Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma <italic style="font-style: italic">via</italic> regulating immune cell cytotoxicity

Jinling YUAN; Xinjian CHEN; Kai LIAO; Hongyan PENG; Liping LI; Shuju ZHANG

doi:10.16016/j.1000-5404.202006005

您当前的位置：

首页 >

文章列表页 >

Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma via regulating immune cell cytotoxicity

Basic Medicine | 更新时间：2026-01-09

- Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma via regulating immune cell cytotoxicity
- Journal of Third Military Medical University Vol. 42, Issue 18, Pages: 1787-1794(2020)
- 作者机构：
  
  1.410000 长沙，中南大学湘雅三医院检验科
  2.510062 广州，南部战区空军医院血管内科
  3.410000 长沙，长沙血液中心
  4.410007 长沙，湖南省儿童医院儿科医学研究所
- 作者简介：
- 基金信息：
  
  the National Natural Science Foundation for Young Scholars Young Scholars of China(81900055);the Natural Science Foundation of Hunan Provence(2020JJ5283);the Health Commission Foundation of Hunan Province(B2019021);the Foundation of Hunan Children's Hospital(2019ZSJ)
- DOI：10.16016/j.1000-5404.202006005
  CLC： R394.3; R730.23; R738.1
- Received：01 June 2020，
  
  Revised：2020-08-12，
  
  Published：30 September 2020
- 稿件说明：
移动端阅览
Jinling YUAN, Xinjian CHEN, Kai LIAO, et al. Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma via regulating immune cell cytotoxicity[J]. Journal of Third Military Medical University, 2020, 42(18): 1787-1794.
DOI：

Jinling YUAN, Xinjian CHEN, Kai LIAO, et al. Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma via regulating immune cell cytotoxicity[J]. Journal of Third Military Medical University, 2020, 42(18): 1787-1794. DOI： 10.16016/j.1000-5404.202006005.

摘要

目的

探讨核心节律分子在骨肉瘤预后中的作用及可能机制。

方法

在骨肉瘤细胞中过表达节律基因BMAL1，qRT-PCR检测BMAL1基因的表达；克隆形成实验和Transwell实验检测细胞的克隆形成和侵袭能力；骨肉瘤细胞与外周血单个核淋巴细胞(peripheral blood mononuclear cell，PBMCs)体外共培养后，钙黄素染色法检测PBMCs对骨肉瘤细胞的直接杀伤作用，流式细胞术检测PBMCs中CD8

T和NK细胞脱颗粒(CD107a蛋白表达)的影响。

结果

BMAL1 mRNA高表达的骨肉瘤患者的生存时间明显高于低表达组(

0.000 1)。BMAL1过表达对U2OS细胞的侵袭和克隆形成无显著影响，但相较于对照组而言，过表达BMAL1的U2OS在体外与外周血单核血细胞(PBMCs)共培养4 h，其细胞死亡明显增加[PBMCs:U2OS细胞为5:1时，对照组和BMAL1组分别为(73.33±0.67)%和(81.00±0.58)%，

0.01；PBMCs:U2OS细胞为10:1时，对照组和BMAL1组分别为(76.67±0.33)%和(82.67±1.86)%，

0.01

]

。过表达BMAL1的骨肉瘤细胞组与PBMCs共培养后，T细胞和NK细胞CD107a表达差异无统计学意义(E:T=5:1，CD3

CD56

T细胞

=0.46; CD3

CD56

NK细胞

=0.96；E:T=10:1

CD3

CD56

T细胞

=0.93; CD3

CD56

NK细胞

=0.21)。

结论

骨肉瘤细胞BMAL1表达水平与骨肉瘤患者预后呈正相关，其机制主要是通过增加免疫细胞对其免疫监视作用，而非通过影响骨肉瘤细胞的克隆形成能力和侵袭。

Abstract

Objective

To investigate the role and possible mechanism of the core rhythm molecule BMAL1 in the prognosis of osteosarcoma (OS).

Methods

After BMAL1 was overexpressed in U2OS cells by Lipofectamine3000

the expression of BMAL1 was detected with qRT-PCR

and the clone formation and invasion ability of the overexpressed cells were observed with clonogenic assay and Transwell assay. Then

the BMAL1 overexpressed U2OS cells or the cells with control vector transfection were cocultured with peripheral blood mononuclear cells (PBMCs)

the direct killing effects of PBMCs on U2OS cells was observed with Calcein AM staining

and the CD107a expression by CD8

T and NK cells was measured with flow

cytometry.

Results

The OS patients with higher BMAL1 mRNA level had significantly longer survival time when compared with those with lower expression (

0.001). BMAL1 overexpression had no obvious effects on the invasion and clone formation of U2OS cells. However

cell death was significantly increased in BMAL1 overexpressed U2OS cells than the control vector transfected U2OS cells when co-cultured with PBMCs

in vitro

[PBMCs:U2OS cells=5:1

(81.00±0.58)%

(73.33±0.67)%

0.01; PBMCs:U2OS cells=10:1

(82.67±1.86)%

(76.67±0.33)%

0.01

]

. There was no significant difference in the expression level of CD107a between T cells and NK cells in the BMAL1 overexpressed U2OS cells than the control vector transfected U2OS cells when co-cultured with PBMCs (E:T=5:1

CD3

CD56

T cells

=0.46

CD3

CD56

NK cells

=0.96; E:T=10:1

CD3

CD56

T cells

=0.93; CD3

CD56

NK cells

=0.21).

Conclusion

The expression level of BMAL1 is positively correlated with the prognosis of OS patients

which might be mainly due to the enhanced immunological surveillance

but not by affecting the ability of invasion and clone formation of OS cells.

关键词

Keywords

references

BIERMANN J S, ADKINS D R, AGULNIK M, et al .Bone cancer[J ] . J Natl Compr Canc Netw, 2013, 11(6): 688-723. DOI: 10.6004/jnccn.2013.0088.

NG A J, MUTSAERS A J, BAKER E K, et al . Genetically engineered mouse models and human osteosarcoma[J ] . Clin Sarcoma Res, 2012, 2(1): 1-14. DOI: 10.1186/2045-3329-2-19.

ALLISON D C, CARNEY S, AHLMANNE R, et al . A meta-analysis of osteosarcoma outcomes in the modern medical era[J ] . Sarcoma, 2012, 2012: 704872. DOI: 10.1155/2012/704872.

LONGHI A, ERRANI C, DE PAOLIS M, et al . Primary bone osteosarcoma in the pediatric age: state of the art[J ] . Cancer Treat Rev, 2006, 32(6): 423-436. DOI: 10.1016/j.ctrv.2006.05.005.

KAGER L, ZOUBEK A, POTSCHGER U, et al . Primary metastatic osteosarcoma: presentation and outcome of patients treated on neoadjuvant cooperative osteosarcoma study group protocols[J ] . J Clin Oncol, 2003, 21(10): 2011-2018. DOI: 10.1200/JCO.2003.08.132.

KAGER L, TAMAMYAN G, BIELACK S. Novel insights and therapeutic interventions for pediatric osteosarcoma[J]. Futur Oncol, 2017, 13(4): 357-368. DOI: 10.2217/fon- 2016-0261.

JONES K B. Osteosarcoma genesis: modeling cancer initiation in the mouse[J]. Sarcoma, 2011, 2011: 694136. DOI: 10.1155/2011/694136.

LUSSIER D M, JOHNSON J L, HINGORANI P, et al . Combination immunotherapy with α-CTLA-4 and α-PD-L1 antibody blockade prevents immune escape and leads to complete control of metastatic osteosarcoma[J ] . J Immunother Cancer, 2015, 3: 21. DOI: 10.1186/s40425-015-0067-z. eCollection 2015.

HEYMANN M F, SCHIAVONE K, HEYMANN D.Bone sarcomas in the immunotherapy era[J]. Br J Pharmacol, 2020, 23.DOI: 10.1111/bph.14999.

BASS J, LAZAR M A. Circadian time signatures of fitness and disease[J]. Science, 2016, 354(6315): 994-999. DOI: 10.1126/science.aah4965.

PANDA S. Circadian physiology of metabolism[J]. Science,2016, 354(6315): 1008-1015. DOI: 10.1126/science. aah4967.

FU L, PELICANO H, LIU J S, et al . The circadian gene Period2 plays an important role in tumor suppression and DNA damage response in vivo[J ] . Cell, 2002, 111(1): 41-50. DOI: 10.1016/s0092-8674(02)00961-3.

LOGAN R W, ZHANG C Q, MURUGAN S, et al . Chronic shift-lag alters the circadian clock of NK cells and promotes lung cancer growth in rats[J ] . J Immunol, 2012, 188(6): 2583-2591. DOI: 10.4049/jimmunol.1102715.

WU Y C, TAO B R, ZHANG T Y, et al . Pan-cancer analysis reveals disrupted circadian clock associates with T cell exhaustion[J ] . Front Immunol, 2019, 10: 2451. DOI: 10.3389/fimmu.2019.02451.

KIM E, JUNG S, PARK W S, et al .Upregulation of SLC2A3 gene and prognosis in colorectal carcinoma: analysis of TCGA data[J ] . BMC Cancer, 2019, 19(1): 302. DOI: 10.1186/s12885-019-5475-x.

ZHANG S J, ZHANG J M, DENG Z Y, et al .Circadian clock components RORα and BMAL1 mediate the anti-proliferative effect of MLN4924 in osteosarcoma cells[J ] . Oncotarget, 2016, 7(40): 66087-66099. DOI: 10.18632/oncotarget.11807.

GATTI R, BELLETTI S, ORLANDINI G, et al . Comparison of annexin V and calcein-AM as early vital markers of apoptosis in adherent cells by confocal laser microscopy[J ] . J Histochem Cytochem, 1998, 46(8): 895-900. DOI: 10.1177/002215549804600804.

AKTAS E, KUCUKSEZER U C, BILGIC S, et al .Relationship between CD107a expression and cytotoxic activity[J ] . Cell Immunol, 2009;254(2): 149-154. DOI: 10.10 16/j.cellimm. 2008.08.007.

GERY S, KOMATSU N, BALDJYAN L, et al .The circadian gene per1 plays an important role in cell growth and DNA damage control in human cancer cells[J ] . Mol Cell, 2006, 22(3): 375-382. DOI: 10.1016/j.molcel.2006.03.038.

ZENG Z L, LUO H Y, YANG J, et al . Overexpression of the circadian clock gene BMAL1 increases sensitivity to oxaliplatin in colorectal cancer[J ] . Clin Cancer Res, 2014, 20(4): 1042-1052. DOI: 10.1158/1078-0432.CCR-13-0171.

ZHANG F, SUN H, ZHANG S, et al . Overexpression of PER3 inhibits self-renewal capability and chemoresistance of colorectal cancer stem-like cells via inhibition of notch and β-catenin signaling[J ] . Oncol Res, 2017, 25(5): 709-719. DOI: 10.3727/096504016X14772331883976.

PURAM R V, KOWALCZYK M S, DE BOER C G, et al . Core circadian clock genes regulate leukemia stem cells in AML[J ] . Cell, 2016, 165(2): 303-316. DOI: 10.1016/j.cell.2016.03.015.

XIA W K, LIU Z L, SHEN D, et al . Prognostic performance of pre-treatment NLR and PLR in patients suffering from osteosarcoma[J ] . World J Su rg Oncol, 2016, 14(1): 1-8. DOI: 10.1186/s12957-016-0889-2.

PAHL J H, KWAPPENBERG K M, VARYPATAKI E M, et al .Macrophages inhibit human osteosarcoma cell growth after activation with the bacterial cell wall derivative liposomal muramyl tripeptide in combination with interferon-γ[J ] . J Exp Clin Cancer Res, 2014, 33(1): 27. DOI: 10.1186/1756-9966-33-27.

LABRECQUE N, CERMAKIAN N.Circadian clocks in the immune system[J]. J Biol Rhythms, 2015, 30(4): 277-290. DOI: 10.1177/0748730415577723.

NOBIS C C, DUBEAU LARAMÉE G, KERVEZEE L, et al . The circadian clock of CD8 T cells modulates their early response to vaccination and the rhythmicity of related signaling pathways[J ] . Proc Natl Acad Sci USA, 2019, 116(40): 20077-20086. DOI: 10.1073/pnas.1905080116.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Expression of miR-616 in osteosarcoma and its role in proliferation, apoptosis, migration and invasion of tumor cells

Application of 3D-printed customized prostheses in limb-salvage surgery for pediatric osteosarcoma of the distal femur: a case series study

Angiotensin Ⅱ type 1 receptor autoantibody-AT1R-Bmal1 axis promotes phenotypic transition of vascular smooth muscle cells and vascular fibrosis

Sonodynamic cytotoxicity of sparfloxacin on rheumatoid arthritis fibroblast-like synoviocytes in vitro

Effect of overexpression of imprinted gene TSSC3 on angiogenesis in human osteosarcoma

Related Author

FU Wanlei

HAO Xianglin

CAO Ya

XIA Jiying

ZHOU Xiaogang

XU Jiayi

GUO Qiaonan

QIAN Fusheng

Related Institution

Department of Pathology， Second Affiliated Hospital， Army Medical University （Third Military Medical University）

Department of Orthopedics， Affiliated Children’s Hospital， Chongqing Medical University

School of Basic Medical Sciences， Shanxi Medical University

Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Chongqing Medical University

Department of Pharmacy, the First Affiliated Hospital of Chongqing Medical University

⁰

Circadian rhythms molecule BMAL1 promotes the prognosis of osteosarcoma via regulating immune cell cytotoxicity

DOI：10.16016/j.1000-5404.202006005

摘要

Abstract

关键词

Keywords

references