髓系来源的抑制性细胞在小鼠H22原位种植性肝癌中的表达及其意义

目的:比较小鼠H22原位种植性肝癌模型中髓系来源的抑制性细胞（MDSCs）和调节性T细胞（Tregs）的表达,确定主要的免疫抑制性细胞。并探讨主要免疫抑制性细胞在小鼠H22原位种植性肝癌模型中的意义。方法:建立小鼠H22原位种植性肝癌模型,用流式细胞术的方法比较外周血、脾脏、肿瘤组织中MDSCs、Tregs的比例。分析MDSCs与CD4+T细胞、CD8+T细胞、NK细胞、NKT细胞的相关性。通过脾脏切除的方法下调MDSCs的表达,观察对肿瘤存活的影响。结果:MDSCs的比例无论在外周血、脾脏还是肿瘤组织中都显著高于Tregs。MDSCs的比例与CD4+T细胞、CD8+T细胞、NK细胞、NKT细胞呈负相关。下调MDSCs的比例可以显著延长小鼠存活期。结论:小鼠H22原位种植性肝癌中主要的免疫抑制性细胞是MDSCs。MDSCs的高表达预示着不良的小鼠存活期。     

髓系抑制性细胞与肿瘤

髓系抑制性细胞(MDSC)是一群髓系来源具有抑制功能的天然免疫细胞,在肿瘤进展中发挥负向免疫调控作用。MDSC具有强大的抑制功能及显著的异质性,通过多种机制调控固有免疫及适应性免疫系统,发挥促肿瘤作用,同时可通过非免疫机制促进肿瘤血管生成及肿瘤转移等。近年来对其分化、增殖、抑制功能等的研究日趋成熟,由此衍生的靶向针对MDSC的肿瘤免疫治疗研究将为肿瘤疫苗的增效及肿瘤的治疗等带来新的希望。     

髓系抑制性细胞与肿瘤

髓系抑制性细胞(MDSC)是一群髓系来源具有抑制功能的天然免疫细胞,在肿瘤进展中发挥负向免疫调控作用.MDSC具有强大的抑制功能及显著的异质性,通过多种机制调控固有免疫及适应性免疫系统,发挥促肿瘤作用,同时可通过非免疫机制促进肿瘤血管生成及肿瘤转移等.近年来对其分化、增殖、抑制功能等的研究日趋成熟,由此衍生的靶向针对MDSC的肿瘤免疫治疗研究将为肿瘤疫苗的增效及肿瘤的治疗等带来新的希望.     

肿瘤相关免疫抑制细胞与免疫逃逸誊几制研究进展

肿瘤相关树突细胞，髓系来源抑制细胞和肿瘤相关巨噬细胞等在肿瘤免疫逃逸过程中能够诱导免疫抑制反应。肿瘤源性因子可以影响其分化、功能和迁移。细胞内信号转导途径的激活可以调节细胞内代谢、细胞因子的产生和共刺激及共抑制分子的表达，由此产生免疫抑制作用。这些方面的研究将会为抗肿瘤免疫治疗提供新的策略。     

三氧化二砷治疗慢性粒细胞白血病25例

 【摘要】　目的　观察三氧化二砷（As2O3）治疗成年人慢性粒细胞性白血病（CML）的疗效。方法　对25例成年CML患者采用As2O3进行治疗。其中初治18例,复治7例。结果　25例中获骨髓完全缓解9例（36 ％）,部分缓解12例（48 ％）,未缓解4例（16 ％）,总缓解率（有效率）为84 ％。不良反应主要有水肿、皮疹、胃肠道反应,肝功能损害为一过性的氨基转移酶升高。结论　As2O3为治疗CML的一种有效药物,且不良反应较轻,患者耐受性好。     

靶向髓系抑制性细胞的药物在肿瘤治疗中的研究进展

髓系抑制性细胞(myeloid-derived suppressor cells,MDSCs)是机体在肿瘤等病理情况下髓系细胞分化发育受阻而形成的一群未成熟的髓系细胞。肿瘤微环境中的MDSCs具有较强的免疫抑制功能,并与肿瘤的发展密切相关,寻找以MDSCs为靶点的药物在肿瘤治疗中具有良好的应用前景。近年来,许多研究发现多种药物通过促进MDSCs分化、或抑制MDSCs扩增、或诱导MDSCs凋亡或减弱MDSCs的免疫抑制功能等方式抑制肿瘤的进展,这些研究为肿瘤的临床治疗提供了新策略。     

三氧化二砷诱导恶性黑色素瘤A375和B16细胞凋亡的研究

目的: 研究不同浓度的三氧化二砷(As-2O-3)对恶性黑色素瘤人A-{375}细胞和小鼠B-{16}细胞的凋亡诱导作用,为As-2O-3治疗恶性黑色素瘤提供新的理论和实验依据.方法:用流式细胞术(FCM)检测A-{375}细胞和B-{16}细胞的凋亡诱导及杀伤作用:用不同剂量的As-2O-3(0.1 mmol*L{-1}、0.25 mmol*L{-1}和0.5 mmol/L{-1}),对小鼠进行腹腔注射(10 ml*kg{-1},连续注射10 d后,检测小鼠血清与心、肝、肾有关的生化指标.结果:As-2O-3浓度分别为5 μmol*L{-1}、10 μmol*L{-1}和20 μmol*L{-1}时,A-{375}和B-{16}细胞的凋亡率分别为11.85 %、55.51 %、54.90 %和9.81 %、26.45 %、9.93 %:As-2O-3诱导A-{375}和B-{16}细胞总的凋亡和坏死率分别为11.90 %、55.71 %、57.40 %和12.96 %、26.99 %、87.13 %;不同剂量的As-2O-3(0.1 mmol*L{-1}～0.5 mmol*L{-1})对小鼠肝、肾和心肌等功能无明显影响,与对照组各项指标无显著性差异(P>0.05).结论:不同浓度的As-2O-3能明显诱导恶性黑色素瘤A-{375}及B-{16}细胞凋亡和坏死,对A-{375}细胞的凋亡诱导作用强于B-{16}细胞.As-2O-3(0.1 mmol*L{-1}～0.5 mmol*L{-1})对小鼠肝、肾和心肌等功能无明显损伤.     

三氧化二砷抑制恶性黑色素瘤B_(16)细胞生长及对端粒酶活性的影响

背景与目的:研究三氧化二砷(As2O3)对恶性黑色素瘤B16细胞生长抑制作用及对端粒酶活性表达的影响,探讨砷及其化合物治疗恶性黑色素瘤的作用机制,为临床治疗恶性黑色素瘤提供新的理论和实验依据。材料与方法:四甲基噻唑氮蓝(Methylthiazolyltetrazolium,MTT)比色法检测As2O3对B16细胞的生长抑制作用:端粒重序列扩增酶联免吸附实验(Telomericrepeatamplificationprotocolenzyme_linkedimmunosorbantassay,TRAR_ELISA)和聚内烯酰胺凝胶电泳银染(Telomericrepeatamplificationprotocol,polyacrylamidegelelectrophoresissilver_staining,TRAP_PAGE)法检测B16细胞的端粒酶活性。结果:As2O3可显著抑制B16细胞的生长并可下调端粒酶活性,其抑制作用有显著的时间和剂量依赖关系。结论:As2O3对恶性黑色素瘤B16细胞生长抑制作用及对端粒酶活性表达的抑制作用随药物浓度的增加和作用时间的延长而增强。     

髓源抑制性细胞的功能及其逆转策略

髓源抑制性细胞（myeloid-derived suppressor cell,MDSC）是在骨髓中产生的一群具有高度异质性的免疫抑制细胞, 在 肿瘤等病理状态下大量聚集,是促进肿瘤进展、降低患者对传统治疗反应性的关键因素。近年来,免疫检查点阻断剂和基因工程 T细胞过继回输治疗延长了许多晚期恶性肿瘤患者的生存期,但上述免疫疗法在肺癌、结直肠癌等实体瘤中有效率仅为 15%~40%,这与实体瘤免疫抑制微环境密切相关。MDSC在肿瘤微环境中聚集,通过抑制T细胞或NK细胞增殖及功能减弱宿主 抗肿瘤免疫反应,是患者对免疫治疗耐受的关键机制。因此,明确MDSC聚集及功能特征是探索提高免疫治疗效果的重要研究 方向。本文将系统阐述MDSC的产生、聚集及其免疫抑制功能的调控机制,概述目前靶向MDSC治疗的最新研究进展。     

髓源性抑制细胞在肿瘤发生发展中的作用

髓源性抑制细胞(myeloid-derived suppressor cells,MDSCs)是一组异质细胞,在肿瘤相关免疫抑制中起关键作用.MDSCs通过免疫抑制作用,使肿瘤逃避免疫监控.肿瘤组织中MDSCs的浸润与患者预后不良及治疗的抵抗密切相关.MDSCs在转移中发挥重要作用,但MDSCs在远处器官建立转移前微环...     

荷乳腺癌小鼠髓源抑制性细胞对B细胞功能的影响

目的：探讨荷乳腺癌小鼠髓源抑制性细胞（myeloid-derived suppressor cell,MDSC）对B细胞功能的影响。方法：构建BABL/c 小鼠4T1 乳腺癌模型,磁珠分选出荷瘤小鼠脾脏的MDSC与正常小鼠脾脏的B细胞,将MDSC与B细胞共孵育后流式细胞术检测MDSC对B细胞表面分子PD-1、PD-L1、CTLA-4、CCR6、CD62L 和MHCⅡ表达的影响,ELISA 法检测B细胞分泌的IgA、IgM和IgG的变化;BrdU试剂盒检测B细胞增殖情况;Annexin Ⅴ/PI 凋亡试剂盒检测B细胞凋亡。磁珠分选出共孵育体系中的B细胞,将其与T细胞共孵育,BrdU 试剂盒检测T细胞增殖情况,Annexin Ⅴ/PI 凋亡试剂盒检测T细胞凋亡。结果：与B细胞对照组相比,B+MDSC组中B细胞表面PD-L1 表达升高（P<0.01）,PD-1、CTLA-4、CCR6、CD62L 和MHCⅡ的表达均降低(均P<0.01);B细胞分泌的IgA、IgM 和IgG 明显升高（均P<0.01）,B细胞增殖增高（P<0.01）、凋亡降低（P<0.01）。与T细胞对照组相比,B+MDSC (1∶5）+T组的T细胞增殖明显降低（P<0.01）,T细胞凋亡无明显变化。结论：荷乳腺癌小鼠MDSC促进B细胞增殖和抑制B细胞凋亡,并且MDSC诱导的B细胞可以抑制T细胞的增殖。     

三氧化二砷改善胃癌患者术后辅助化疗疗效同时减轻骨髓抑制

Objective: The aim of the study was to investigate the prospective study if treatment with arsenic trioxide (As2O3) could enhance disease-free survival as adjuvant post-operative chemotherapy for gastric cancer patients and protect bone marrow from the negative effects of chemotherapy. Methods: 84 adults were randomized into two groups. Patients in treat- ment group were treated with As203 and FOLFOX regimen, the other were administered with FOLFOX regimen only. Results: Four patients were withdrawn in treatment group after 3-4 cycles and the reasons were headache and fidgety (n = 2), ar- rhythmia (n = 1) and AST/ALT elevation (n = 1), while 1 patient in control group after 4 cycles for neutropenia. In the treatment group, the median DFS was 28.34 months (95% CI, 25-33 months). While in control group, the median DFS was 24.50 months (95% CI, 20-30 months). This difference was not statistically significant (chi-square: 2.8885; P value: 0.0892). Pa- tients in the same subgroup of node-positive was 29 in the treatment group and 32 in control group, respectively. The median DFS was 27.87 months (95% CI, 25-31 months) in the treatment group and 24.18 months (95% CI, 19-31 months) in the control group with promising statistical significance (HR 1.89; chi-square: 4.78; P value: 0.0287). The most common grades 3-4 toxicity was leucopenia (n = 11) in control group and the difference was significant (chi-square: 3.9768, P value: 0.046) compared with that in treatment group (n = 4). Conclusion: The combination of arsenic trioxide and FOLFOX regimen has a potential advantage of enhancing disease-free survival in patients with gastric cancer in nodal-positive status as post-opera- tive chemotherapy, and protect bone marrow from the negative effects of chemotherapy.     

三氧化二砷诱导Raji细胞凋亡与Survivin基因关系的研究

Objective:To investigate the apoptosis induction by arsenic trioxide (As2O3) in Raji cells and its correlation with cell cycle arrest and expression of the Survivin gene.Methods:After Raji cells were treated with As2O3 in different concentrations (1,2,4 and 8 μM),for 24,48 and 72 h,respectively,and cell proliferation was tested by MTT assay.Apoptosis was observed with electron microscope end DNA electrophoresis.The distribution of cell cycles and cell apoptosis were detected by flow cytometry.Expression of the Survivin gene was determined by real-time quantitative RT-PCR.Results:As2O3 (1-8 μM) inhibited Raji cells growth effectively in a dose- and time-dependent manner.As2O3 at 2-8μM could induce cell apoptosis and cell cycle arrest.However,As2O3 (1 μM) inhibited Raji proliferation only by cell cycle arrest,without any symptoms of cell apoptosis.At the same time,Survivin gene expression was down-regulated after the treatment.Conclusion:As2O3 could induce substantial proliferation inhibition,cell cycle arrest and apoptosis in Raji cell.Cell cycle arrest might be a reason why apoptosis occurs.As2O3 can markedly down-regulate expression of the Survivin gene in a dose- and timedependent manner.The down-regulated Survivin gene might be leading to cell apoptosis by As2O3.     

A mutual activation loop between breast cancer cells and myeloid-derived suppressor cells facilitates spontaneous metastasis through IL-6 trans-signaling in a murine model

Introduction

Tumor cell interactions with the microenvironment, especially those of bone-marrow-derived myeloid cells, are important in various aspects of tumor metastasis. Myeloid-derived suppressor cells (MDSCs) have been suggested to constitute tumor-favoring microenvironments. In this study, we elucidated a novel mechanism by which the MDSCs can mediate spontaneous distant metastasis of breast cancer cells.

Methods

Murine breast cancer cells, 4T1 and EMT6, were orthotopically grafted into the mammary fat pads of syngeneic BALB/c mice. CD11b⁺Gr-1⁺ MDSCs in the spleen, liver, lung and primary tumor mass were analyzed. To evaluate the role of MDSCs in the distant metastasis, MDSCs were depleted or reconstituted in tumor-bearing mice. To evaluate whether MDSCs in the metastasizing tumor microenvironment affect breast cancer cell behavior, MDSCs and cancer cells were co-cultivated. To investigate the role of MDSCs in in vivo metastasis, we blocked the interactions between MDSCs and cancer cells.

Results

Using a murine breast cancer cell model, we showed that murine breast cancer cells with high IL-6 expression recruited more MDSCs and that the metastasizing capacity of cancer cells paralleled MDSC recruitment in tumor-bearing mice. Metastasizing, but not non-metastasizing, tumor-derived factors induced MDSCs to increase IL-6 production and full activation of recruited MDSCs occurred in the primary tumor site and metastatic organ in the vicinity of metastasizing cancer cells, but not in lymphoid organs. In addition, tumor-expanded MDSCs expressed Adam-family proteases, which facilitated shedding of IL-6 receptor, thereby contributing to breast cancer cell invasiveness and distant metastasis through IL-6 trans-signaling. The critical role of IL-6 trans-signaling was confirmed in both the afferent and efferent pathways of metastasis.

Conclusion

In this study, we showed that metastasizing cancer cells induced higher MDSCs infiltration and prompted them to secret exaggerated IL-6 as well as soluble IL-6Rα, which, in turn, triggered a persistent increase of pSTAT3 in tumor cells. This potential tumor-MDSC axis involving IL-6 trans-signaling directly affected breast cancer cell aggressiveness, leading to spontaneous metastasis.     

三氧化二砷辅助治疗难治或复发急髓白血病的疗效观察

目的:研究三氧化二砷（As2O3）联合小剂量高三尖杉酯碱（homoharringtonine,HHT）、阿糖胞苷（Ara-c）治疗难治或复发性急性髓细胞白血病（relapsed acute myeloid leukemia,RAML）的疗效及不良反应。方法:难治或复发性急性髓细胞白血病患者180例随机分成两组:研究组94例,方案:三氧化二砷10mg/日,d1-5、d8-12静脉滴注,高三尖杉酯碱1mg/日,d1-14静脉滴注,阿糖胞苷25mg,皮下注射2次/日,d1-14。对照组86例,给予标准剂量HA方案:用药剂量疗程同前,不加As2O3。 结果:研究组68例获得完全缓解（CR）,CR率72.3％（68/94）,13例获得部分缓解（PR）,总有效率（ORR）86.2％（81/94）。对照组CR率54.7％（47/86）,9例PR,总有效率65.1％。两组比较研究组完全缓解率、部分缓解率以及总有效率均优于对照组（P＜0.05）,不良反应发生率研究组与对照组无显著差异（P＞0.05）。结论:小剂量阿糖胞苷方案治疗难治复发性AML的疗效比标准剂量HA方案疗效优越,化疗相关不良反应无显著差异。     

Cysteine cathepsin activity suppresses osteoclastogenesis of myeloid-derived suppressor cells in breast cancer

Cysteine cathepsin proteases contribute to many normal cellular functions, and their aberrant activity within various cell types can contribute to many diseases, including breast cancer. It is now well accepted that cathepsin proteases have numerous cell-specific functions within the tumor microenvironment that function to promote tumor growth and invasion, such that they may be valid targets for anti-metastatic therapeutic approaches. Using activity-based probes, we have examined the activity and expression of cysteine cathepsins in a mouse model of breast cancer metastasis to bone. In mice bearing highly metastatic tumors, we detected abundant cysteine cathepsin expression and activity in myeloid-derived suppressor cells (MDSCs). These immature immune cells have known metastasis-promoting roles, including immunosuppression and osteoclastogenesis, and we assessed the contribution of cysteine cathepsins to these functions. Blocking cysteine cathepsin activity with multiple small-molecule inhibitors resulted in enhanced differentiation of multinucleated osteoclasts. This highlights a potential role for cysteine cathepsin activity in suppressing the fusion of osteoclast precursor cells. In support of this hypothesis, we found that expression and activity of key cysteine cathepsins were downregulated during MDSC-osteoclast differentiation. Another cysteine protease, legumain, also inhibits osteoclastogenesis, in part through modulation of cathepsin L activity. Together, these data suggest that cysteine protease inhibition is associated with enhanced osteoclastogenesis, a process that has been implicated in bone metastasis.     

三氧化二砷对人子宫内膜癌细胞裸鼠模型中Caspase-3表达的影响

目的 通过建立人子宫内膜癌细胞HEC-1-B裸鼠模型,检测不同浓度三氧化二砷(As2 O3)对移植瘤细胞Caspase-3表达的影响,并探讨As2 O3的作用机制.方法 建立人子宫内膜癌HEC-1-B细胞皮下移植瘤模型,按编号随机分为实验组即As2O3低(1.0 mg/kg)、中(2.5 mg/kg)、高(5.0 mg/kg)剂量组、空白对照生理盐水(NS)组及阳性对照顺铂组(5.0 mg/kg),连续腹腔给药8天后,通过流式细胞仪及实时荧光定量RT-PCR检测移植瘤内Caspase-3相对表达量.结果 与生理盐水组比较,低、中浓度As2O3及顺铂均可上调Caspase-3的表达(P<0.05),但高浓度组上调不明显(P>0.05).与顺铂组比较,As2O3低剂量组作用最明显,差异有统计学意义(P<0.05);但与中、高浓度组比较,差异无统计学意义.结论 低浓度As2 O3能上调人子宫内膜癌细胞内Capase-3的表达,且优于顺铂,可能是三氧化二砷诱导细胞凋亡的作用机制之一.