[1]罗　凯,段华彬,罗明鼎,等.HOXB3促进骨肉瘤细胞增殖、克隆形成、迁移和抑制细胞凋亡的机制研究[J].现代检验医学杂志,2022,37(01):136-140.[doi:10.3969/j.issn.1671-7414.2022.01.027]
　LUO Kai,DUAN Hua-bin,LUO Ming-ding,et al.Mechanism of HOXB3 Promoting Proliferation, Cloning Formation, Migration and Inhibiting Apoptosis of Osteosarcoma Cells[J].Journal of Modern Laboratory Medicine,2022,37(01):136-140.[doi:10.3969/j.issn.1671-7414.2022.01.027]
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HOXB3促进骨肉瘤细胞增殖、克隆形成、迁移和抑制细胞凋亡的机制研究()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第37卷
期数:
2022年01期
页码:
136-140
栏目:
论　著
出版日期:
2022-01-15

文章信息/Info

Title:
Mechanism of HOXB3 Promoting Proliferation, Cloning Formation, Migration and Inhibiting Apoptosis of Osteosarcoma Cells
文章编号:
1671-7414（2022）01-136-06
作者:
罗　凯; 段华彬; 罗明鼎; 宋世杰
（雅安市雨城区人民医院骨科，四川雅安 625000）
Author(s):
LUO Kai;  DUAN Hua-bin;  LUO Ming-ding;  SONG Shi-jie
（Department of Orthopedics, the Yucheng Disrict People’s Hospital of Ya’an, Sichuan Ya’an 625000, China）
关键词:
骨肉瘤; 同源异型盒基因 B3; 染色质免疫共沉淀技术 3; 增殖; 克隆形成; 迁移; 凋亡
分类号:
R738.1；R730.43
DOI:
10.3969/j.issn.1671-7414.2022.01.027
文献标志码:
A
摘要:
目的　探究同源异型盒基因 B3（homeobox genes B3, HOXB3）对骨肉瘤细胞增殖、克隆形成、迁移和凋亡的影响及其潜在分子机制。方法　检测 2020年 1月～ 12月在雅安市第二人民医院骨科行手术治疗的 30例骨肉瘤患者病理组织及邻近正常组织中 HOXB3表达；通过转染干扰 siRNA敲低 HOXB3表达，验证其转染效率；采用细胞增殖实验、克隆形成实验、细胞划痕实验及细胞凋亡实验分别探究敲低 HOXB3对骨肉瘤细胞增殖、克隆形成、迁移和凋亡的影响；分析 CDCA3在骨肉瘤中的表达特征及与 HOXB3的相关性，通过染色质免疫共沉淀技术（ chromatin immunoprecipitation, ChIP）分析和荧光素酶报告基因实验验证 HOXB3和细胞分裂周期相关蛋白 3（cell division cycle associatedfprotein 3, CDCA3）结合关系；验证 HOXB3和 CDCA3表达调控对骨肉瘤细胞生物学行为的作用机制。结果　30例骨肉瘤临床组织中 HOXB3表达（ 41.154±8.626）较邻近正常组织（22.857±7.512）显著升高，差异有统计学意义（t=8.975，P＜ 0.001）。敲低 HOXB3表达后，骨肉瘤细胞增殖率、克隆形成率和迁移率明显降低（ F=37.477～ 399.842，均 P＜ 0.001），细胞凋亡率显著升高（F=10.556，P=0.011）。骨肉瘤组织中CDCA3表达（ 38.624±7.736）明显高于邻近正常组织（21.875±6.482），差异有统计学意义（ t=9.090，P＜ 0.001）；HOXB3与 CDCA3表达呈正相关（ r=0.736，P＜ 0.01）。CDCA3是 HOXB3的靶基因；敲低 HOXB3表达显著降低了骨肉瘤细胞中 CDCA3表达（ F=694.283，P＜ 0.001）。HOXB3可结合 CDCA3的启动子区域正调控 CDCA3的表达；在敲低 HOXB3表达的细胞中回补 CDCA3，逆转了 HOXB3对骨肉瘤细胞增殖、克隆形成、迁移、凋亡的抑制 /促进作用。结论　HOXB3在骨肉瘤中表达上调，其可能通过与 CDCA3启动子区域结合，正向调控 CDCA3表达促进了骨肉瘤细胞的增殖、克隆形成及迁移，抑制了细胞凋亡，可作为临床骨肉瘤治疗的潜在药物靶点。
Abstract:
Objective　To investigate the effects of HOXB3 on proliferation, clone formation, migration and apoptosis ofosteosarcoma cells and its potential molecular mechanism. Methods　HOXB3 expression in pathological tissues and adjacentnormal tissues of 30 patients with osteosarcoma who underwent surgical treatment in the Department of Orthopedics of theSecond People’s Hospital of Ya’an from January 2020 to December 2020 was detected. The expression of HOXB3 was knockeddown by interfering siRNA transfection, and the transfection efficiency was verified. The effects of knockdown HOXB3 onproliferation, clone formation, migration and apoptosis of osteosarcoma cells were investigated by cell proliferation assay, cloneformation assay, cell scratch assay and apoptosis assay.The expression characteristics of CDCA3 in osteosarcoma and itscorrelation with HOXB3 were analyzed.The binding relationship between HOXB3 and CDCA3 was verified by Chchip analysisand luciferase reporter assay. To verify the mechanism of HOXB3 and CDCA3 expression regulation on the biological behaviorof osteosarcoma cells. Results　HOXB3 expression in 30 cases of osteosarcoma (41.154±8.626) was significantly higher thanthat in adjacent normal tissues (22.857±7.512)，the difference was statistically significant (t=8.975, P＜ 0.001). Afterknockdown of HOXB3 expression, the proliferation rate, clone formation rate and migration rate of osteosarcoma cells weresignificantly decreased (F=37.477~399.842, P ＜ 0.001), and the apoptosis rate was significantly increased (F=10.556, P=0.011).CDCA3 expression in osteosarcoma tissues (38.624±7.736) was significantly higher than that in adjacent normal tissues(21.875±6.482), the difference was statistically significant (t=9.090, P ＜ 0.001). HOXB3 was positively correlatedwith CDCA3 expression (r=0.736, P ＜ 0.01). CDCA3 was the target gene of HOXB3. Knocking down HOXB3 expressionsignificantly decreased CDCA3 expression in osteosarcoma cells (F=694.283, P ＜ 0.001). HOXB3 could bind to the promoterregion of CDCA3 to positively regulate the expression of CDCA3. CDCA3 supplementation in the cells that knocked downHOXB3 expression reversed the inhibition/promotion of HOXB3 on proliferation, clone formation, migration and apoptosis ofosteosarcoma cells.Conclusion　HOXB3 expression is up-regulated in osteosarcoma. HOXB3 may positively regulate theexpression of CDCA3 by binding to the CDCA3 promoter region to promote the proliferation, clonal formation and migration ofosteosarcoma cells, and inhibit cell apoptosis. HOXB3 can be used as a potential drug target for clinical treatment ofosteosarcoma.

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备注/Memo

备注/Memo:
作者简介：罗凯（1980-），男，硕士，副主任医师，骨科，研究方向：脊柱退行性疾病，E-mail:hh1171989yjr@163.com。
更新日期/Last Update: 1900-01-01