摘要
目的探讨IL-37对骨肉瘤细胞增殖、凋亡、迁移和侵袭的影响及其作用机制。方法定量逆转录PCR(RT-qPCR)检测人正常成骨细胞(hFOB1.19)和骨肉瘤细胞(U2OS、MG63、Saos-2)中IL-37 mRNA的表达情况;设计沉默IL-37的小干扰RNA(siRNA)序列转染骨肉瘤细胞系,将转染干扰序列siIL-37沉默IL-37表达的细胞作为siIL-37组,转染阴性对照序列作为阴性对照组(siNC组),检测沉默IL-37后骨肉瘤细胞IL-37 mRNA的表达。采用CCK-8法、Transwell法检测沉默IL-37表达对骨肉瘤细胞增殖、侵袭和迁移的影响;流式细胞术检测沉默IL-37表达对骨肉瘤细胞凋亡的影响;蛋白免疫印迹法检测沉默IL-37表达对骨肉瘤细胞中B淋巴细胞瘤(BCL)关联X蛋白(BAX)/BCL-2和Wnt/β-catenin信号通路中关键蛋白表达的影响。结果IL-37 mRNA在U2OS、MG63和Saos-2细胞中均高于其在hFOB1.19细胞中的表达(P均<0.05)。siIL-37组中的IL-37 mRNA相对表达量均低于siNC组(P均<0.05)。与siNC组比较,siIL-37组的细胞增殖比例下降,细胞凋亡率升高,迁移和侵袭细胞数量增多,BAX表达上调而BCL-2表达下调,Wnt/β-catenin信号通路中的关键蛋白c-Myc、β-catenin、转录增强因子1、细胞周期蛋白D1和基质金属蛋白酶-7表达均下调(P均<0.05)。结论IL-37在骨肉瘤细胞中表达升高,沉默IL-37表达可促进骨肉瘤细胞凋亡,并抑制其增殖、迁移和侵袭。IL-37可能通过BAX/BCL-2和Wnt/β-catenin信号通路调控骨肉瘤的发生发展。
Objective To investigate the effect of IL-37 on the proliferation,apoptosis,migration and invasion of osteosarcoma cells and its mechanism.Methods The expression of IL-37 in human normal osteoblasts(hFOB1.19)and osteosarcoma cell lines(U2OS,MG63,Saos-2)was detected by quantitative reverse transcription polymerase chain reaction(RT-qPCR).The siRNA sequence silencing IL-37 was designed and transfected into the osteosarcoma cell lines.Cells transfected with siRNA sequence silencing IL-37 were assigned into the siIL-37 group,and those transfected with negative controls were allocated into the siNC group.The expression of IL-37 mRNA in osteosarcoma cells after IL-37 silencing was detected.The effect of IL-37 silencing on the proliferation,invasion and migration of osteosarcoma cells was evaluated by CCK-8 and Transwell chamber test.The effect of IL-37silencing on the apoptosis of osteosarcoma cell lines was detected by flow cytometry.The expression levels of key proteins in the BAX/BCL-2 and Wnt/β-catenin signaling pathway of osteosarcoma cell lines before and after silencing IL-37 were determined western blot.Results The expression levels of IL-37 mRNA in osteosarcoma cell lines U2OS,MG63 and Saos-2 were significantly higher than that in normal osteoblast cell line(all P<0.05).For osteosarcoma cell lines U2OS,MG63 and Saos-2,the relative expression levels of IL-37 mRNA in the siIL-37 group were significantly lower than those in the siNC group(all P<0.05).Compared with the siNC group,cell proliferation rate was declined,cell apoptosis rate was increased,the quantity of cell migration and invasion was elevated,BAX expression was up-regulated,BCL-2 expression was down-regulated,and the expression levels of c-Myc,β-catenin,TEF1,Cyclin D1 and MMP-7 in the Wnt/β-catenin signal pathway were down-regulated in the siIL-37 group(all P<0.05).Conclusions The expression level of IL-37 is up-regulated in osteosarcoma cell lines.Silencing IL-37 can promote the apoptosis of osteosarcoma cells and inhibit the proliferation,migration and invasion.IL-37 plays an important carcinogenic role in the occurrence and development of osteosarcoma possibly through the BAX/BCL-2 and Wnt/β-catenin signaling pathways.
作者
陈海堡
宋毅昌
彭磊
曾统
张迪
靳松
Chen Haibao;Song Yichang;Peng Lei;Zeng Tong;Zhang Di;Jin Song(Department of Spinal Surgery,the Eighth Affiliated Hospital of Sun Yat-sen University,Shenzhen 518033,China)
出处
《新医学》
CAS
2022年第6期416-422,共7页
Journal of New Medicine
基金
深圳市福田区卫生公益性科研项目重点项目(FTWS2020025)
深圳市医学重点学科建设经费(脊柱外科立项SZXK086)。
