非小细胞肺癌患者外周血和肺癌组织中ZIC1启动子甲基化检测的临床意义

目的 探讨ZIC1基因在NSCLC患者外周血和肺肿瘤组织中的甲基化状态,及其对NSCLC的预后评估意义.方法 选取95例NSCLC患者外周血、癌和癌旁组织,另选取95例健康体检者外周血为对照组.用MSP法比较外周血和癌组织的ZIC1甲基化检出率;分析ZIC1在外周血和癌组织的甲基化和NSCLC临床病理因素的相关性.结果...     

ERCC1在恶性肿瘤患者外周血和肿瘤组织中表达的相关性研究

目的 探讨核苷酸切除修复交叉互补基因1（ERCC1）在肺癌、结肠癌、胃癌、乳腺癌患者外周血和肿瘤组织中表达的相关性及其临床意义。方法 采用实时荧光定量PCR（RT-qPCR）技术检测230例肺癌、160例结肠癌、125例胃癌、100例乳腺癌患者的肿瘤石蜡标本及其对应的血液样本中ERCC1 mRNA的表达水平。结果 ERCC1 mRNA在4种肿瘤的癌组织和外周血中的表达差异均无统计学意义（P＞0.05）。4种肿瘤的癌组织中ERCC1 mRNA的相对表达量高于相对应外周血中的表达（P＜0.05）,且肿瘤组织与外周血ERCC1 mRNA的表达均呈正相关（肺癌：r=0.430,P＜0.001;结肠癌：r=0.553,P＜0.001;胃癌：r=0.583,P＜0.001;乳腺癌：r=0.501,P＜0.001）。肺癌、结肠癌及乳腺癌肿瘤组织和外周血中ERCC1的表达水平与年龄、性别均无关（P＞0.05）。胃癌肿瘤组织和外周血中ERCC1 mRNA的表达水平与年龄无关,而男性患者肿瘤组织中ERCC1的表达高于女性（P＜0.05）。结论 通过检测肺癌、结肠癌、胃癌及乳腺癌患者外周血中ERCC1的表达水平能够在一定程度上间接反映癌组织中ERCC1的表达状况。     

胃癌患者外周血与胃癌组织中ERCC1基因甲基化的关系及其意义

背景与目的：胃癌的发生基于基因和表观遗传学机制,表观遗传学的改变在胃癌的发展中起到重要作用。DNA甲基化是目前研究最多、最为深入的一种表观遗传学表达机制。DNA甲基化是一个可逆性过程。核苷酸切除修复交叉互补基因1(excision repair cross-complementing gene 1,ERCC1)是一种DNA损伤修复基因。本研究检测胃癌患者外周血与胃癌组织中ERCC1基因启动子CpG岛甲基化状态,探讨两者的关系及其意义。方法：采用甲基化特异性PCR技术,检测30例胃癌患者外周血、胃癌组织中ERCC1基因启动子CpG岛甲基化状态。结果：胃癌组织中ERCC1基因启动子CpG岛甲基化率为76.7%(23/30),外周血中ERCC1基因启动子CpG岛甲基化率为63.3%(19/30),差异无统计学意义。结论：胃癌患者外周血中的ERCC1基因启动子CpG岛甲基化率与胃癌组织中相似,检测胃癌患者外周血中的ERCC1基因启动子CpG岛甲基化状态为治疗胃癌提供一个简便、快捷、可靠的途径,同时也为以ERCC1基因启动子CpG岛甲基化作为靶点治疗胃癌提供了可靠的理论依据。     

实时荧光定量RT-PCR检测肺癌、癌旁组织及外周血中CK19 mRNA的表达及临床意义

目的 探讨角蛋白CK19在肺癌患者癌和癌旁组织及外周血循环细胞中的表达特点及其临床意义。方法 采用实时荧光定量RT-PCR的方法对62例肺癌患者癌和癌旁组织内CK19mRNA表达进行检测，并与其对应的外周血中的CK19 mRNA表达结果进行比较。结果 62例肺癌患者的癌和癌旁组织和外周血中CK19mRNA均有表达（分别为61．2％、32．2％、40．3％），而肺癌组织中CK19mRNA水平明显高于癌旁组织，两者差异显著（P〈0．05）。外周血中CK19mRNA的表达与临床分期和病理类型无关；却与肺癌组织中CK19mRNA的高表达水平及淋巴结是否转移密切相关；健康对照组则无阳性。结论 实时荧光定量RT-PCR方法检测肺癌患者外周血中CK19 mRNA表达有较高的敏感性和特异性，且操作方便，在肿瘤微转移的诊断监测方面有一定的应用价值。     

大肠癌有丝分裂关卡基因ＢＵＢ１和ＢＵＢＲ１ ｍＲＮＡ的表达分析

目的　研究大肠癌组织中有丝分裂关卡基因ＢＵＢ１和ＢＵＢＲ１ｍＲＮＡ的表达。方法　采用半定量ＲＴ-ＰＣＲ方法检测３６例大肠癌和癌旁正常组织中ＢＵＢ１和ＢＵＢＲ１ｍＲＮＡ表达水平。结果　大肠癌和癌旁正常组织中的ＢＵＢ１ｍＲＮＡ拷贝数／β-ａｃｔｉｎｍＲＮＡ拷贝数比值分别为０.６７±０.３３和１.２４±０.３７;大肠癌和癌旁正常组织中的ＢＵＢＲ１ｍＲＮＡ拷贝数／β-ａｃｔｉｎｍＲＮＡ拷贝数比值分别为０.５３±０.２５和１.０３±０.４８。ＢＵＢ１和ＢＵＢＲ１ｍＲＮＡ的表达水平在大肠癌中显著低于癌旁正常组织（Ｐ＜０.０５）。结论　ＢＵＢ１和ＢＵＢＲ１基因是大肠癌的肿瘤相关基因,可能在大肠癌发病中具有重要作用。     

外周血人类斯钙素-1基因表达与乳腺癌微转移的关系

目的:检测乳腺癌患者外周血、肿瘤组织中hSTC-1mRNA的表达,判断外周血微转移状况,探讨微转移与肿瘤恶性程度之间的关系。方法:采用RT-PCR法检测乳腺癌组织、外周血中hSTC-1的mRNA表达,用免疫组化法检测乳腺肿瘤组织中ER、PR及VEGF的表达。结果:hSTC-1mRNA在乳腺癌患者癌组织中的阳性率为93.3%(28/30),在外周血中的阳性率为37.3%(19/51);17例非肿瘤成年女性患者外周血中未检测到hSTC-1的mRNA。血液中的hSTC-1的mRNA表达与多个临床病理因子有相关性(P<0.05),包括:肿瘤大小、腋淋巴结转移、TNM分期、ER、VEGF等;结论:乳腺癌患者外周血检测到hSTC-1基因的表达,说明乳腺癌已发生血行微转移,乳腺癌患者外周血hSTC-1的mRNA表达与多个临床病理因子有关;这些因子决定乳腺癌的预后,因此hSTC-1可以作为检测乳腺癌外周血微转移的一种新的分子标记。     

食管癌外周血及肿瘤组织的EGFR突变的检测及意义

目的：分析食管癌患者外周血及肿瘤组织EGFR突变与临床特征关系,探讨外周血及肿瘤组织中EG-FR突变的一致性,进一步分析EGFR-TKI治疗外周血和肿瘤组织中EGFR突变与临床疾病控制率之间的关系,进一步探讨EGFR突变与吉非替尼或者厄洛替尼临床疗效监测中的应用价值。方法采用RT-PCR检测112例食管癌患者外周血和87例食管癌患者肿瘤组织中EGFR突变,其中47例外周血和肿瘤组织配对检测。分析外周血和肿瘤患者组织中EGFR突变与临床特征的关系及两者EGFR突变的一致性。采用EGFR-TKI治疗外周血和肿瘤组织中EGFR突变,评价EGFR突变与吉非替尼或者厄洛替尼临床疗效的关系。结果 EGFR突变在鳞癌组明显高于非鳞癌组（P﹤0.05）;肿瘤组织和外周血同时EGFR突变的有11例,且突变位点一致,检测到的EGFR状态以肿瘤组织为标准,两者突变一致性达到73.33%;外周血和肿瘤组织中EGFR突变的患者接受TKI的疾病控制率明显高于EGFR野生型患者（P﹤0.01）。结论食管癌外周血和肿瘤组织EGFR突变与临床特征的关系与鳞癌密切相关;食管癌外周血和肿瘤组织EGFR突变的一致性较高;EGFR-TKI分子靶向治疗外周血和肿瘤组织中EGFR突变的疗效显著。     

PD-1/PD-L1抑制剂与多药联合治疗三阴性乳腺癌的研究进展

因缺乏明确的分子靶点,三阴性乳腺癌（TNBC）与其他乳腺癌亚型相比是较难治疗以及预后较差的乳腺癌亚型。程序性死亡受体-配体1（PD-L1）在TNBC中表达明显增加,其免疫逃逸机制使其更适用于免疫检查点阻断治疗。然而,临床试验中TNBC对抗PD-L1或抗程序性死亡受体1（PD-1）治疗的反应率并不令人满意,客观反应率只有10%~20％,可能与TNBC具有免疫沉默效应有关,因此免疫检查点阻断剂与其他药物的联合策略可能会增加TNBC的临床获益率。本文综述TNBC中PD-1/PD-L1抑制剂联合其他药物的研究进展,为临床医生设计多药联合策略提供依据。     

卵巢癌组织中SSBP1的表达意义及作用

目的 探讨线粒体DNA单链结合蛋白SSBP1在卵巢癌(Ovarian cancer)组织中的表达水平变化及其意义.方法 从The Cancer Genome Atlas(TCGA)数据库中选取经转录组测序分析的正常卵巢组织97例及卵巢癌组织373例,分析癌组织中SSBP1的表达变化,并利用Kaplan-Meier模型分...     

PD-L1与PD-1在宫颈鳞癌与宫颈良性病变组织及外周血中的表达及其预后价值

目的:通过分别对宫颈鳞癌、宫颈良性疾病组织和配对血液标本检测,了解外周血程序性死亡受体1(PD-1)与肿瘤组织程序性死亡受体-配体1(PD-L1)表达的一致性,初步评价其与宫颈鳞癌预后的关系。方法:收集宫颈鳞癌及宫颈良性病变组织标本各70例,并收集配对的外周血标本鳞癌24例,良性病变30例。免疫组化法检测组织PD-L1,流式细胞术检测外周血T细胞表面PD-1+亚群的比例,并结合生存资料做统计分析。结果:宫颈鳞癌患者外周血PD-1与组织PD-L1表达呈正相关（r2=0.734,P=0.000;r2=0.66,P=0.000）,宫颈良性病变者外周与组织的表达不相关（r2=0.138,P=0.043;r2=0.174,P=0.022）。宫颈癌患者血CD8+PD-1+细胞百分比、组织PD-L1+的细胞数大于宫颈良性病变组（P=0.000,P=0.002）。宫颈癌患者随肿瘤直径增大、分期升高,肿瘤组织PD-L1表达量也相应升高,外周T细胞表面PD-1表达也有类似趋势。但是外周T细胞表面PD-1表达程度与肿瘤大小、分化程度、深肌层侵犯均无关。单因素分析提示外周CD4+T细胞低表达PD-1的、组织低表达PD-L1的患者生存期较长。多因素分析结果显示,PD-L1表达对患者生存有影响（B=1.844,P=0.019）。结论:宫颈鳞癌患者外周血PD-1、组织PD-L1表达与肿瘤的分级、预后有关,组织PD-L1表达是患者的独立预后因素。     

Immunological profile of patients with ovarian-cancer under immunostimulation with murine monoclonal-antibodies

The longer survival of ovarian cancer patients after immunostimulation has been connected with the induction of an anti-tumor activity triggered by cellular and humoral immune responses. Our interest was to study the long-term influence on the immune system in relation to the various levels of the HAMA response and the disease stage. The immunological profile was evaluated by regularly performing phenotyping and functional analysis of peripheral blood lymphocytes (PBL). We report the statistical analysis of immunological data obtained in a study with 77 ovarian cancer patients examined over a period of up to 28 months. The results demonstrate that these immunological data are important for monitoring cancer patients under immunotherapy whereas they provide no prognostic significance.     

Skp1蛋白在非小细胞肺癌组织和外周血中的表达水平及其临床意义

目的：检测S期激酶相关蛋白1（Skp1）在非小细胞肺癌（NSCLC）组织和外周血中的表达水平并探讨其临床意义。方法：采用Western blot检测34例NSCLC组织及相应癌旁组织中Skp1蛋白的表达水平。采用组织芯片和免疫组织化学染色检测Skp1在86例NSCLC组织及相应癌旁组织中的表达情况,并分析Skp1蛋白的表达与NSCLC临床病理指标及患者预后的关系。采用酶联免疫吸附试验（ELISA）检测39例NSCLC患者及27例健康人血浆中的Skp1蛋白水平。结果：Western blot和免疫组化法分析结果显示Skp1蛋白在NSCLC组织中的表达水平均显著高于相应癌旁组织（P < 0.001）。ELISA法分析结果显示NSCLC病人血浆中的Skp1蛋白表达水平显著高于健康人群（P=0.003）。Kaplan-Meier单因素生存分析显示Skp1蛋白在NSCLC组织中的高表达与中晚期（Ⅱ+Ⅲ） NSCLC患者的不良预后显著相关（P=0.001）,多因素Cox回归分析显示Skp1蛋白能够作为影响中晚期（Ⅱ+Ⅲ） NSCLC患者预后的独立危险因素。Skp1蛋白表达水平与病理类型、临床分期、分化程度、淋巴结转移等因素无显著相关关系（P > 0.05）。结论：Skp1蛋白在NSCLC患者中高表达,并且Skp1蛋白在NSCLC组织中的高表达与中晚期（Ⅱ+Ⅲ） NSCLC患者的不良预后显著相关,可作为中晚期NSCLC患者预后的独立危险因素。     

散发性乳腺癌患者血浆BRCA1基因启动子异常甲基化的检测

目的:探讨散发性乳腺癌组织及外周血浆中BRCA1基因启动子异常甲基化状况及其在散发性乳腺癌诊断中的价值。方法:用甲基化特异PCR方法对散发性乳腺癌患者癌组织、癌旁组织及相应血浆进行BRCA1异常甲基化检测。结果:93例散发性乳腺癌组织中,BRCA1基因启动子异常甲基化率为29%(27/93),相应血浆中BRCA1的甲基化检出率为24.7%(23/93),而癌旁组织、正常对照血浆未检出甲基化,只检出未甲基化的BRCA1。血浆中甲基化改变与肿瘤组织甲基化状况显著相关(P<0.05);BRCA1异常甲基化与髓样癌和粘液腺癌的组织分型显著相关(P<0.05),也与肿瘤淋巴结转移显著相关(P<0.005);但与散发性乳腺癌患者年龄(绝经与否)及肿瘤分级无显著的相关性(P>0.05)。结论:血浆BRCA1基因异常甲基化改变的检测在散发性乳腺癌的特异诊断、组织分型和淋巴结恶性转移等方面有一定的应用价值。     

Perioperative biobehavioral interventions to prevent cancer recurrence through combined inhibition of β-adrenergic and cyclooxygenase 2 signaling

Evidence suggests that excess perioperative activation of the sympathetic nervous system and the consequent release of catecholamines (ie, epinephrine and norepinephrine) in the context of cancer surgery and inflammation may significantly facilitate prometastatic processes. This review first presents biomedical processes that make the perioperative timeframe pivotal in determining long-term cancer outcomes nonproportionally to its short duration (days to weeks). Then, it analyzes the various mechanisms via which the excess release of catecholamines can facilitate the progression of cancer metastases in this context by directly affecting the malignant tissues and by regulating, via indirect pathways, immunological and other mechanisms that affect metastatic progression in the tumor microenvironment and systemically. In addition, this review addresses the need to supplement β-adrenoreceptor blockade with cyclooxygenase 2 inhibition, especially during surgery and shortly thereafter, because similar mechanisms are simultaneously activated by surgery-induced inflammatory responses. Importantly, this review presents translational and clinical evidence showing that perioperative β-adrenoreceptor blockade and cyclooxygenase 2 inhibition can reduce the prometastatic process and cancer recurrence, and the clinical feasibility and safety of this approach are demonstrated as well. Lastly, alternative psychophysiological approaches to the use of β-adrenergic blockers are presented because a substantial portion of patients have medical contraindications to this pharmacological treatment. The adaptation of existing psychophysiological interventions to the perioperative period and principles for constructing new approaches are discussed and exemplified. Overall, pharmacobehavioral interventions, separately or in combination, could transform the perioperative timeframe from being a prominent facilitator of metastatic progression to an opportunity for arresting or eliminating residual disease, potentially improving long-term survival rates in cancer patients.     

Semaphorin 4C promotes motility and immunosuppressive activity of cancer cells via CRMP3 and PD-L1

Semaphorins (SEMAs) are membrane-bound or soluble proteins that participate in organ development and cancer progression, however, the detailed role of SEMAs in carcinogenesis is not fully elucidated yet. Our in silico analysis showed among the differentially expressed SEMAs in colon cancer tissues, patients with higher SEMA4C expression tumors had worse survival. The migration and invasion of the HCT116 and CT26 colon cancer cells were significantly suppressed by SEMA4C neutralizing antibody treatment; while enhanced by ectopic expression of SEMA4C. Subsequently, RNA sequencing study revealed microtubule polymerization- and nucleation-related genes are highly enriched in SEMA4C overexpression HCT116 cells. Western blotting showed the negative correlation between the levels of SEMA4C expression and tubulin acetylation. Mechanistic study showed SEMA4C interacted with and stabilized collapsin response mediator protein 3 (CRMP3), a novel deacetylase, to increase α-tubulin deacetylation and cell motility, which could be effectively attenuated after HDAC inhibitors treatment. We also found that a tumor-suppressive miRNA let-7b can target SEMA4C and act synergistically with SEMA4C neutralizing antibody to suppress the motility of colon cancer cells. In addition, blockade of SEMA4C could attenuate the expression of program death ligand 1 (PD-L1). Collectively, our results highlight that SEMA4C may promote colon cancer progression through modulating CRMP3-mediated tubulin deacetylation and PD-L1-mediated immunosuppression.     

SRC1在胃癌中的表达及其与预后的关系

目的 探讨类固醇受体辅助活化因子-1(Steroid receptor coactivator 1,SRC1)蛋白在胃癌组织中的表达及其与患者临床病理参数和预后的相关性。方法 收集36例胃癌组织及其配对的癌旁组织,qRT-PCR法及Western blot法检测SRC1mRNA和蛋白的表达水平。应用免疫组化方法检测286例胃癌组织中SRC1蛋白的表达情况,分析SRC1蛋白表达与胃癌临床病理参数的关系及其对患者预后的影响。结果 与癌旁组织相比,胃癌组织中SRC1mRNA表达水平明显降低(P=0.004),SRC1蛋白表达水平也明显降低。SRC1蛋白的表达强度与胃癌患者临床病理参数无显著关系。Kaplan-Meier生存曲线分析发现高表达SRC1组5年生存率显著高于低表达组(P=0.009)。Cox回归分析结果显示低表达SRC1(P=0.002)、肿瘤侵袭T_4a~T_4b(P=0.004)、淋巴结转移N(P=0.038)、远处转移M1(P＜0.001)是独立影响总生存时间的预后不良因素。SRC1低表达较高表达患者预后差。结论 胃癌中SRC1的表达明显降低,胃癌组织中的SRC1表达水平可作为独立的胃癌预后不良因素,其过低表达与胃癌的发展密切相关,可能作为一个抑癌基因及判断胃癌患者预后的标志物。     

Distinct chemotherapy‐associated anti‐cancer immunity by myeloid cells inhibition in murine pancreatic cancer models

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy associated with an extremely poor prognosis. Chemotherapy, such as gemcitabine (GEM), is the only treatment for PDAC patients who are not suitable for radical surgical treatment; however, its anti‐tumor efficacy is limited. In this study, we investigated the host immune system response in murine PDAC models undergoing GEM treatment. We found that PDAC tumor tissues were infiltrated with a substantial number of Gr‐1+ myeloid cells and had relatively small numbers of CD4+ and CD8+ cells. In addition, there were increased numbers of myeloid cells expressing CD11b+ and Gr‐1+ in peripheral blood. When mice with PDAC tumors in the intraperitoneal cavity or liver were treated with GEM, numbers of myeloid cells in tumor tissues and in peripheral blood decreased. In contrast, numbers of CD4+ or CD8+ cells increased. In peripheral blood, the numbers of CD8+ cells expressing interferon‐gamma (IFN‐γ) were higher in GEM‐treated mice than in untreated mice. In addition, GEM treatment in combination with myeloid cell depletion further prolonged the survival of PDAC mice. The gene expression profile of peripheral blood in myeloid cell‐depleted PDAC mice treated with GEM showed biological processes related to anti‐cancer immunity, such as natural killer cell‐mediated cytotoxicity, type I IFN signaling, and co‐stimulatory signaling for T cell activation. Thus, in PDAC murine models, GEM treatment was associated with an immune response consistent with an anti‐cancer effect, and depletion of myeloid‐lineage cells played an important role in enhancing anti‐cancer immunity associated with GEM treatment.     

大肠癌患者外周血OD4＋0D25＋CD127－调节性T细胞表达及其临床意义

目的：探讨OD4＋0D25＋CD127－调节性T细胞在大肠癌患者外周血中的表达水平及临床意义。方法：应用流式细胞仪检测200例大肠癌患者外周血OD4＋0D25＋CD127－调节性T细胞占CD4＋T细胞的百分比,并分析其与大肠癌组织的分化程度、淋巴结转移和临床分期的关系。结果：大肠癌患者外周血OD4＋0D25＋CD127－调节性T细胞占CD4＋T细胞的百分率[（4．84±1．35）％]明显高于健康对照组[（0．85±0．25）％],差异有统计学意义,P〈0．05。低分化者外周血调节性T细胞[（4．21±0．42）％]明显高于高分化者[（3．92±0．41）％],差异有统计学意义,P〈0．05;有淋巴结转移者外周血调节T细胞[（4．57±l_44）％]明显高于无淋巴结转移者[（2．36±0．68）％],差异有统计学意义,P〈0．01;Ⅲ和Ⅳ期患者外周血调节性T细胞[（3．53±1．41）％和（4．38±1．32）％]明显高于Ⅰ～Ⅱ期[（1．90±0．86）％],差异有统计学意义,P〈0．01。结论：大肠癌患者外周血OD4＋0D25＋CD127调节性T细胞占CD4＋T细胞的百分率明显升高,可能在大肠癌的免疫耐受和免疫逃逸中发挥重要作用,检测其结果对于判断大肠癌的病程进展及预后有一定参考价值。