造血干细胞移植支持下大剂量化疗对高危乳腺癌患者血清细胞因子的影响

目的:观察造血干细胞移植支持下的大剂量化疗对高危乳腺癌患者Th1、Th2类细胞因子(IFN-γ、IL-2,IL-10)的影响。方法:应用ELISA方法测定14例高危乳腺癌患者大剂量化疗前后血清IFN-γ、IL-2、IL-10的变化。结果:14例患者大剂量化疗前后血清中IFN-γ、IL-2、IL-10的水平分别为12.79±4.60pg/ml、6.55±5.42pg/ml、9.04±8.28pg/ml及12.38±3.23pg/ml、6.16±2.84pg/ml、9.00±6.85pg/ml,前后对比无统计学意义(P>0.05)。不同病理类型患者大剂量化疗前后血清细胞因子亦无显著差异(P>0.05)。转移淋巴结数大于50%与少于50%组相比,IL-10水平在大剂量化疗前两组有显著差异(P<0.05),但化疗后这种差异消失(P>0.05)。结论:大剂量化疗对Th1及Th2类细胞因子无影响,而对淋巴结转移数大于50%者的Th2类细胞因子有调节作用。     

干细胞基因Lgr5在食管鳞癌组织中的表达及其临床意义

目的 探讨干细胞基因Lgr5在食管鳞癌组织及其成球细胞中的表达意义。方法 采用免疫组织化学法（IHC）检测280例食管鳞癌和150例癌旁组织中Lgr5蛋白的表达水平;低黏附培养法培养食管鳞癌细胞KYSE70形成成球细胞,Quantitative real-time PCR（qRT-PCR）、Western blot法检测干性相关基因SOX2、ALDH1A1、NANOG以及Lgr5在亲代细胞和成球细胞中的表达差异;Transwell实验检测成球细胞的迁移和侵袭能力、qRT-PCR法检测成球细胞中上皮间质转化（epithelial-mesenchymal transition, EMT）相关标志物Vimentin和N-cadherin的表达水平。结果 Lgr5在食管鳞癌组织的表达水平明显高于癌旁正常食管组织（P＜0.05）,且与淋巴结转移、浸润深度、肿块大小、分化程度、肿瘤分期密切相关（P均＜0.05）。与亲代细胞相比,食管鳞癌成球细胞中干性相关基因SOX2、ALDH1A1、NANOG表达水平升高（P均＜0.05）,Lgr5在食管鳞癌成球细胞中的表达水平也明显升高（P＜0.05）,并且其迁移和侵袭能力增强（P＜0.05）,EMT相关标志物Vimentin和N-cadherin的mRNA表达水平升高（P均＜0.05）。结论 Lgr5可能为食管鳞癌复发、进展、侵袭转移的标志之一,Lgr5高表达的食管鳞癌细胞可能具有干细胞样特性,并且可能为食管鳞癌细胞的一个干性调控基因。     

Cortactin在非小细胞肺癌中的表达及其与临床特征的关系探讨简

目的：探讨cortactin在非小细胞肺癌中表达与临床特征的相关性.方法：回顾性分析77例非小细胞肺癌患者的临床病历资料.应用免疫组织化学技术,检测非小细胞肺癌癌组织及正常组织的cortactin表达情况,并进行分析.结果：Cortactin 在正常肺组织的血管旁平滑肌细胞胞浆中阳性表达,其阳性率为22.1% （17/77）.在NSCLC 组织中,Cortactin 于癌细胞胞浆阳性表达,其阳性率为58.4%（45/77）,与正常肺组织之间比较具有显著性差异（P＜0.05）.Cortactin阳性表达与肿瘤分化程度呈负相关,与TNM分期、淋巴结转移呈正相关.Cortactin阳性表达与患者性别、是否吸烟、病理类型均无明显关系.结论：Cortactin阳性表达高低与NSCLC的临床分期、分化程度及有无淋巴结转移有一定关系,可以作为NSCLC预后的一个参考指标.     

Lgr5表达及Kras突变与结直肠癌临床病理特征的相关性研究

目的 探讨Lg6表达及Kras基因突变与结直肠癌临床病理特征的关系.方法 采用免疫组织化学方法,检测539例结直肠癌患者癌组织中Lgr5的表达,应用聚合酶连反应(polymerase chain reaction,PCR)扩增DNA直接测序法检测Kras基因第12、13密码子突变,并分析其与临床病理特征的关系.结果 在539例结直肠癌中,Lgr5总体表达率为66.79％ (360/539),在不同组织学类型、分化程度及病理分期分组中,其表达差异有统计学意义(P＜0.05),与肿瘤分化(γ =0.102,P=0.018)、肿瘤分期(γ=0.093,P=0.030)、浸润深度(γ=0.131,P=0.002)及是否有脉管和神经的侵犯(γ=0.087,P =0.044)呈正相关性;Kras基因突变总突变检出率为37.29％ (201/539),Kras基因突变与性别及结肠部位有关(P＜0.05),女性及右半结肠的突变率较高.Lgr5表达与Kras基因突变间无相关性.结论 Lgr5表达与肿瘤的发生发展相关,提示Lgr5可能是治疗的潜在靶点.Kras基因突变在女性及右半结肠癌中检出率较高,突变类型与Lgr5表达无显著相关性.     

原发性乳腺癌组织中雄激素受体的表达及其与临床病理指标的关系

目的探讨雄激素受体（AR）在原发性乳腺癌组织中的表达情况及其与乳腺癌临床病理指标的关系。方法采用免疫组织化学方法检测520例原发性乳腺癌患者肿瘤组织中AR、ER、PR、HER-2、Ki-67的表达情况,并结合患者年龄、月经状态,淋巴结转移情况、肿瘤大小、病理类型、TNM分期以及肿瘤组织学分级等临床病理指标进行分析。结果乳腺癌组织中AR的阳性率为81.3%,肿瘤越小者AR的阳性表达率越高（P=0.000）,组织学分级越低者AR的阳性表达率越高（P=0.029）,ER、PR阳性表达越强者AR的阳性率越高（P=0.000,P=0.000）。AR的表达与年龄、淋巴结转移、病理类型、脉管瘤栓、TNM分期、HER-2及Ki-67等无关。结论AR在乳腺癌组织中广泛表达,是乳腺癌恶性程度低、预后良好的指标;对AR阳性患者有望通过针对AR的途径来给予治疗。     

乳腺黏液腺癌的超声表现及其与病理特征的关系

目的探讨乳腺黏液腺癌超声表现与病理特征的关系。方法于彩超下观察乳腺黏液腺癌患者的超声表现,根据其病理特征分为单纯型、混合型2组,分析超声表现与病理特征的关系。结果混合型乳腺黏液腺癌患者中检测到动脉血流频谱占88.6%,高于单纯型乳腺黏液腺癌患者,差异有统计学意义(P<0.05);2组患者的峰值血流速度、阻力指数无明显差异(P>0.05);混合型乳腺黏液腺癌患者血流信号达2级、3级者分别为37.1%、20.0%,明显高于单纯型乳腺黏液腺癌患者,差异有统计学意义(P<0.05)。2组患者的肿瘤边界、肿瘤边缘、肿瘤后方回声分型、瘤内钙化有显著差异(P<0.05);在回声类型、肿瘤形态无明显差异(P>0.05)。结论与单纯型乳腺黏液腺癌相比,混合型黏液腺癌在超声下具有动脉血流频谱、血流信号分级升高,肿瘤边界不清、边缘模糊、肿瘤后方回声增强、瘤内出现钙化的特点。     

乳腺癌干细胞上皮-间质转化标志物表达变化及意义

目的 探讨乳腺癌干细胞上皮．间质转化标志物表达变化及其临床意义。方法采用无血清悬浮培养法,从MCF-7细胞培养乳腺癌微球体细胞,应用流式细胞仪检测微球体细胞中CD44和CD24的表达,采用Westernblot方法检测微球体细胞中E-钙黏素、N-钙黏素、纤维连接蛋白、波形蛋白的表达水平,体外穿膜实验检测肿瘤细胞的迁移、侵袭能力。结果乳腺微球体富集了CD44＋CD24-的乳腺癌干细胞,并能在含血清培养基中增殖分化。该微球体细胞的上皮标志物E-钙黏素表达水平下调,而间质标志物N-钙黏素、纤维连接蛋白、波形蛋白的表达水平上调,同时乳腺癌干细胞的迁移、侵袭能力显著增强。结论乳腺癌干细胞具有上皮-间质转化的特征,具有显著增强的迁移、侵袭能力。     

乳腺癌PCNA表达与肥大细胞密度的关系

 目的　探讨乳腺癌组织中PCNA表达与MC密度的关系。方法　用免疫组织化学方法和俾士麦棕法分别检测100例乳腺癌组织中增殖细胞核抗原(PCNA)和肥大细胞(MC)。结果　1.PCNA阳性表达与乳腺癌分化程度和淋巴结转移相关,高分化癌阳性表达低于中分化癌和未分化癌,P<0.05;淋巴结有转移者PCNA阳性表达高于无转移者,P<0.01。2. PCNA阳性者,MC计数较低;PCNA阴性者,MC计数较高,乳腺癌PCNA表达与癌周MC密度呈负相关。结论　PCNA是判断乳腺癌预后的有效指标,MC可能通过影响PCNA的表达发挥其抗肿瘤作用。     

Clinicopathological Significance of CD133 and ALDH1 Cancer Stem Cell Marker Expression in Invasive Ductal Breast Carcinoma

Background: Biomarkers in breast neoplasms provide invaluable information regarding prognosis and help determining the optimal treatment. We investigated the possible correlation between cancer stem cell (CSC) markers (CD133, and ALDH1) in invasive ductal breast carcinomas with some clinicopathological parameters. Aim: To assess the correlation between expression of cancer stem cell (CSC) markers (CD133, and ALDH1) and clinicopathological parameters of invasive ductal breast carcinomas. Materials and Methods: Immunohistochemical analysis of CD133 and ALDH1 was performed on a series of 120 modified radical mastectomy (MRM) specimens diagnosed as invasive ductal breast carcinoma. Results: Expression of both CD133 and ALDH1 was significantly changed and related to tumor size, tumor stage (TNM), and lymph node metastasis. A negative correlation between CD133 and ALDH1 was found. Conclusions: Detecting the expression of CD133 and ALDH1 in invasive ductal breast carcinomas may be of help in more accurately predicting the aggressive properties and determining the optimal treatment.     

肺癌组织中 HMGN5基因的表达水平及与其临床病理特征的相关性分析

目的：探讨高迁移率族核小体结合域5（ HMGN5）基因在肺癌组织中的表达水平及与其临床病理特征的相关性分析。方法选择手术切除的肺癌组织标本共75例,同期取20例肺癌患者癌旁正常组织作为对照组;应用RT-PCR法检测HMGN5基因在肺癌组织中的表达;分析HMGN5基因的表达水平与肺癌组织临床病理特征的关系。结果肺癌组HMGN5基因转录mRNA水平明显高于癌旁组,比较差异有统计学意义（ P＜0．01）;肺癌组织中HMGN5基因mRNA水平与患者年龄、性别、病理类型无相关性（P＞0．05）,而与病理分级、淋巴结转移以及临床分期有明显相关性（P＜0．01）。结论肺癌组织中HMGN5基因mRNA呈高水平表达,并与病理分级、淋巴结转移以及临床分期呈显著有关;提示HMGN5基因的表达水平可能与肺癌的发生、发展密切相关。     

DNA—PKcs在乳腺癌组织中的表达与分期相关性研究

目的：探讨乳腺癌组织中DNA依赖蛋白激酶（DNA dependent protein kinase，DNA—PK）的催化亚单位DNA—PKcs的表达变化与分期的相关性。方法：采用免疫组化SP法检测30例Ⅰ期乳腺癌组织，11例ⅢB、ⅢC期乳腺癌组织及19例Ⅳ期乳腺癌组织中DNA-PKcs的表达情况。结果：DNA-PKcs在Ⅰ期乳腺癌组织中和其癌旁正常组织中均呈高表达，两者表达具有一致性（P〉0．05）。DNA-PKcs在ⅢB、ⅢC期和Ⅳ期乳腺癌组织中的表达明显低于Ⅰ期乳腺癌（P〈0．05和P〈0．01）。同时，把ⅢB、ⅢC期和Ⅳ期合并后，与Ⅰ期比较，DNA—PKcs表达差异有极显著性意义（P〈0．01）。结论：DNA．PKcs的低表达在乳腺癌发生进展和远处转移中有重要意义，对乳腺癌的分期有一定指导意义，可能成为乳腺癌分期的生物学指标。     

超声显像特征与乳腺癌组织中HER-2表达的关系研究

目的分析乳腺癌中超声显像特征与乳腺癌组织HER-2表达的关系。方法回顾性分析女性乳腺癌患者共84例,收集其术前检查的超声显像特征,并收集其病理结果中免疫组化法检测的HER-2表达的结果,分析超声显像特征与乳腺癌组织HER-2表达的关系。结果 84例女性乳腺癌患者中,其中HER-2表达为阴性的共21例,弱阳性共26例,阳性29例,强阳性的8例,其中低表达56. 0%,高表达44%,超声特征中乳腺癌的HER-2表达水平与肿瘤大小、内部钙化呈显著相关性(P <0. 05),而乳腺癌的HER-2表达水平与边缘情况、肿瘤形状、肿瘤纵横比、内部回声、后方回声衰减和血供分级没有显著的相关性(P> 0. 05)。结论超声特征中乳腺癌的HER-2表达水平与肿瘤大小和内部钙化具有密切的关系。     

Expression of Cancer-Testis Antigens in Stem Cells: Is it a Potential Drawback or an Advantage in Cancer Immunotherapy

Cancer-testis antigens (CTAs) are a group of tumor associated antigens with a restricted expression patternin normal gametogenic tissues but expression in a broad range of malignancies. Their expression pattern hasmade them potential targets for immunotherapy. However, expression of some of these antigens has beendemonstrated in normal stem cells as well as cancer stem cells (CSCs). As CSCs have been shown to be sourcesof metastasis and tumor recurrence, novel therapies are being focused on their eradication. On the other hand,CTA expression in normal stem cells raises the possibility that CTA based immunotherapies cause side effectsin normal tissues.     

Side Population Cell Level in Human Breast Cancer and Factors Related to Disease-free Survival

Side population (SP) cells have stem cell-like properties with a capacity for self-renewal and are resistantto chemotherapy and radiotherapy. Therefore the presence of SP cells in human breast cancer probably hasprognostic value. Objective: To investigate the characteristics of SP cells and identify the relationship betweenthe SP cells levels and clinico-pathological parameters of the breast tumor and disease-free survival (DFS) inbreast cancer patients. Materials and Methods: A total of 122 eligible breast cancer patients were consecutivelyrecruited from January 1, 2006 to December 31, 2007 at Yunnan Tumor Hospital. All eligible subjects receivedconventional treatment and were followed up for seven years. Predictors of recurrence and/or metastasis andDFS were analyzed using Cox regression analysis. Human breast cancer cells were also obtained from freshhuman breast cancer tissue and cultured by the nucleic acid dye Hoechst33342 with Verapami. Flow cytometry(FCM) was employed to isolate the cells of SP and non-SP types. Results: In this study, SP cells were identifiedusing flow cytometric analysis with Hoechst 33342 dye efflux. Adjusted for age, tumor size, lymph nodal status,histological grade, the Cox model showed a higher risk of recurrence and/or metastasis positively associatedwith the SP cell level (1.75, 1.02-2.98), as well as with axillary lymph node metastasis (2.99, 1.76-5.09), pathologyinvasiveness type (1.7, 1.14-2.55), and tumor volume doubling time (TVDT) (1.54, 1.01-2.36). Conclusions: TheSP cell level is independently associated with tumor progression and clinical outcome after controlling for otherpathological factors. The axillary lymph node status, TVDT and the status of non-invasive or invasive tumorindependently predict the prognosis of breast cancer.     

紫杉醇治疗乳腺癌56例临床观察

目的:观察含紫杉醇的化疗方案治疗乳腺癌的疗效及不良反应。方法:用含紫杉醇的化疗方案治疗乳腺癌56例,紫杉醇剂量(联合化疗135mg/m2,单药化疗175mg/m2)均匀3小时静脉输注,3周~4周重复1次,平均每例给药5次,在紫杉醇化疗前常规给予地塞米松、西米替丁、非拉根预防过敏。结果:总有效率44/56(78.57%)。Ⅱ期25/31(80.65%),Ⅲ、Ⅳ期19/25(76%),主要不良反应为白细胞下降、肌肉关节疼痛、手足麻木、脱发和面色潮红。结论:紫杉醇治疗乳腺癌疗效肯定,不良反应可耐受。     

Aberrant Expression of Markers of Cancer Stem Cells in Gastric Adenocarcinoma and their Relationship to Vasculogenic Mimicry

Background: Gastric cancer is the second leading cause of cancer-related death in Asia, and the majoritytype is gastric adenocarcinoma (GAC). Most GAC patients die of recurrence and metastasis. Cancer stem cells(CSCs) have been thought to be responsible for the initiation, development, metastasis, and ultimately recurrenceof cancer. In this study, we aimed to investigate expression and clinical significance of CSCs markers, CD133 andLgr5, and vasculogenic mimicry (VM) in primary GAC. Materials and Methods: Specimens from 261 Chinesepatients with follow-up were analyzed for CD133, Lgr5 protein expression and VM by immunohistochemicaland histochemical staining. The Pearson Chi’s square test was used to assess the associations among the positivestaining of these markers and clinicopathological characteristics. Postoperative overall survival time was werestudied by univariate and multivariate analyses. Results: In GAC tissues, positive rates of 49.0%, 38.7%, and26.8% were obtained for CD133, Lgr5, and VM, respectively. The mean score of microvessel density (MVD)was 21.7±11.1 in GAC tissues. There was a significantly difference between the positive and negative groups.There was a positive relationship between the VM, the expression of CD133 and Lgr5, and the score of MVDand the grades of tumor, lymph node metastasis, TNM stages (all p<0.05). The overall mean survival time ofthe patients with CD133, Lgr5, VM, and MVD (≥22) positive expression was lower than that of patients withnegative expression. The score of MVD, positive expression of CD133 and VM were independent prognosticfactors of GAC (p<0.05). Conclusions: VM, and expression of CD133, Lgr5, and the score of MVD are relatedto grades of tumor, lymph node metastasis, TNM stages, and overall mean survival time. It is suggested thatCSCs and VM could play an important role in the evolution of GAC.     

乳腺癌中survivin的表达及其与Caspase-7和肿瘤病理特征的关系

 目的 研究乳腺癌组织中survivin的表达及其与Caspase-7和肿瘤病理特征的关系。 方法 免疫组织化学SP法检测survivin和Caspase-7在乳腺癌、 乳腺小叶增生、乳腺导管单纯性增生、乳腺癌旁不典型增生组织中的表达情况,并分析其与 临床病理学特征的关系。 结果 survivin和Caspase-7在乳腺癌中的阳性表达率分别为62.0%和81.7%,两者的表达呈 正相关 (r=10.22,P＜0.01)。在乳腺小叶增生、乳腺导管单纯性增生和乳腺癌旁 不典型增生组织中, survivin阳性表达率分别为0、0、6.67%,明显低于乳腺癌组(P<0.01) ;乳腺 良性病变组中Caspase-7 阳性表达均为100%,明显高于乳腺癌组(P<0.05)。 survivin与肿瘤大小和淋巴结转移有关,与年龄、组织学分级及C-erbB-2表达无关。 结论 survivin蛋白在乳腺癌中表达上调, 且与肿瘤大小和淋巴结转移有关,其可能通过抑制 Caspase-7的活性来抑制凋亡,促使肿瘤增殖。     

乳腺癌组织中PDK4的表达及其与预后的关系

 目的 探讨影响乳腺癌组织中PDK4的表达与预后的相关性。方法 PubMed数据库The Gene Expression Omnibus（GEO, http://www.ncbi.nlm.nih.gov/geo/）分析PDK4在正常乳腺及乳腺癌组织中的表达水平。使用The Kaplan–Meier plotter（www.kmplot.com）分析工具评估PDK4表达水平与乳腺癌临床预后的相关性。TGF-β细胞因子处理MCF-7细胞系,Real-time PCR技术检测经处理后MCF-7细胞系中PDK4 mRNA表达水平。siRNA干扰技术降低细胞内PDK4的表达水平,Transwell检测MCF-7细胞的侵袭能力。结果 PDK4在乳腺癌组织中的表达水平低于正常乳腺组织,并且PDK4低表达与乳腺癌预后差呈正相关;在乳腺癌细胞系中,TGF-β降低PDK4 mRNA表达水平,且PDK4的低表达增强了MCF-7细胞的侵袭能力。结论 PDK4低表达是乳腺癌预后差的一个重要指标。     

IL-24-TRAIL基因载体的构建及其对乳腺癌干细胞迁移和凋亡的影响

目的 构建真核表达载体pIRES-IL-24-TRAIL,探讨其对乳腺癌干细胞迁移和凋亡的影响。方法 以人胎盘组织总RNA为模板,采用RT-PCR二步法,扩增TRAIL的cDNA序列,将其克隆入pIRES载体,扩增IL-24的cDNA序列,将其克隆入pIRES- TRAIL载体,构建重组真核表达载体pIRESIL-24- TRAIL,以Lipofectamine2000转染技术,将质粒pIRES- IL-24-TRAIL导入乳腺癌干细胞MCF-7和MDA-MB-231,流式细胞仪检测各组细胞凋亡情况,划痕实验比较不同组间细胞迁移情况。结果（1）克隆到TRAIL、IL-24基因的cDNA序列,成功构建其真核表达载体。（2）流式细胞仪发现,实验组比对照组细胞凋亡率高(P<0.05)。（3）细胞划痕实验显示实验组细胞迁移能力明显低于对照组（P＜0.05）。结论 pIRES-IL-24-TRAIL构建成功,并能诱导乳腺癌干细胞凋亡、降低其迁移能力。     

The Relationship Between Expression of EpCAM Cancer Stem Cell Marker with Histopathological Grading,Lymphovascular Invasion,and Metastases in Colorectal Adenocarcinoma

Objective: The aim of this study was to analyze the expression of EpCAM in the colorectal adenocarcinoma. Material and methods: This study used a cross-sectional design. One hundred and thirteen paraffin embedded block of Colorectal Adenocarcinoma were assessed using anti-EpCAM/Epithelial Specific Antigen (Ber-EP4) mouse monoclonal antibody and their expression were performed using Olympus CX-43 light microscope. The relationship between EpCAM expression with histopathological grade of colorectal adenocarcinoma, lymphovascular invasion and metastases ability were statistically analyzed by Chi-Square tests and presented in tables using SPSS 18. Results: From 113 samples, in samples with lymphovascular invasion there were 37 samples (32.7%) with strong expression, while those with weak expression were 19 samples (16.8%). There were 39 samples with metastases and strong expression of EpCAM (34.5%), while 21 samples with weak expression (18.6%). There was a significant relationship between the expression of cancer stem cell marker EpCAM with lymphovascular invasion and colorectal adenocarcinoma metastases (p = 0.002), but there was no significant relationship with histopathological grade (p = 0.574). Conclusion: The EpCAM expression can be used as a prognostic factor, and can be considered as a predictive or an option for target therapy in colorectal adenocarcinoma.