M2型肿瘤相关巨噬细胞在胰腺癌中的表达及其临床意义

目的探讨CD68、CD163标记的M2型肿瘤相关巨噬细胞（Tumor-associated macrophage,TAM）在胰腺癌中的表达及其临床意义。方法应用免疫组织化学检测CD68、CD163蛋白在50例胰腺导管腺癌中的表达;分析它们与临床病理特征之间的关系。结果CD68、CD163阳性表达率分别为84%和76%;它们与癌细胞同时聚集在癌侵袭边缘、血管、淋巴管、神经周围;CD68、CD163阳性表达与患者年龄、性别、肿瘤部位、病理分级无关（P>0.05）,与肿瘤大小、淋巴结转移、血管侵犯和临床分期密切相关（P<0.05）。结论M2型肿瘤相关巨噬细胞在胰腺癌中广泛浸润,它可能促进了胰腺癌的恶性进展。     

Involvement of protumor macrophages in breast cancer progression and characterization of macrophage phenotypes

Tumor-associated macrophages (TAMs) are the most prominent immune cells in the breast cancer microenvironment, and the protumor functions of TAMs are thought to affect cancer progression and resistance to anticancer therapy. Numerous studies using human breast cancer samples, cell lines, and murine breast cancer models have revealed details of the mechanisms by which the protumor functions of TAMs are activated. Recent advances have highlighted the significant involvement of TAMs in the resistance of breast cancer cells to immunotherapy. Tumor-associated macrophages express a number of immunosuppressive genes, and single-cell sequence analyses of human and murine cancer samples have helped elucidate the mechanism of TAM-induced immunosuppression. As TAMs are considered suitable targets for anticancer therapies, we summarized the protumor functions of TAMs and the potential of anticancer therapies targeting TAMs, with a focus on breast cancer research.     

CD68巨噬细胞浸润对原发性食管小细胞癌预后的影响

目的：探讨不同亚型肿瘤相关巨噬细胞在原发性食管小细胞癌(PESC)中的浸润情况及其与患者临床病理指标和预后的关系。方法：回顾性分析汕头大学医学院附属肿瘤医院2000年1月—2019年12月进行手术治疗且临床病理资料完整的69例PESC患者组织标本。采用免疫组织化学法评估CD68巨噬细胞及CD163巨噬细胞在PESC中的浸润情况,卡方检验和Fisher精确检验分析其与PESC临床病理指标的关系,采用Kaplan-Meier生存分析及多因素Cox回归方法分析各检测指标与患者生存预后的关系。结果：CD68及CD163均在巨噬细胞的胞质或胞膜表达。CD68巨噬细胞与肿瘤T分期有关(χ²=6.336,r=-0.303,P=0.012),与其他各临床病理指标均无明显相关。PESC患者的pTNM分期、手术淋巴结清扫数目、N分期、辅助性化疗及CD68巨噬细胞浸润情况与总生存期有关(P<0.05)。多因素Cox回归分析结果显示pTNM分期、手术淋巴结清扫数目、CD68巨噬细胞是PESC患者独立的预后因素。与CD68巨噬细胞低浸润组相比,CD68巨噬细胞高浸润患者的死亡风险降低[HR=0.340,95% CI (0.179,0.645),P=0.001]。结论：CD68巨噬细胞高浸润是PESC患者独立的预后影响因素,CD68巨噬细胞高浸润的患者预后明显优于CD68巨噬细胞低浸润的患者。     

The Distribution of M2 Macrophage and Treg in Nasopharyngeal Carcinoma Tumor Tissue and the Correlation with TNM Status and Clinical Stage

Objective: This study aimed to identify the distribution of M2 macrophage and Treg in Nasopharyngeal Carcinoma (NPC) tumor tissue samples. The presence of these two groups of cells was further correlated to clinical stage, tumor size, the lymphatic node involvement, and metastasis. Methods: The total of 50 formalin-fixed paraffin-embedded (FFPE) NPC tissue samples was collected retrospectively (27 samples) and prospectively (23 samples). Samples were FFPE tissue slices. Immunohistochemistry was done on the FFPE tissue slides using anti-CD-163 and anti-FoxP-3 antibodies for M2 macrophage and Treg detection, respectively. The M2 macrophage interpretation was performed by eye-balling method and the score was divided into 0 (negative), 1 (scant), 2 (focal), and 3 (abundant). The average number of Treg FOXP3+ cells in 5 high power fields (HPF) was calculated. The relationship of M2 macrophage and Treg was tested with Spearman’s correlation. The relationship between M2 macrophage and Treg with clinical stage, tumor size, node involvement and metastasis was tested by chi square, with p<0.1. Results: M2 macrophage and Treg were positive correlated (r=0.469, p<0.001). The presence of M2 macrophage and regulatory T cell (Treg) was significantly correlated to tumor size (p= 0.091 for M2 macrophage and p=0.022 for Treg) and clinical stage (p= 0.030 for M2 macrophage and p= 0.002 for Treg), but did not correlate with lymphatic node involvement and metastasis. Conclusions: In Epstein-Barr virus related NPC tumor microenvironment, the presence of M2 macrophage was correlated with Treg, and both types of the cells were correlated with tumor size and clinical stages.     

肿瘤相关巨噬细胞在胃癌中的研究进展

研究表明,肿瘤的侵袭和转移不仅与肿瘤自身的特性有关,还与肿瘤的微环境关系密切。肿瘤相关巨噬细胞（ Tumor associated macrophage ,TAM）是肿瘤微环境中数量较多的炎性细胞,在不同的信号刺激下巨噬细胞分化成经典型的M1型巨噬细胞或替代激活型的M2型巨噬细胞。在许多恶性肿瘤中,不少研究证实,M2型巨噬细胞参与了肿瘤的侵袭和转移,胃癌的研究得出相似结论。 TAM的促肿瘤作用可能存在凝血因子的参与。缺氧环境亦可能在这一过程中起到重要作用。研究发现,胃癌中TAM的功能随位置的不同而变化,与细胞分化、胃癌分型无关。本文就TAM在胃癌中的研究现状做一综述,以期全面了解TAM在胃癌中的相互作用,为进一步深入研究二者的关系提供线索。     

巨噬细胞游走抑制因子在卵巢癌侵袭中的作用与意义

背景与目的:巨噬细胞游走抑制因子(maerophage migration inhibitory factor,MIF)与肿瘤的恶性进展密切相关.本研究探讨MIF基因对卵巢癌HO-8910和OVCAR-3细胞侵袭和增殖能力的影响及其在卵巢癌组织中表达的意义.方法:瞬时转染小干扰RNA(small interfering RNA,siRNA)靶向敲除MIF基因,RT-PCR和Western blot检测MIF在mRNA和蛋白水平的表达,通过体外迁移、侵袭实验和噻唑蓝比色法(MTT)分析MIF对HO-8910和OVCAR-3细胞迁移、侵袭和增殖能力的影响;免疫组织化学检测MIF蛋白在卵巢癌组织中的表达情况.结果:与阴性对照组细胞比较,瞬时转染MIF siRNA的HO-8910和OVCAR-3细胞中MIF基因表达水平明显降低.MTT结果显示,转染MIF siRNA的两株卵巢癌细胞增殖率显著低于阴性对照组(P＜0.05).转染MIF siRNA的HO-8910细胞穿膜细胞数(MIF-si1:48.0±7.3;MIF-si2:38.0±3.6)与阴性对照组(78.0±8.5)比较差异有统计学意义(P＜0.05),其穿过铺了Matrigel膜的细胞数(MIF-sil:35.0±5.0:MIF-si2:30.0±5.6)也显著少于阴性对照组(P＜0.05).同样,转染了MIF siRNA的OVCAR-3细胞穿膜细胞数(MIF-si1:40.0±4.5;MIF-si2:42.0±3.0)与阴性对照组(65±2.1)比较,差异也有统计学意义(P＜0.05),其穿过铺了Matrigel膜的细胞数(MIF-sil:25.0±3.0:MIF-si2:27.0±3.4)也明显低于阴性对照组(P＜0.05).53.6%(37/69)卵巢癌组织中有MIF蛋白表达.MIF蛋白表达与肿瘤临床分期呈显著正相关(P＜0.01).结论:MIF基因可能通过促进肿瘤细胞的增殖和侵袭能力在卵巢癌的发生发展中发挥重要作用,有可能成为分子靶向治疗卵巢癌的潜在靶点.     

miR-34c-3p与M2型肿瘤相关巨噬细胞在三阴性乳腺癌中的表达及意义

目的:检测miR-34c-3p与M2型肿瘤相关巨噬细胞（tumor-associated macrophage,TAM)在三阴性乳腺癌(triple negative breast cancer,TNBC)中的表达,并探讨其在乳腺癌发病中的意义。方法:选择2017年2月至2018年1月于西京医院就诊的68例TNBC患者作为实验组(A组),以同期就诊的70例乳腺纤维腺瘤患者作为对照组(B组)。采用Real-time RT-PCR 检测组织标本中miR-34c-3p的表达;流式细胞检测M2型TAM的表达;ELISA法检测血清中IL-10的表达。并进一步分析miR-34c-3p的表达与乳腺癌M2型TAM及血清IL-10水平的相关性。结果:A、B组miR-34c-3p的相对表达量分别为0.84±0.08、1.29±0.71,A组明显低于B组,差异有统计学意义(P<0.05)。与B组比较,A组M2型TAM(CD68+CD163+)细胞比例显著升高(P<0.05)。A组血清IL-10的表达［(19.69±4.93) pg/ml］较B组［(17.26±3.51) pg/ml］显著升高,差异有统计学意义(P<0.05)。TNBC组织中miR-34c-3p的表达与M2型TAM比例及血清IL-10表达均呈显著负相关(r=-0.508,r=-0.656,P<0.05)。结论:TNBC组织中miR-34c-3p的表达下调,M2型TAM比例及血清IL-10表达均显著升高,促进TNBC进展。     

CD163+ tumor-associated macrophage is a prognostic biomarker and is associated with therapeutic effect on malignant pleural effusion of lung cancer patients

CD163+ tumor-associated macrophages (TAMs) play an important role in the progression of cancer. However, the significance of CD163+ TAMs in malignant pleural effusion (MPE) is still unclear. The aim of this study is to evaluate the prognostic value of CD163+ TAMs in MPE, and the regulatory effect of an immune adjuvant (pseudomonas aeruginosa - mannose-sensitive hemagglutinin, PA-MSHA, which is used for MPE treatment in clinic) on CD163+ TAMs in MPE. Here, we found that the percentage of CD163+ TAMs in MPE was significantly higher than that in non-malignant pleural effusion (P<0.001). More importantly, CD163+ TAMs in MPE patients were an independent prognostic factor for progression-free survival. M2-related cytokines were highly expressed in MPE-derived CD163+ TAMs than in MPE-derived CD163− macrophages (P<0.05). CD163+ TAMs frequency in MPE patients was obviously reduced after PA-MSHA treatment in clinic (P<0.05). After treatment with PA-MSHA, M2 macrophages were re-educated to M1 macrophages in vitro. TLR4 blocking antibody inhibited M2 macrophages polarization to M1 macrophages induced by PA-MSHA. These findings highlight that accumulation of CD163+ TAMs in MPE caused by lung cancer is closely correlated with poor prognosis. CD163+ TAMs are associated with therapeutic effect in MPE. PA-MSHA re-educates CD163+ TAMs to M1 macrophages through TLR4-mediated pathway in MPE.     

CD44在卵巢上皮性肿瘤中的表达及其临床意义

目的研究黏附分子CD44在卵巢上皮性肿瘤中的表达及其表达与卵巢癌临床特征的关系。方法应用RT-PCR、巢式PCR、光密度扫描和免疫组化等方法,从mRNA和蛋白两个水平检测了卵巢上皮性肿瘤中CD44的表达,并分析了其与临床病理特征的关系。结果CD44标准型(CD44H)在恶性组和良性组中的蛋白表达率分别为61.5%和18.8%,差异有显著性;CD44HmRNA定量分析     

CD44v6基因在宫颈癌组织中的表达及其临床意义

目的:探讨CD44v6基因在宫颈癌组织中的表达情况及其临床意义。方法:用免疫组织化学S-P法,检测56例宫鳞癌,32例宫颈腺癌中CD44v6蛋白的表达情况并分析相关的临床病理因素。结果:CD44v6表达与宫颈癌组织学类型无关;无论宫颈腺癌或鳞癌,淋巴结转移组中CD44v6阳性表达明显高于无淋巴结转移组（P<0.05）;CD44v6表达与宫颈癌临床分期、病理分级、病灶大小无关（P>0.05）。结论:CD44v6在宫颈癌淋巴结转移及癌细胞分化过程中可能起着重要作用,对早期预测癌细胞的转移潜能、判断宫颈癌预后具有一定的临床意义。     

在恶性肿瘤患者中检测蛋白质Z的临床意义

背景与目的:恶性肿瘤患者常存在血液高凝状态,其高凝状态与肿瘤转移的关系越来越受到人们的重视。蛋白质Z(proteinZ,PZ)是一种新发现的抗凝因子,是由肝脏合成的维生素K依赖的血浆蛋白,其结构与维生素K依赖性凝血因子FⅦ、FⅨ、FⅩ及蛋白质C分子十分相似,但其生理功能及临床意义尚不十分明确。临床研究发现PZ水平变化与出、凝血异常导致的疾病危险程度相关。但对实体肿瘤患者PZ水平变化报道很少。本研究重点在探讨恶性肿瘤患者PZ与凝血因子FⅦ∶C、FⅨ∶C、FⅩ∶C、FⅩ∶Ag水平变化的关系及其临床意义。方法:对80例恶性肿瘤患者血浆PZ、FⅦ∶C、FⅨ∶C、FⅩ∶C、FⅩ∶Ag进行测定分析,并作相关性分析;以80名健康人的检测结果作为对照。结果:肿瘤患者的PZ值(1210.89±251.13)滋g/L明显低于健康对照组(2378.83±429.51)滋g/L(P<0.01),但恶性肿瘤组的FⅦ∶C、FⅩ∶C、FⅩ∶Ag水平明显较对照组升高,两组分别为(162.42±36.57)%、(120.27±33.96)%、(133.66±35.51)%和(114.78±28.96)%、(79.23±19.46)%、(93.00±17.73)%(P<0.01),肿瘤患者FⅨ∶C水平(119.86±56.39)%与健康对照者(109.21±36.46)%之间的差异无显著性(P>0.05)。相关性分析发现恶性肿瘤组及对照组中的PZ水平与FⅦ∶C、FⅩ∶C、FⅩ∶Ag水平之间均存在明显负相关(P<0.01),而PZ与FⅨ∶C无显著性相关(P>0.05)。分析肿瘤病程与上述凝血因子的关系发现,Ⅲ～Ⅳ期患者(局部晚期和晚期)PZ水平较Ⅱ期患者降低更明显,分别为(998.32±117.72)ng/ml与(1326.29±245.7)ng/ml(P<0.01);而其FⅦ∶C、FⅩ∶C、FⅩ∶Ag水平比Ⅱ期患者更高[(206.76±28.63)%、(162.53±32.92)%、(168.03±25.97)%vs.(136.09±26.80)%、(101.89±23.44)%、(105.41±13.86)%,P<0.01]。结论:PZ水平在恶性肿瘤患者中明显降低,PZ与FⅦ∶C、FⅩ∶C、FⅩ∶Ag存在明显负相关;PZ水平随着肿瘤病程进展而下降更明显,可能是恶性肿瘤预后不良因素之一。     

High CYP2E1 activity aggravates hepatofibrosis by limiting macrophage polarization towards the M2 phenotype

Cytochrome P450 2E1 (CYP2E1) is an important drug‐metabolizing enzyme that has been recognized as one of the risk factors for hepatofibrosis. Macrophages play key roles in regulating hepatofibrosis progression and resolution. However, whether CYP2E1 is involved in the regulation of macrophage polarization during hepatofibrosis is still unclear. Herein, we measured CYP2E1 activity and the expression of CD163 (an M2 marker) and CD68 (a pan‐macrophage marker) in hepatofibrotic tissue from HCC patients (n = 26) with comparison to normal liver tissue (n = 26). The relationship of CYP2E1 activity in vivo and the CD163/CD68 ratio (an indicator of M2 polarization), as well as the extent of hepatofibrosis, were evaluated in diethylnitrosamine (DEN)‐treated Sprague‐Dawley rats. Strikingly, CYP2E1 activity and expression of CD68 increased and the CD163/CD68 ratio decreased, especially in hepatofibrotic tissue with higher CYP2E1 activity. Expression of α‐SMA, Ki67, and PCNA were positively correlated with CYP2E1 activity and inversely correlated with the CD163/CD68 ratio. Furthermore, CYP2E1 activity showed an inverse correlation with the CD163/CD68 ratio. Overall, high CYP2E1 activity aggravates hepatofibrosis by restraining M2 macrophage polarization, providing a novel insight for understanding the profibrotic activity of CYP2E1 and a promising avenue for hepatofibrosis therapy.     

人胃癌组织中巨噬细胞移动抑制因子和VEGF的表达及其临床意义

目的：检测巨噬细胞移动抑制因子（macrophage migration inhibitory factor, MIF）和VEGF在胃癌组织中的表达,分析其与胃癌临床病理特征之间的关系。方法：选取河南大学淮河医院2008—2013年手术切除的89例胃癌组织与2013年手术切除的新鲜胃癌及癌旁标本4对,分别采用免疫组织化学法和Western boltting方法检测胃癌组织及其癌旁组织中MIF和VEGF的表达,结合临床资料分析其表达水平与胃癌临床病理特征之间的关系。结果：MIF和VEGF在胃癌中的表达率分别为88.8%和50.6%,而在癌旁组织中均无表达。胃癌组织中MIF（0.87±0.29 vs 0.23±0.14,P<0.05）和VEGF（0.89±023 vs 0.34±0.21,P<0.05）相对表达量均显著高于癌旁组织。MIF和VEGF表达水平在不同年龄、性别组间无显著差异,而在不同分化程度和TNM临床分期组间差异有统计学意义（P<0.05）。结论：MIF和VEGF在胃癌组织中呈高表达,且可能与胃癌的分化程度和TNM临床分期有关。     

Prostate cancer-derived CCN3 induces M2 macrophage infiltration and contributes to angiogenesis in prostate cancer microenvironment

Tumor-associated macrophages (TAMs) are M2-polarized macrophages that infiltrate the tumor microenvironment and promote tumorigenesis. However, the mechanisms by which TAMs modulate prostate cancer (PCa) growth are poorly understood. Here, we found that expression of Nephroblastoma Overexpressed (NOV/CCN3) is upregulated in PCa cells and correlated with M2 macrophage infiltration. RAW264.7 macrophage migration was induced by conditioned media (CM) from various PCa cells in proportion to the cellular level of CCN3 expression and was inhibited by an anti-CCN3 neutralizing antibody. CCN3 and PCaCM treatment skewed RAW264.7 cell differentiation from an M1 phenotype to an M2 phenotype. PCa-derived CCN3 induced focal adhesion kinase (FAK)/Akt/NF-κB signaling in RAW264.7 cells, which resulted in VEGF expression and subsequently increased tube formation in endothelial progenitor cells. Finally, PCa-secreted CCN3 stimulated RAW264.7 cells and promoted angiogenesis in the chick chorioallantoic membrane assay (CAM), and increased tumor growth and tumor-associated angiogenesis in a PCa xenograft mouse model. Our results indicate that PCa-secreted CCN3 can recruit macrophages and skew their differentiation to an M2 phenotype. In turn, CCN3-stimulated macrophages contribute to VEGF-dependent angiogenesis. This study reveals a novel mechanism by which TAMs enhance PCa angiogenesis and identifies a potential therapeutic target for PCa.     

Development and experimental validation of an M2 macrophage and platelet-associated gene signature to predict prognosis and immunotherapy sensitivity in bladder cancer

Platelets and M2 macrophages both play crucial roles in tumorigenesis, but their relationship and the prognosis value of the relative genes in bladder cancer (BLCA) remain obscure. In the present study, we found that platelets stimulated by BLCA cell lines could promote M2 macrophage polarization, and platelets were significantly associated with the infiltration of M2 macrophages in BLCA samples. Through the bioinformatic analyses, A2M, TGFB3, and MYLK, which were associated with platelets and M2 macrophages, were identified and verified in vitro and then included in the predictive model. A platelet and M2 macrophage-related gene signature was constructed to evaluate the prognosis and immunotherapeutic sensitivity, helping to guide personalized treatment and to disclose the underlying mechanisms.     

人巨噬细胞金属弹性蛋白酶在胃癌细胞和组织中的表达及临床意义

目的测定人巨噬细胞金属弹性蛋白酶(HME)在胃癌细胞及胃癌组织中的表达,评价其在胃癌进展中的作用。方法采用实时荧光PCR测定胃癌细胞及胃癌组织中HME mRNA表达水平,并对胃癌组织中HME mRNA表达与患者临床病理特征及预后的关系进行统计分析。结果MGC-803、SGC-7901和AGS胃癌细胞株均有HME mRNA的表达。胃癌组织、癌旁组织和正常组织的HME mRNA表达阳性率分别为31.03%、25.87%和18.97%,阳性结果拷贝数对数值分别为6.06±0.76、3.53±0.85和1.88±0.82,胃癌组织HME tuRNA表达阳性率、HME mRNA表达量均显著高于正常组织(P<0.05)。HME mRNA表达与肿瘤的位置、大小、浸润深度、耐药基因topⅡ和PG的表达无关,胃癌组织HME mRNA阳性患者淋巴结转移发生率低于阴性者、GST基因表达阳性率低于阴性者、2年生存率高于阴性者。结论胃癌组织中HME mRNA表达阳性与胃癌转移潜能降低有关,可能是预后良好的标志。     

晚期恶性肿瘤血清VEGF含量测定的临床意义

Objective: To elucidate the clinical significance of serum vascular endothelial growth factor (VEGF) level in patients with advanced cancer. Methods: Enzyme linked immunosorbent assay (ELISA) was used to determine the serum VEGF concentration in 40 patients with advanced cancer [non-small cell lung cancer (NSCLC), esophageal cancer (EC) and nasopharyngeal carcinoma (NPC)] before and after chemotherapy and 10 healthy volunteers as control group. Results: The serum VEGF concentrations in 40 cases of advanced cancer patients were significantly higher than those of 10 healthy control cases [(477.07 ± 374.10 ) pg/mL vs (139.09 ± 133.41 ) pg/mL; P = 0.016]. The serum VEGF concentrations in patients with NSCLC, EC and NPC were (518.53 ± 378.99) pg/mL, (399.21 ± 393.69) pg/mL and (500.68 ± 348.48) pg/mL, respectively. The differences were all statistically significant as compared with healthy control group (P values were 0.011,0.044 and 0.019, respectively). The serum VEGF concentrations of the patients in response to chemotherapy was significantly lower than those of the same patients before they undergoing chemotherapy [(400.41 332.84) pg/mL vs (777.10 ± 666.01) pg/mL; P = 0.034]. Conclusion: The serum VEGF level might be a novel and promising tumor marker of advanced malignancies and a predictor of disease progression, prognosis and therapeutic efficacy.     

晚期恶性肿瘤血清VEGF含量测定的临床意义

目的：探讨血清血管内皮生长因子（VEGF）浓度在晚期恶性肿瘤中的临床意义。方法：应用酶联免疫吸附试验（ELISA）测定40例晚期恶性肿瘤（非小细胞肺癌、鼻咽癌、食管癌）患者血清的VEGF浓度,10名健康成人作为对照。结果：40例晚期恶性肿瘤患者血清VEGF浓度为（477．07±374．10）pg／ml,显著高于健康成人（139．09±133．41）pg／ml,差异有统计学意义（P=0．016）,其中治疗前血清VEGF浓度在非小细胞肺癌为（518．53±378．99）pg／ml,食管癌为（399．21±393．69）pg／ml,鼻咽癌为（500．68±348．48）pg／ml,与健康成人比较差异有统计学意义（P值分别为0．011、0．044和0．019）。化疗有效患者的血清VEGF浓度（400．41±332．84）pg／ml显著低于化疗前浓度（777．10±666．01）pg／ml,差异有统计学意义（P=0．034）。结论：血清VEGF可作为晚期恶性肿瘤监测病情、判断放疗和预后一个有用的指标。