真性红细胞增多症患者JAK2基因突变的临床意义

真性红细胞增多症（polycythemia vera,PV）为一种以红细胞增多为特征的骨髓增殖性肿瘤（myeloproliferative neoplasm,MPN）。PV的发病机制尚未阐明,可能和JAK2基因的功能获得性体细胞突变相关,JAK2基因突变为PV提供了分子学诊断标准。JAK2基因突变和等位基因突变负荷可推测PV患者的临床特征和发展趋势。JAK2基因突变的发现促进了分子靶向治疗的发展,JAK2抑制剂ruxolitinib已应用于临床,治疗效果和安全性均良好。JAK2等位基因突变负荷与白细胞增多、转化为骨髓纤维化（myelofibrosis,MF）密切相关,高JAK2等位基因突变负荷可能为PV患者预后不良的危险因素。本文对PV患者JAK2基因突变的临床意义进行综述。      

JAK1/2抑制剂治疗骨髓增殖性肿瘤的研究进展

bcr-abl阴性的骨髓增殖性肿瘤(MPN)包括真性红细胞增多症(PV)、原发性血小板增多症(ET)和原发性骨髓纤维化(PMF).随着JAK2 V617F基因突变在MPN患者中的发现,一系列针对该突变的小分子靶向药物被研发,其中通过COMFORT-Ⅰ和COMFORT-Ⅱ等试验研究的JAK1/2抑制剂芦可替尼(ruxolitinib)已经被美国食品和药品管理局(FDA)和欧洲药品管理局(EMA)批准应用于中晚期PMF患者和羟基脲耐药或不耐受PV患者的治疗,给MPN患者带来了新的希望.     

紫铆花素抑制脂多糖诱导的骨髓源性巨噬细胞炎症反应的研究

目的：研究紫铆花素（butein）对脂多糖（LPS）诱导小鼠骨髓源性巨噬细胞炎症反应的抑制效应，并探讨JAK2-STAT3通路在其中的作用。方法：分离C57BL/6小鼠骨髓，用40 ng/mL的巨噬细胞集落刺激因子刺激7 d，诱导为骨髓源性巨噬细胞（BMDM）。采用500 ng/mL的LPS刺激BMDM 12 h建立炎症模型，butein干预组采用5、10、20 μmol/L butein与LPS共处理，butein单独处理组为20 μmol/L的butein处理12 h，并设立空白对照组。ELISA法检测BMDM培养液中TNF-α、IL-6和NO的水平；流式细胞术检测细胞内ROS水平；Western blot法检测细胞内iNOS、p-JAK2、JAK2、p-STAT3和STAT3蛋白表达水平，并分析P-JAK2/JAK2和P-STAT3/STAT3蛋白表达水平的比值变化。结果：ELISA实验结果显示，LPS刺激BMDM后，培养液中的TNF-α、IL-6和NO含量显著升高，而5、10、20 μmol/L的butein干预可抑制上述促炎因子的分泌，且呈现剂量-效应关系。流式细胞术检测结果显示，butein抑制了LPS活化BMDM的ROS水平升高。Western blot检测结果表明，LPS刺激后BMDM内iNOS蛋白表达水平升高，JAK2和STAT3蛋白表达水平虽未明显变化，但磷酸化水平显著增加。而butein干预可有效抑制JAK2和STAT3蛋白磷酸化。结论：Butein可抑制LPS诱导的巨噬细胞炎症反应，JAK2-STAT3信号通路可能参与调控这一效应，提示butein是一种炎症相关疾病的候选药物。     

原发性血小板增多症、真性红细胞增多症和骨髓纤维化：目前的处理和靶向治疗

最近发现JAK2和／或MPL突变对真性红细胞增多症（PV）、原发性血小板增多症（ET）和原发性骨髓纤维化（PMF）的诊断和治疗产生着重大影响。例如,JAK2突变目前认为是诊断PV的必要条件,WHO分类系统最近修订的PV、ET和PMF诊断标准中包括JAK2和MPL（血小板生成素受体）突变作为克隆性标记。从治疗学的观点,JAK—STAT信号途径现已确定为研发骨髓增殖性新生物治疗新药的合理靶途径。本文简述ET、PV和PMF目前的处理,并复习抗JAK2小分子药物临床前和临床活性的有关资料。     

抗CD38单克隆抗体治疗多发性骨髓瘤研究进展

多发性骨髓瘤（multiple myeloma，MM）是一种血液系统恶性肿瘤，以骨髓内浆细胞克隆性增殖为特征，发病率占血液肿瘤的第二位。目前，MM的治疗取得了很大进展，患者的预后得到明显改善，但MM仍然是一种不可治愈的疾病。抗CD38单克隆抗体作为新的治疗药物被研究的越来越多，可以通过多种途径诱导MM细胞死亡。本文对抗CD38单克隆抗体在MM治疗中的临床应用作一综述。     

急性白血病患者SHP-1与JAK1 mRNA的表达与临床意义

目的探讨SHP-1和JAK1 mRNA在急性白血病（AL）患者的表达及其与AL复发及初次诱导缓解化疗疗效的关系。方法采用半定量反转录-聚合酶链反应（RT—PCR）的方法检测93例AL患者骨髓单个核细胞中SHP-1和JAK1 mRNA的表达,20例健康志愿者为健康对照。结果初治AL患者SHP-1 mRNA表达水平较健康对照组明显降低（P=0．000）,治疗完全缓解（CR）后表达增高（P=0．032）,复发时SHP-1 mRNA水平降低（P=0．015）;初治AL患者JAK1 mRNA表达水平较NC组略增高,但差异无统计学意义（P=0．051）,复发AL患者JAK1 mRNA表达水平较NC组增高,有统计学意义（P=0．047）;初治AL患者SHP-1 mRNA阳性组的诱导化疗CR率为88．89％,阴性患者组CR率为60．38％,差异有统计学意义（P=0．018）;SHP-1与JAK1 mRNA表达呈负相关（P=0．048）。结论AL患者中SHP-1 mRNA表达降低或缺如,SHP-1 mRNA阳性表达是初治AL患者预后良好的因素,并可作为判断疾病进展的预测指标。AL细胞中JAK1 mRNA丰度可能增高。     

原发性骨髓纤维化诊疗进展：第19届欧洲血液学会年会报道

原发性骨髓纤维化（PMF）属于慢性骨髓增殖性肿瘤的一种,JAK2V617F突变及其他分子学异常奠定了大多数费城染色体阴性的骨髓增殖性肿瘤的发病基础.从2005年第一次描述JAK2V617F突变到近期发现钙网蛋白（CALR）突变,再到JAK激酶抑制剂的临床应用,展示了对骨髓纤维化发病机制研究的深入和治疗手段的提高.现就2014年欧洲血液学会（EHA）年会上关于PMF的诊断、危险度分层、JAK激酶抑制剂治疗的益处和潜在毒副作用以及联合治疗的研究现状进行综述.     

丙泊酚抑制IL-6诱导的A549肺癌细胞上皮间质转化及其机制探讨

目的:探究丙泊酚对白细胞介素-6（IL-6）诱导的A549肺癌细胞上皮间质转化（EMT）的作用及机制。方法:将A549细胞随机分成四组:对照组、IL-6组（50 ng/ml重组IL-6蛋白）、IL-6+丙泊酚低剂量组（50 ng/ml重组IL-6蛋白和5 μmol/L丙泊酚）、IL-6+丙泊酚高剂量组（50 ng/ml重组IL-6蛋白和10 μmol/L丙泊酚）。MTT法检测细胞活力，Transwell检测细胞迁移情况，Real-time PCR方法检测EMT相关基因（E-cadherin、Vimentin和Snail1）mRNA的表达水平，Western blot检测EMT相关蛋白及JAK2和STAT3的磷酸化表达水平。使用0.5 μmol/L STAT3激活剂colivelin处理细胞，检测其对丙泊酚调节的IL-6诱导的A549细胞活力、迁移和EMT的影响。结果:与对照组相比，IL-6组中细胞的活力、迁移、EMT和JAK2/STAT3的活化均增加（P均<0.05）；与IL-6组相比，IL-6+丙泊酚组中细胞活力、迁移、EMT和JAK2/STAT3的活化均降低（P均<0.05），这些变化均具有剂量依赖性。STAT3激活剂能够减弱丙泊酚对IL-6诱导的A549细胞活力、迁移和EMT的影响（P均<0.05）。结论:丙泊酚能够抑制IL-6诱导的A549肺癌细胞EMT进程，这种作用是通过抑制JAK2/STAT3的活化发挥作用的。     

JAK2和STAT3在鼻咽癌中的表达及其与临床病理特征和预后的关系

目的:探讨JAK2和STAT3在鼻咽癌中的表达及其与临床病理特征和预后的关系。方法:收集2007年1月至2012年12月我院肿瘤科经病理证实的鼻咽癌患者103例,另取同期40例病理证实为慢性炎症的鼻咽黏膜标本作对照。免疫组化及Western Blot检测JAK2和STAT3在鼻咽癌组织以及鼻炎黏膜样本中的表达,并根据免疫组化结果统计其阳性率。进一步分析其与临床病理特征的关系;采用Spearman软件分析JAK2和STAT3在鼻咽癌中的相关性;采用Kaplan-Meier法检验JAK2和STAT3的表达与预后生存期的关系,并进行鼻咽癌预后的多因素分析。结果:免疫组化及Western Blot结果显示,鼻咽癌组织中JAK2和STAT3的表达明显高于鼻炎黏膜慢性炎组织（均P<0.05）。不同的T分期、N分期和临床分期中,JAK2和STAT3的阳性表达率差异具有统计学意义（均P<0.05）;不同年龄、性别、分化类型、吸烟及家族史之间,JAK2和STAT3的表达差异无统计学意义（均P>0.05）。经Spearman非参数检验的等级相关分析可知JAK2和STAT3的蛋白表达水平在鼻咽癌患者中存在正相关（均P<0.05）。Kaplan-Meier法结果提示JAK2和STAT3的表达水平与患者的生存期存在明显的负相关（均P<0.05）。鼻咽癌预后的多因素分析表明,T分期、N分期、临床分期以及JAK2和STAT3的表达是影响鼻咽癌预后的独立危险因素（均P<0.05）。结论:JAK2和STAT3在鼻咽癌组织中高表达,两者在鼻咽癌中的表达呈协同性,并与临床分期和淋巴结转移显著相关,JAK2和STAT3的高表达是鼻咽癌患者预后的独立危险因素。     

The role of JAK/STAT signalling in the pathogenesis,prognosis and treatment of solid tumours

Aberrant activation of intracellular signalling pathways confers malignant properties on cancer cells. Targeting intracellular signalling pathways has been a productive strategy for drug development, with several drugs acting on signalling pathways already in use and more continually being developed. The JAK/STAT signalling pathway provides an example of this paradigm in haematological malignancies, with the identification of JAK2 mutations in myeloproliferative neoplasms leading to the development of specific clinically effective JAK2 inhibitors, such as ruxolitinib. It is now clear that many solid tumours also show activation of JAK/STAT signalling. In this review, we focus on the role of JAK/STAT signalling in solid tumours, examining the molecular mechanisms that cause inappropriate pathway activation and their cellular consequences. We also discuss the degree to which activated JAK/STAT signalling contributes to oncogenesis. Studies showing the effect of activation of JAK/STAT signalling upon prognosis in several tumour types are summarised. Finally, we discuss the prospects for treating solid tumours using strategies targeting JAK/STAT signalling, including what can be learned from haematological malignancies and the extent to which results in solid tumours might be expected to differ.     

Stathmin 1 inhibition amplifies ruxolitinib-induced apoptosis in JAK2V617F cells

The JAK/STAT pathway is constitutively activated in myeloproliferative neoplasms and can be inhibited by ruxolitinib, a selective JAK1/2 inhibitor. The JAK2^V617F mutation leads to constitutive STAT3 phosphorylation and potentially leads to inhibition of Stathmin 1 activity via STAT3. In support of this hypothesis, we found that, in HEL JAK2^V617F cells, ruxolitinib treatment decreased STAT3 and Stathmin 1 association, induced Stathmin 1 activation and microtubule instability. Silencing of Stathmin 1 significantly reduced cell proliferation and clonal growth, and increased apoptosis induced by ruxolitinib. Stathmin 1 silencing also prevented ruxolitinib-induced microtubule instability. To phenocopy the effect of Stathmin 1 inhibition, cells were treated with paclitaxel, a microtubule-stabilizing drug, in association or not with ruxolitinib; combined treatment significantly increased apoptosis, when compared to monotherapy. Notably, Stathmin 1 mRNA levels were highly expressed in CD34⁺ cells from primary myelofibrosis patients. We then proposed that an undesired effect of ruxolitinib treatment may constitute Stathmin 1 activation and microtubule instability in JAK2^V617F cells. Induction of microtubule stability, through Stathmin 1 silencing or paclitaxel treatment, combined with ruxolitinib could be an effective strategy for promoting apoptosis in JAK2^V617F cells.     

Ruxolitinib induces apoptosis of human colorectal cancer cells by downregulating the JAK1/2-STAT1-Mcl-1 axis

Under pathological conditions, the Janus kinase (JAK)/STAT signaling pathway can regulate the proliferation, differentiation and migration of tumor cells, including colorectal cancer (CRC). CRC is the third major types of cancer among males and the second among females worldwide. In China, CRC is the fifth common cancer among both males and females. Western blotting, flow cytometry, RNA interference, immunoprecipitation, xenografts models, and immunohistochemical staining were carried out to evaluate the possible mechanisms of acton of ruxolitinib. The present data suggested that ruxolitinib can suppress CRC cell proliferation by inducing apoptosis. Firstly, JAK1/2-STAT1 was identified as the target of ruxolitinib. Then, ruxolitinib downregulated myeloid cell leukemia-1 (Mcl-1) mRNA level and decreased its protein level, which enabled Bak to trigger CRC apoptosis. Furthermore, ruxolitinib exerted potent activity against CRC xenograft growth in vivo. High expression of phosphorylated STAT1 (S727) was also confirmed in 44 pairs of human colon carcinoma and adjacent normal tissues. Taken together, the results showed that ruxolitinib decreased JAK1/2-STAT1-Mcl-1 protein level and effectively suppressed CRC cell proliferation in vitro and in vivo. Therefore, ruxolitinib could be a promising anticancer agent for CRC treatment.     

Genetic predisposition to leukemia and other hematologic malignancies

In this review, we provide an overview of familial myelodysplastic syndromes (MDS)/acute leukemia (AL) and bone marrow failure syndromes, as well as insights into familial myeloproliferative neoplasms (MPNs), familial multiple myeloma (MM), familial Waldenström macroglobulinemia (WM), familial lymphoma, and cancer predisposition syndromes with increased risk of MDS/AL. This field will continue to accelerate as next-generation sequencing (NGS) techniques identify novel predisposition alleles in families with a genetic predisposition to hematologic malignancies. Newly identified predisposition genes continue to inform the field of inherited leukemia and other hematologic malignancies. Current developments in clinical translation include techniques detailing the acquisition of appropriate germline material for patient work-ups, methods for genetic testing, and nuances essential for the treatment and clinical management of patients with a genetic predisposition to hematologic malignancies.     

c-FLIP mRNA 在恶性血液病中的表达及其意义

目的：探讨C—FLIPmRNA在恶性血液病中的表达及其意义。方法：用采用半定量逆转录聚合酶链反应（RT—PCR）检测42例恶性血液病骨髓单个核细胞C—FLIPmRNA的表达。包括急性白血病（AL）27例，其中初治21例及复发和完全缓解（CR）后AL各3例、慢性粒细胞性白血病（CML）5例、慢性中性粒细胞性白血病（CNL）1例和慢性淋巴细胞性白血病（CLL）4例，多发性骨髓瘤（MM）3例，骨髓增生异常综合征-难治性贫血伴原始细胞增多2型（MDS—RAEB-2）2例。结果：在初治和复发AL、初治CML、CNL、CLL、MDS—RAEB-2、MM中C—FLIPmRNA均呈异常增高表达，初治AL中c—FLIPmRNA的表达与复发AL比较差异无统计学意义（P〉0．05），其FAB各亚型之间的表达差异亦无统计学意义（P〉0．05）。初治AL与CLLC—FLIPmRNA的表达显著高于初治CML（P〈0．001），但初治AL与初治CLLE魄磋淠呒统计学意义（P〉0．05）。MDS—RAEB-2、MMC—FLIPmRNA的表达与AL的cFLIPmRNA表达均无统计学差异（P〉0．05）。对照组和CR后AL均为阴性表达。C—FLIPmRNA的表达与初治AL患者年龄、性别、初诊白细胞数、LDH以及核型、免疫表型无关。初治未达CR的AL患者其c—FLIPmRNA表达高于CR者，但并无统计学意义（尸〉0．05）。结论：恶性血液病C—FLIPmRNA的表达异常增高。C—FLIPmRNA能反映恶性血液病骨髓细胞的凋亡抑制情况，并与恶性血液病的类型、疾病状态、临床疗效和预后密切相关。     

Myeloproliferative and lymphoproliferative disorders: State of the art

Myeloproliferative neoplasms (MPNs), including polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF), are clonal disorders complicated mainly by vascular events and transformation to myelofibrosis (for PV and ET) or leukemia. Although secondary malignancies, in particular, lymphoproliferative disorders (LPNs), are rare, they occur at a higher frequency than found in the general population, and there has been recent scientific discussion regarding a hypothetical relationship between treatment with JAK inhibitors in MPN and the risk of development of LPN. This has prompted increased interest regarding the coexistence of MPN and LPN. This review focuses on the role of JAK2 and the JAK/STAT pathway in MPN and LPN, whether there is a role for the genetic background in the occurrence of both MPN and LPN and whether there is a role for cytoreductive drugs in the occurrence of both MPN and LPN. Furthermore, whether an increased risk of lymphoma development is limited to patients who receive the JAK inhibitor ruxolitinib, is a more general phenomenon that occurs following JAK1/2 inhibition or is associated with preferential JAK1 or JAK2 targeting is discussed.