多发性骨髓瘤细胞通过PI3K/AKT信号通路促进M2巨噬细胞极化的机制研究

doi:10.12114/j.issn.1007-9572.2023.0322

中国全科医学 ›› 2024, Vol. 27 ›› Issue (08): 978-984.DOI: 10.12114/j.issn.1007-9572.2023.0322

• 论著·专病管理·多发性骨髓瘤 • 上一篇下一篇

多发性骨髓瘤细胞通过PI3K/AKT信号通路促进M2巨噬细胞极化的机制研究

彭逸伦¹^,², 李杨¹^,², 王晓桃¹^,²^,³^,^*()

1．541001　广西壮族自治区桂林市，桂林医学院临床医学院
2．541001　广西壮族自治区桂林市，桂林医学院附属医院临床试验中心
3．541001　广西壮族自治区桂林市，桂林医学院附属医院血液内科

收稿日期:2023-05-29 修回日期:2023-07-03 出版日期:2024-03-15 发布日期:2023-12-19
通讯作者: 王晓桃
作者贡献：王晓桃提出研究思路，设计研究方案，负责论文起草、最终版本的修订，对论文负责；彭逸伦进行研究过程的实施，包括进行细胞实验、流式细胞术实验，荧光定量PCR实验，调查对象的选取、样本的采集等；李杨负责数据收集、采集、清洗和统计学分析等。
基金资助:
国家自然科学基金资助项目(82060044); 广西自然科学基金资助项目(2020GXNSFAA159018); 国家级大学生创新创业训练计划(202010601010)

Study on the Mechanisms of Multiple Myeloma Cells Promoting M2 Macrophage Polarization through PI3K/AKT Signaling Pathway

PENG Yilun¹^,², LI Yang¹^,², WANG Xiaotao¹^,²^,³^,^*()

1. Clinical Medical School, Guilin Medical University, Guilin 541001, China
2. Clinical Trials Center, Affiliated Hospital of Guilin Medical University, Guilin 541001, China
3. Department of Hematopathology, Affiliated Hospital of Guilin Medical University, Guilin 541001, China

Received:2023-05-29 Revised:2023-07-03 Published:2024-03-15 Online:2023-12-19
Contact: WANG Xiaotao

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摘要/Abstract

摘要： 背景多发性骨髓瘤发病率长期居高不下，但目前关于磷脂酰肌醇-3-激酶（PI3K）/丝氨酸/苏氨酸激酶（AKT）信号通路和M2巨噬细胞极化促进多发性骨髓瘤发生进展的研究较少。目的探讨M2巨噬细胞在多发性骨髓瘤患者中的表达及PI3K/AKT信号通路促进M2巨噬细胞极化的机制研究。方法选取2021年10月—2022年4月于桂林医学院附属医院血液内科确诊的多发性骨髓瘤患者48例为试验组，选取同期健康志愿者（血液内科骨髓移植健康供者）30名为对照组。取2组研究对象外周血并分离单个核细胞，流式细胞术检测M2巨噬细胞比例。采用细胞传代培养RPMI8226细胞，通过佛波酯分化培养单核巨噬细胞THP-1为巨噬细胞。根据实验需要将瘤细胞培养并采用siRNA转染沉默磷酸酶基因（PTEN）分成3组：空白组、siRNA-PTEN实验组、siRNA对照组；收集以上3组细胞培养基上清液加入巨噬细胞共培养体系后分为4组：M0巨噬细胞组、瘤细胞上清液组、siRNA-PTEN上清液组、siRNA上清液组。采用Western blot实验检测多发性骨髓瘤细胞培养的空白组、siRNA-PTEN实验组、siRNA对照组的AKT、p-AKT、PI3K-p85、p-PI3K-p85蛋白表达水平。采用荧光定量聚合酶链式反应（PCR）检测多发性骨髓瘤细胞培养的空白组、siRNA-PTEN实验组、siRNA对照组的基质金属蛋白酶（MMP）2、MMP9的mRNA表达水平。采用流式细胞术检测多发性骨髓瘤上清液共培养的M0巨噬细胞组、瘤细胞上清液组、siRNA-PTEN上清液组、siRNA上清液组M2巨噬细胞中特异性抗体CD163、CD206、F4/80表达水平。采用荧光定量PCR检测多发性骨髓瘤上清液共培养的M0巨噬细胞组、瘤细胞上清液组、siRNA-PTEN上清液组、siRNA上清液组M2巨噬细胞特异标志物精氨酸酶1（ARG-1）、白介素10（IL-10）的mRNA表达水平。结果试验组M2巨噬细胞表达水平高于对照组（t=0.855，P<0.001）。siRNA-PTEN实验组中p-AKT/AKT、p-PI3K-p85/PI3K-p85蛋白表达水平均高于空白组和siRNA对照组（P<0.05）。siRNA-PTEN实验组中MMP2、MMP9的mRNA表达水平均高于空白组和siRNA对照组（P<0.05）。siRNA-PTEN上清液组M2巨噬细胞特异性抗体CD163+F4/80、CD206+F4/80表达水平高于瘤细胞上清液组和siRNA上清液组（P<0.05）。siRNA-PTEN上清液组M2巨噬细胞特异标志物ARG-1、IL-10的mRNA表达水平高于瘤细胞上清液组和siRNA上清液组（P<0.05）。结论多发性骨髓瘤患者中M2巨噬细胞表达上调，多发性骨髓瘤细胞可通过激活PI3K/AKT信号通路促进M2巨噬细胞极化，调控骨髓瘤细胞微环境。

关键词: 多发性骨髓瘤, 巨噬细胞, 信号传导, 肿瘤微环境, 生物标记，肿瘤, 细胞极化, 发病机制

Abstract:

Background

The incidence of multiple myeloma has long been high, however, there are fewer studies on phosphatidylinositol 3-kinase (PI3K) /serine-threonine kinase (AKT) signaling pathway and M2 macrophage polarization promoting the progression of multiple myeloma.

Objective

To investigate the expression of M2 macrophages in patients with multiple myeloma and the mechanism of PI3K/AKT signaling pathway promoting M2 macrophage polarization.

Methods

Forty-eight multiple myeloma patients diagnosed in the Department of Hematology of the Affiliated Hospital of Guilin Medical University from October 2021 to April 2022 were selected as the test group, and thirty healthy volunteers (healthy donors of blood marrow transplantation) in the same period were selected as the control group. Peripheral blood was taken from the 2 group and mononuclear cells were isolated, the proportion of M2 macrophages was determined by flow cytometry. RPMI8226 cells were subcultured by cell passage, and mononuclear macrophages THP-1 cells were cultured as macrophages by phorbol ester differentiation. According to the experimental requirements, tumor cells were cultured and transfected with siRNA to silence phosphatase and tensin homolog (PTEN) and divided into three groups, including blank group, siRNA-PTEN experimental group, and siRNA control group; the supernatants of the above three groups were collected and added to the macrophage co-culture system and then divided into four groups including M0 macrophage group, tumor cell supernatant group, siRNA-PTEN supernatant group, and siRNA supernatant group. The protein expression levels of AKT, p-AKT, PI3K-p85, and p-PI3K-p85 in the blank group, siRNA-PTEN experimental group, and siRNA control group of multiple myeloma cell culture were determined by Western blot assay. The mRNA expression levels of MMP2 and MMP9 in the blank group, siRNA-PTEN experimental group and siRNA control group were determined by fluorescence quantitative polymerase chain reaction (PCR) . The expression levels of specific antibodies CD163, CD206, and F4/80 in M2 macrophages in M0 macrophage group, tumor cell supernatant group, siRNA-PTEN supernatant group, and siRNA supernatant group were determined by flow cytometry. The mRNA expression levels of arginase 1 (ARG-1) and interleukin 10 (IL-10) in M0 macrophage group, tumor cell supernatant group, siRNA-PTEN supernatant group, and siRNA supernatant group were determined by fluorescence quantitative PCR.

Results

The expression level of M2 macrophages was higher in the test group than that in the control group (t=0.855, P<0.001) . The protein expression levels of p-AKT/AKT and p-PI3K-p85/PI3K-p85 in the siRNA-PTEN experimental group were higher than the blank group and siRNA control group (P<0.05) . The mRNA expression levels of MMP2 and MMP9 in the siRNA-PTEN experimental group were higher than the blank group and siRNA control group (P<0.05) . M2 macrophage specific antibody CD163+F4/80 and CD206+F4/80 in the siRNA-PTEN supernatant group were higher than the tumor cell supernatant and siRNA-PTEN supernatant group (P<0.05) . The mRNA expression levels of ARG-1 and IL-10 in the siRNA-PTEN supernatant group were higher than the tumor cell supernatant group and siRNA supernatant group (P<0.05) .

Conclusion

M2 macrophages are upregulated in multiple myeloma patients, and multiple myeloma cells can promote M2 macrophage polarization through activation of the PI3K/AKT signaling pathway, thereby regulating the myeloma cell microenvironment.

Key words: Multiple myeloma, Macrophages, Signal transduction, Tumor microenvironment, Biomarkers, tumor, Cell polarization, Pathogenesis

彭逸伦,李杨,王晓桃. 多发性骨髓瘤细胞通过PI3K/AKT信号通路促进M2巨噬细胞极化的机制研究[J]. 中国全科医学, 2024, 27(08): 978-984. DOI: 10.12114/j.issn.1007-9572.2023.0322.
PENG Yilun,LI Yang,WANG Xiaotao. Study on the Mechanisms of Multiple Myeloma Cells Promoting M2 Macrophage Polarization through PI3K/AKT Signaling Pathway[J]. Chinese General Practice, 2024, 27(08): 978-984. DOI: 10.12114/j.issn.1007-9572.2023.0322.

图/表 9

表1 实时定量PCR使用引物列表

Table 1 List of primers used for real-time quantitative PCR

基因名称	NCBIGeenID	基因编号	引物序列（5´-3´）	产物长度（bp）
ARG-1	383	NM_000045	TGGACAGACtAGGAATTGGCA	102
			CCAGTCCGTCAACATCAAAACt
IL-10	3586	NM_000572	GACTTTAAGGGTTACCTGGGTTG	112
			TCACATGCGCCTTGATGTCTG
MMP2	4313	NM_004530	GATACCCCTTTGACGGTAAGGA	112
			CCTTCTCCCAAGGTCCATAGC
MMP9	4318	NM_004994	GGGACGCAGACATCGTCATC	139
			TCGTCATCGTCGAAATGGGC
PTEN	5728	NM_000314	TTTGAAGACCATAACCCACCAC	134
β-actin	60	NM_001101	CATGTACGTTGCTATCCAGGC	250
			CTCCTTAATGTCACGCACGAT

图1 研究对象外周血M2巨噬细胞表达水平注：A为对照组外周血M2巨噬细胞（CD163和CD206的巨噬细胞）表达水平的流式细胞图；B为试验组外周血M2巨噬细胞（CD163和CD206的巨噬细胞）表达水平的流式细胞图。

Figure 1 Expression levels of M2 macrophages in peripheral blood

图2 倒置显微镜下RPMI8226细胞经siRNA转染后的细胞（荧光染色，×40）

Figure 2 The cellular level of RPMI8226 cells transfected with siRNA under inverted microscope

图3 多发性骨髓瘤细胞PI3K/AKT蛋白表达图注：AKT=丝氨酸/苏氨酸激酶，PI3K=磷脂酰肌醇-3-激酶。

Figure 3 Protein expression map of PI3K/AKTin multiple myeloma cells

表2 多发性骨髓瘤细胞中PI3K/AKT蛋白表达水平比较（±s，n=3）

Table 2 Comparison of protein expression level of PI3K/AKT in multiple myeloma cells

组别	p-AKT/AKT	p-PI3K-p85/PI3K-p85
空白组	1.070±0.070	0.843±0.287
siRNA-PTEN实验组	2.051±0.297^a	1.801±0.111^a
siRNA对照组	1.229±0.344^b	0.950±0.079^b
F值	11.795	24.579
P值	0.008	P<0.001

表3 多发性骨髓瘤细胞MMP2、MMP9的mRNA表达水平比较（±s，n=3）

Table 3 Comparison of the mRNA expression levels of MMP2 and MMP9 in multiple myeloma cells

组别	MMP2 mRNA	MMP9 mRNA
空白组	0.979±0.041	1.087±0.101
siRNA-PTEN实验组	2.089±0.155^a	2.670±0.439^a
siRNA对照组	1.149±0.157^b	1.176±0.163^b
F值	63.777	31.007
P值	P<0.001	P<0.001

表4 THP-1诱导分化巨噬细胞中M2巨噬细胞特异性抗体CD163、CD206、F4/80表达水平比较（±s，n=3）

Table 4 Comparison of the expression levels of M2 macrophage-specific antibodies CD163，CD206，and F4/80 induced by THP-1 in differentiated macrophages

组别	CD163+F4/80	CD206+F4/80
M0巨噬细胞组	2.900±0.656	5.567±1.332
瘤细胞上清液组	21.667±0.874^a	11.033±1.498^a
siRNA-PTEN上清液组	46.067±1.872^ab	40.200±1.493^ab
siRNA上清液组	22.300±2.252^ac	12.267±1.858^ac
F值	384.218	299.663
P值	P<0.001	P<0.001

图4 THP-1诱导分化巨噬细胞中M2巨噬细胞特异性抗体CD163、CD206、F4/80表达水平

Figure 4 The expression levels of M2 macrophage-specific antibodies CD163，CD206，and F4/80 induced by THP-1 in differentiated macrophages

表5 THP-1诱导分化巨噬细胞中M2巨噬细胞特异标志物ARG-1、IL-10的mRNA表达水平比较（±s，n=3）

Table 5 Comparison of the expression levels of M2 macrophage-specific markers ARG-1 and IL-10 induced by THP-1 in differentiated macrophages

组别	ARG-1 mRNA	IL-10 mRNA
M0巨噬细胞组	0.965±0.094	1.015±0.099
瘤细胞上清液组	2.208±0.379^a	1.722±0.171^a
siRNA-PTEN上清液组	4.712±1.122^ab	3.653±0.601^ab
siRNA上清液组	2.173±0.298^ac	1.839±0.199^ac
F值	19.826	34.507
P值	P<0.001	P<0.001

参考文献 27

[1]	RAJKUMAR S V, KUMAR S, LONIAL S，et al. Smoldering multiple myeloma current treatment algorithms[J]. Blood Cancer J，2022,12:129. DOI：10.1038/s41408-022-00719-0.
[2]	RAVI G, COSTA L J. Bispecific T-cell engagers for treatment of multiple myeloma[J]. Am J Hematol，2023,98(Suppl 2):S13-21. DOI：10.1002/ajh.26628.
[3]	BRADY R V, THAMM D H. Tumor-associated macrophages：prognostic and therapeutic targets for cancer in humans and dogs[J]. Front Immunol，2023,14:1176807. DOI：10.3389/fimmu.2023.1176807.
[4]	TRUXOVA I, CIBULA D, SPISEK R，et al. Targeting tumor-associated macrophages for successful immunotherapy of ovarian carcinoma[J]. J Immunother Cancer，2023,11(2):e005968. DOI：10.1136/jitc-2022-005968.
[5]	JIANG H F, LI H P. Prognostic values of tumoral MMP2 and MMP9 overexpression in breast cancer：a systematic review and meta-analysis[J]. BMC Cancer，2021,21(1):149. DOI：10.1186/s12885-021-07860-2.
[6]	CHENG Z Y, ZHANG X J, ZHANG Y，et al. Role of MMP-2 and CD147 in kidney fibrosis[J]. Open Life Sci，2022,17(1):1182-1190. DOI：10.1515/biol-2022-0482.
[7]	HASHEMI M, ETEMAD S, REZAEI S，et al. Progress in targeting PTEN/PI3K/Akt axis in glioblastoma therapy：Revisiting molecular interactions[J]. Biomedecine Pharmacother，2023,158:114204. DOI：10.1016/j.biopha.2022.114204.
[8]	中国抗癌协会血液肿瘤专业委员会，中华医学会血液学分会. 中国多发性骨髓瘤骨病诊治指南（2022年版）[J].中华血液学杂志，2022,43(12):979-985.
[9]	段嘉宇，崔健，唐文娇，等. 多发性骨髓瘤疾病全周期健康管理[J]. 中国医学科学院学报，2022,44(4):673-677. DOI：10.3881/j.issn.1000-503X.13467.
[10]	MOREAU P, GARFALL A L, VAN DE DONK N W C J，et al. Teclistamab in relapsed or refractory multiple myeloma[J]. N Engl J Med，2022,387(6):495-505. DOI：10.1056/nejmoa2203478.
[11]	BLADÉ J, BEKSAC M, CAERS J，et al. Extramedullary disease in multiple myeloma：a systematic literature review[J]. Blood Cancer J，2022,12(3):45. DOI：10.1038/s41408-022-00643-3.
[12]	PAUL B, LIEDTKE M, KHOURI J，et al. A phaseⅡ multi-arm study of magrolimab combinations in patients with relapsed/refractory multiple myeloma[J]. Future Oncol，2023,19(1):7-17. DOI：10.2217/fon-2022-0975.
[13]	JASINSKI M, BILINSKI J, BASAK G W. The role of the crosstalk between gut microbiota and immune cells in the pathogenesis and treatment of multiple myeloma[J]. Front Immunol，2022,13:853540. DOI：10.3389/fimmu.2022.853540.
[14]	SUN J, PARK C, GUENTHNER N，et al. Tumor-associated macrophages in multiple myeloma：advances in biology and therapy[J]. J Immunother Cancer，2022,10(4):e003975. DOI：10.1136/jitc-2021-003975.
[15]	WANG S S Y, CHNG W J, LIU H Y，et al. Tumor-associated macrophages and related myelomonocytic cells in the tumor microenvironment of multiple myeloma[J]. Cancers，2022,14(22):5654. DOI：10.3390/cancers14225654.
[16]	KUMARI N, CHOI S H. Tumor-associated macrophages in cancer：recent advancements in cancer nanoimmunotherapies[J]. J Exp Clin Cancer Res，2022,41(1):68. DOI：10.1186/s13046-022-02272-x.
[17]	ARVANITAKIS K, KOLETSA T, MITROULIS I，et al. Tumor-associated macrophages in hepatocellular carcinoma pathogenesis，prognosis and therapy[J]. Cancers：Basel，2022,14(1):226. DOI：10.3390/cancers14010226.
[18]	SALTARELLA I, ALTAMURA C, CAMPANALE C，et al. Anti-angiogenic activity of drugs in multiple myeloma[J]. Cancers：Basel，2023,15(7):1990. DOI：10.3390/cancers15071990.
[19]	XU Z J, CHEN Y, MA L，et al. Role of exosomal non-coding RNAs from tumor cells and tumor-associated macrophages in the tumor microenvironment[J]. Mol Ther，2022,30(10):3133-3154. DOI：10.1016/j.ymthe.2022.01.046.
[20]	LI Y T, CHEN Z M, HAN J H，et al. Functional and therapeutic significance of tumor-associated macrophages in colorectal cancer[J]. Front Oncol，2022,12:781233. DOI：10.3389/fonc.2022.781233.
[21]	PEREZ L G, KEMPSKI J, MCGEE H M，et al. TGF-β signaling in Th17 cells promotes IL-22 production and colitis-associated colon cancer[J]. Nature communications，2020,11(1):2608. DOI：10.1038/s41467-020-16363-w.
[22]	JIA S N, HAN Y B, YANG R，et al. Chemokines in colon cancer progression[J]. Semin Cancer Biol，2022,86(pt 3):400-407. DOI：10.1016/j.semcancer.2022.02.007.
[23]	CHERUKU S, RAO V, PANDEY R，et al. Tumor-associated macrophages employ immunoediting mechanisms in colorectal tumor progression：current research in Macrophage repolarization immunotherapy[J]. Int Immunopharmacol，2023,116:109569. DOI：10.1016/j.intimp.2022.109569.
[24]	ANAGNOSTAKIS F, PIPERI C. Targeting options of tumor-associated macrophages（TAM）activity in gliomas[J]. Curr Neuropharmacol，2023,21(3):457-470. DOI：10.2174/1570159X20666220120120203.
[25]	VAGO J P, AMARAL F A, VAN DE LOO F A J. Resolving inflammation by TAM receptor activation[J]. Pharmacol Ther，2021,227:107893. DOI：10.1016/j.pharmthera.2021.107893.
[26]	DING J L, WANG J, CAI X P，et al. Granulocyte colony-stimulating factor in reproductive-related disease：function，regulation and therapeutic effect[J]. Biomed Pharmacother，2022,150:112903. DOI：10.1016/j.biopha.2022.112903.
[27]	JOHN L, KRAUTH M T, PODAR K，et al. Pathway-directed therapy in multiple myeloma[J]. Cancers，2021,13(7):1668. DOI：10.3390/cancers13071668.

多发性骨髓瘤细胞通过PI3K/AKT信号通路促进M2巨噬细胞极化的机制研究

Study on the Mechanisms of Multiple Myeloma Cells Promoting M2 Macrophage Polarization through PI3K/AKT Signaling Pathway

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