关因子及自噬相关蛋白表达(OCN 1.41±0.06,OPG 1.41±0.04,RANKL 0.54±0.01,ATG 50.65±0.12,ATG 70.67±0.03,Beclin-10.56±0.10,LC30.83±0.04,F=265.100,P<0.01)。结论miR-217可能通过抑制OPG/RANKL/RANK信号通路相关因子表达来抑制GCT自噬,从而抑制GCT的发生。
Objective To explore the role of autophagy and targeted regulation of microRNA(miRNA,miR)-217 mediated by osteoprotegrin-receptor activator of nuclear factor-κB ligand-receptor activator of nuclear factor-κB(OPG-RANKL-RANK)signal pathway in giant cell tumor of bone(GCT).Methods On the basis of establishing tumor xenotransplantation model and culturing GCT0404 cells in vitro,they were divided into blank control group,N-cadherin group(miR-217 analogue group)and miR-217 group.To evaluate the activity,proliferation and migration of giant cell tumor of bone,and to detect the expression of miR-217,OPG,RANK,RANKL,autophagy associated protein 5(ATG5),autophagy associated protein 7(ATG7),Beclin-1 and microtubule associated protein light chain 3(LC3)in bone giant cell tumor cells.Single factor analysis of variance(ANOVA)was used to compare the mean among groups.Multiple minimum significant difference(LSD)was used for comparison between groups.Results(1)The regulation of miR-217 in tumor xenotransplantation model and in vitro cultured GCT0404 cells significantly inhibited the proliferation of GCT cells and the occurrence of tumor cells[In control group,optical density(A)value was 0.620±0.019 in NC inhibitor group,0.640±0.019 in inhibitor group,0.950±0.030 in inhibitor group,and there was no significant difference between miR-217 inhibitor group and control group,but there was significant difference between miR-217 inhibitor group and inhibitor NC group,F=128.600,P<0.01].(2)MiR-217 inhibited the activity of alkaline phosphatase(ALP)and the expression of OPG/RANKL/RANK signal pathway-related factors and autophagy-related proteins in GCT cells(OCN 1.41±0.06,OPG 1.41±0.04,RANKL 0.54±0.01,ATG 50.65±0.12,ATG 70.67±0.03,Beclin-10.56±0.10,LC30.83±0.04,F=265.100,P<0.01).Conclusion MiR-217 may inhibit GCT autophagy inhibiting the expression of factors related to OPG/RANKL/RANK signal pathway,thus inhibiting the occurrence of GCT.
作者
赵振群
梁晨亮
王玉鑫
赵伟
武利栓
崔淑霞
白锐
Zhao Zhenqun;Liang Chenliang;Wang Yuxin;Zhao Wei;Wu Lisuan;Cui Shuxia;Bai Rui(Department of Pediatric Orthopaedics,the Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,China;Graduate School,Inner Mongolia Medical University,Hohhot 010030,China;Department of Bone Oncology,the Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,China;Department of Anesthesiology,the Second Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010030,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2020年第11期2050-2054,共5页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81660457)。
