肿瘤患者外周血淋巴细胞化疗药物敏感性检测的应用研究

目的：探讨肿瘤患者外周血淋巴细胞对临床常用化疗药物的敏感性,筛选个体敏感的化疗药物,指导临床用药。方法：采用MTT法检测154例肿瘤患者外周血淋巴细胞对常用15种化疗药物的敏感性。结果：本方法测定的结果与临床常规化疗的结果基本一致。结论：MTT法是一种简便、快速、可行的体外药敏实验方法,可为肿瘤患者化疗提供重要的实验依据。     

DNA依赖蛋白激酶与大肠癌淋巴转移相关性研究

Objective:To investigate DNA-dependent protein kinase(DNA-PK)expression,and its relationship with lymphatic metastasis in colorectal cancer.Methods:Tumor tissues from 60 patients,divided into two groups according to lymphatic metastasis,were immunohistochemically stained to detect the DNA-PK expression including Ku70,Ku80 and PKcs proteins.Results:Positivity of both Ku70 and Ku80 in colorectal cancer was negatively correlated with lymphatic metastasis with an rvalue of-0.57 and -0.38,respectively.Similar correlation was found between Ku expression,especially Ku70.and long-term survival.PKcs,however,displayed no significant correlation.Statistical analysis failed to detect any correlation between DNA-PK expression,and clinical characteristics,such as age,sex,tumor Jocation,tumor thickness and distant metastasis(P＞0.05).Conclusion:DNA-PK expression,especially Ku70 expression,is negatively correlated with lymphatic metastasis,and the survival of patients with colorectal cancer.Ku70 expression may be a potential indicator for the preoperative evaluation,and prognosis in colorectal cancer.     

Investigation of the relationship between DNA-dependent protein kinase and lymphatic metastasis in colorectal cancer

Objective  To investigate DNA-dependent protein kinase (DNA-PK) expression, and its relationship with lymphatic metastasis in colorectal cancer. Methods  Tumor tissues from 60 patients, divided into two groups according to lymphatic metastasis, were immunohistochemically stained to detect the DNA-PK expression including Ku70, Ku80 and PKcs proteins. Results  Positivity of both Ku70 and Ku80 in colorectal cancer was negatively correlated with lymphatic metastasis with an r value of −0.57 and −0.38, respectively. Similar correlation was found between Ku expression, especially Ku70, and long-term survival. PKcs, however, displayed no significant correlation. Statistical analysis failed to detect any correlation between DNAPK expression, and clinical characteristics, such as age, sex, tumor location, tumor thickness and distant metastasis (P > 0.05). Conclusion  DNA-PK expression, especially Ku70 expression, is negatively correlated with lymphatic metastasis, and the survival of patients with colorectal cancer. Ku70 expression may be a potential indicator for the preoperative evaluation, and prognosis in colorectal cancer. Supported by a grant from the Scientific Research Project of the Bureau of Health of Jiading in Shanghai (No. KYXM-2004-11-07).     

The association of DNA-dependent protein kinase activity with chromosomal instability and risk of cancer

The DNA double-strand breaks (DSBs) repair pathway has been implicated in maintaining genomic integrity via suppression of chromosomal rearrangements. DNA-dependent protein kinase (DNA-PK) has an important role with DNA DSBs repair. In this study, 93 of untreated cancer patients and 41 of cancer-free healthy volunteers were enrolled. Peripheral blood was collected, separated and centrifuged; DNA-PK activity was measured by DNA-pull-down assay. The expressions of DNA-PKcs, Ku70 and Ku86 were examined by RT-PCR assay and western blotting. Chromosomal aberrations were examined by cytogenetic methods. DNA-PK activities of peripheral blood lymphocytes (PBL) in patients with uterine cervix or breast cancer were significantly lower than those in normal volunteers. Age and smoking had no association with DNA-PK activity, whereas DNA-PK activity and the expression of Ku70, Ku86 and DNA-PKcs in RT-PCR were interrelated. A similar tendency was seen in western blot assay but less clear than in RT-PCR. Therefore, the association between DNA-PK activity and expression of DNA-PK in protein level could not be concluded. The frequency of chromosome aberration, such as dicentric chromosomes and excess fragment increased as the DNA-PK activity decreased. In conclusion, DNA-PK activity is associated with chromosomal instability. DNA-PK activity in PBL is associated with risk of breast and uterine cervix cancer. DNA-PK activity in PBL can be used to select individuals for whom an examination should be performed because of their increased susceptibility to breast and uterine cervix cancer.     

卵巢癌患者外周血T淋巴细胞的表达分析

目的探讨卵巢癌患者外周血T淋巴细胞的表达变化情况,分析CD+4CD+25表达与预后的关系。方法选择2009年2月到2012年11月收治的35例卵巢癌患者作为试验组,另选择35例同期体检排除肿瘤及免疫相关性疾病的健康人作为对照组,检测两组患者外周血中T淋巴细胞的表达水平。试验组同期分离肿瘤淋巴细胞进行免疫染色分析。结果对照组和试验组外周血淋巴细胞中CD+4CD+25的表达量分别为(5.36±0.63)%和(44.12±6.23)%,差异有统计学意义(P<0.05)。外周血淋巴细胞CD+4CD+25比例与卵巢癌患者年龄、NCCN分期、组织学类型和淋巴结转移均无关(P>0.05)。CD+4CD+25高表达组卵巢癌患者总生存期低于CD+4CD+25低表达组(P<0.05)。结论卵巢癌患者外周血CD+4CD+25淋巴细胞多呈现高表达,与预后密切相关,可成为卵巢癌预后指标和免疫治疗的生物靶点。     

Altered mode of allelic replication accompanied by aneuploidy in peripheral blood lymphocytes of prostate cancer patients

Replication timing of the genetic material is a highly programmed process correlated with expression, stability and methylation capacity. An important aspect of that timing is the temporal order of allelic replication: a synchronous mode for biallelically expressed genes and an asynchronous for monoallelically expressed genes. Previous studies showed that malignancy is associated with changes in the inherent mode of allelic replication, and even normal cells of cancer patients display alterations in the replication of various genes. Using fluorescence in situ hybridization (FISH), we checked whether allelic-replication mode differentiates cancer patients from healthy individuals. We focused on prostate cancer (CAP), the most common diagnosed cancer and the second leading cause of cancer death in men over 50 years old. Five nonrelated genes and a nontranscribed DNA sequence associated with chromosomal segregation were used in our study. All 6 tested loci displayed in peripheral blood lymphocytes stimulated with phytohemagglutinin (PHA) of CAP patients loss of their inherent temporal order of allelic replication, coupled with aneuploidy, the outcome of chromosome malsegregation. The replication-timing modification is a reversible epigenetic alteration, evidenced by our ability to resurrect the normal pattern in all 6 tested loci by introducing an inhibitor of methyl transferase. On the other hand, the methylation-blocking agent failed to obliterate aneuploidy. The replication alteration accompanied by aneuploidy, detected in peripheral blood cells, distinguishes between CAP patients and individuals with benign prostate hyperplasia (BPH; a common disorder in elderly men) better than the routinely used blood marker, the prostate-specific antigen (PSA).     

肺癌患者外周血淋巴细胞与癌细胞体外化疗药敏实验研究

目的　研究肺癌患者外周血淋巴细胞和癌细胞体外化疗药敏的相关性。方法　采用MTT法测定了 74例肺癌患者外周血淋巴细胞及其癌细胞对 15种临床常用化疗药物的敏感性。结果　肺癌患者外周血淋巴细胞与癌细胞对其中 12种化疗药的敏感性有很好的相关性 ,差异无显著性 (P >0 .0 5 )。结论　外周血淋巴细胞的体外化疗敏感性检测对肺癌的临床化疗药物的选择具有重要的参考价值。     

Lymphokine-activated killer cell activity of peripheral blood,spleen, regional lymph node,and tumor infiltrating lymphocytes in gastric cancer patients

Lymphokine-activated killer (LAK) cell activity of peripheral blood mono-nuclear cells (PBM), spleen cells (SPC), regional lymph node cells (LNC), and tumor-infiltrating lymphocytes (TIL), induced by activation with in-terleukin 2 (IL 2) for 4 days, was evaluated in patients with gastric carcinoma. TIL exhibited the lowest LAK activity and the cytotoxicity of LNC was significantly lower than that of either PBM or SPC. There was no difference between PBM and SPC. Then, there were significant correlations of LAK activity among PBM, SPC, and LNC, whereas poor correlations were observed in the cytotoxicity between TIL and PBM, SPC, or LNC. Phenotypic analysis of each cell population was performed before and after activation with IL 2. Before culture, the cells mediating natural killer (NK) activity such as CD16⁺, CD56⁺, and CD57⁺ cells were few in LNC and TIL. However, CD56⁺ and CD57⁺ cells in TIL were increased after culture. Then, CD4⁺ Leu8⁺ and CD8⁺ CD11⁺ cells, which identify suppressor cell function, were not elevated in LNC or TIL, as compared to that in PBM or SPC. Further, the proportions of OKIa1⁺ and CD25⁺ cells expressing T-cell activation and IL 2 receptor were uniformly increased in all cell populations after culture. These results indicate the differential reactivity of each lymphocyte population to IL 2 and fundamental dysfunction of LNC and, especially TIL, suggesting the specific influence of the local tumor environment on the lymphocyte function in the area in patients with gastric carcinoma. © Wiley-Liss, Inc.     

ECT2 mRNA及蛋白在胃癌患者外周血中的表达及意义

目的:探讨ECT2 mRNA及蛋白在胃癌患者外周血中的表达水平及其意义。方法:应用逆转录聚合酶链式反应（RT-PCR）及酶联免疫吸附实验（ELISA）测定50例胃癌患者及30例健康志愿者外周血ECT2 mRNA及蛋白的表达水平。结果:胃癌患者外周血中ECT2 mRNA阳性表达率为54%,健康志愿者外周血中ECT2 mRNA均阴性表达(P<0.05)。ECT2蛋白在胃癌患者外周血中的浓度（1 306.389±215.824)ng/L显著高于健康志愿者外周血中的浓度（502.718±69.440)ng/L(P<0.05),并且与TNM分期、淋巴结转移和浸润深度相关(P<0.05)。结论:外周血中ECT2 mRNA及蛋白的表达与胃癌关系密切,可作为反映胃癌发生、发展过程的有效分子指标。     

外周血红细胞分布宽度、超氧化物歧化酶与胃癌预后的相关性

目的:验证红细胞分布宽度(red blood cell distribution width,RDW)和超氧化物歧化酶(superoxide dismutase,SOD)与初治胃癌患者的临床病理特征及预后的关系.方法:回顾性分析了2015年01月至2017年07月之间200例初治胃癌患者和110例胃息肉患者的临床数据,...     

Prognostic value of the density of tumor-infiltrating lymphocytes in colorectal cancer liver metastases

Tumor-infiltrating lymphocytes (TILs) have been reported to reflect the anti-tumor immune status of patients and to be correlated with their prognosis and therapeutic outcomes. However, the characteristics of the local immune status in metastatic tumors is poorly understood, as primary tumors have been the focus in most previous studies. In addition, the local immune status may be influenced by preoperative chemotherapy. The present study aimed therefore to investigate the relationship between the degree of TIL infiltration and the prognosis in patients with curative resection of colorectal cancer liver metastases and to examine the effects of preoperative chemotherapy on the function of immune cells. A total of 108 patients who underwent curative resection of colorectal cancer liver metastases in our department between May 1996 and January 2017 were enrolled in the present study. Peripheral blood samples were obtained within two weeks before surgery. TIL infiltration was evaluated by immunohistochemical staining of surgically resected specimens of liver metastases using anti-CD8/CD3 antibodies. The mean number of TILs in five different fields was calculated, and patients were classified into a high-TIL group and a low-TIL group. Furthermore, patients were divided into three groups as follows: i) A group of patients who did not receive preoperative chemotherapy; ii) a group of patients who received short-term preoperative chemotherapy for <6 months; and iii) a group of patients who received long-term preoperative chemotherapy for ≥6 months. The results demonstrated that the density of TILs in colorectal liver metastases was not correlated with the absolute peripheral lymphocyte count in all patients. Furthermore, the degree of CD8⁺TIL infiltration in liver metastases was significantly lower in the recurrence group compared with the recurrence-free group following hepatectomy. In all patients with colorectal liver metastases, the degree of CD8⁺TIL infiltration was significantly associated with the relapse-free and overall survival. In patients without preoperative chemotherapy, the degree of CD8⁺TIL infiltration was significantly associated with the relapse-free survival, and a high CD8⁺TIL presence tended to have a better effect on the overall survival than a low CD8⁺TIL presence. In the short-term chemotherapy group, the degree of CD8⁺TIL infiltration was significantly associated with the relapse-free and overall survival. In the long-term chemotherapy group, there were no significant differences between the high- and low- CD8⁺TIL groups in the relapse-free and overall survival. In contrast to CD8⁺TILs, CD3⁺TILs showed a poor prognostic ability. In summary, the degree of CD8⁺TIL infiltration in colorectal cancer liver metastases may be correlated with patient prognosis. However, in patients who received long-term chemotherapy before surgery, the degree of TIL infiltration was not necessarily associated with prognosis as the anti-tumor effects of TILs may decrease. The degree of CD8⁺TIL infiltration may therefore be considered as a useful prognostic factor in patients with colorectal liver metastases, but the prognostic accuracy may decrease in patients who received long-term chemotherapy.     

Phytohaemagglutinin responsiveness of peripheral lymphocytes and survival in patients with primary breast cancer

Summary The response of peripheral blood lymphocytes from 153 breast cancer patients to the T-cell mitogen phytohaemagglutinin (PHA) was measured prior to surgery and patients assigned to either high (Hi/PHA) or low (Lo/PHA) PHA response groups. Patients were followed up for between 8 and 12 years, or until death. Clinical characteristics were similar in Hi/PHA and Lo/PHA groups. In the 96 patients who received no postoperative adjuvant therapy, overall and relapse-free survival was significantly longer in Hi/PHA than in Lo/PHA patients (p < 0.005, and p < 0.025 respectively); relative risk of death within 12 years was 0.52 and of relapse was 0.64. In 47 patients who were treated with radiotherapy postoperatively, there was no correlation between the PHA response and overall or relapse-free survival. The implications of these findings for the selection of breast cancer patients for adjuvant treatment following surgery are discussed.     

外周血单个核细胞GTSE1水平与上皮性卵巢癌预后的相关性

目的:探究外周血单个核细胞G₂/S期应答相关蛋白1(G₂ and S phase-expressed-1,GTSE1)水平与上皮性卵巢癌(epithelial ovarian cancer, EOC)预后的关系。方法:GEPIA和Kaplan-Meier Plotter分析GTSE1在卵巢癌组织中的表达及其与预后的关系。选择2017年04月至2020年10月本院126例EOC患者(EOC组)和40例同期无EOC的健康体检者(对照组)作为研究对象。Ficoll密度梯度离心法分离外周血中单个核细胞,蛋白质免疫印迹法检测GTSE1水平。Spearman相关分析EOC组织和外周血单个核细胞中GTSE1水平的关系。随访了解EOC预后,用COX回归分析EOC预后的风险因素。结果:GEPIA和Kaplan-Meier Plotter分析结果显示,卵巢癌组织中GTSE1水平高于正常卵巢组织(P<0.05),GTSE1低水平卵巢癌患者的中位无进展生存时间高于GTSE1高水平卵巢癌患者(P<0.05)。随访期间复发98例(77.8%),其中铂类敏感型...     

术前外周血中性粒细胞与淋巴细胞比值评估肝细胞癌根治性切除术后预后的临床价值

目的 探讨肝细胞癌(HCC)患者术前外周血中性粒细胞与淋巴细胞的比值(NLR)对根治性切除术后预后评估的临床价值.方法 收集行HCC根治性切除术的161例HCC患者的临床资料,根据患者术前NLR值情况,将患者分为高NLR组(NLR≥2.60,n=52)及低NLR组(NLR﹤2.60,n=109).应用Kaplan-Meier方法分析患者的生存率,并采用Log-rank检验差异;采用Cox回归模型分析影响HCC患者总体生存率的预后因素.结果161例HCC患者的中位随访时间为29个月,高NLR组的1、3、5年总体生存率分别为79.6%、70.8%、41.1%,中位肿瘤复发时间14.9个月;低NLR组的1、3、5年总体生存率分别为90.4%、79.6%、48.7%,中位肿瘤复发时间为20.1个月,高NLR组的1、3、5年总体生存率均低于低NLR组(P﹤0.05),中位肿瘤复发时间短于低NLR组(P﹤0.05);单因素分析结果显示:肿瘤包膜、TNM分期、术前AFP、术前高NLR、淋巴结转移、肿瘤大小与HCC根治性切除术后预后生存时间有关,差异有统计学意义(P﹤0.05);Cox多因素分析结果显示:TNM分期为Ⅲ~Ⅳ期、术前AFP﹤400 ng/ml、术前NLR≥2.60、淋巴结转移、肿瘤大小≥10 mm为影响HCC患者根治性切除术后预后的独立危险因素(P﹤0.001).结论 术前NLR是影响HCC患者根治性切除术后预后的独立危险因素,可作为评估患者预后的指标,术前高NLR者其预后较差.     

肺癌患者外周血免疫细胞毒细胞的检测

 目的 探讨肺癌患者群体的免疫细胞毒细胞的功能状态。方法 应用流式细胞仪测定30例手术前肺癌患者和30例该院健康职工外周血总T淋巴细胞、自然杀伤细胞（NK）、NK样T杀伤细胞（NKT）、γδT细胞和CTL细胞。结果 γδT细胞：正常组为6.54±2.94，患者组为3.58±2.14，t = 3.730，P =0.001。CTL细胞：正常组为17.21±4.49，患者组为10.86±4.67，t =5.130，P＜0.001；其他被测细胞的百分率，经统计学分析，均无统计学意义。结论 特异性免疫杀伤细胞低下对肺癌的发生有重要影响。非特异性免疫细胞毒细胞影响有限。     

EXPRESSION OF IL-2R ON PERIPHERAL BLOOD LYMPHOCYTES OF PATIENTS WITH COLORECTAL CANCER AND ITS CLINICAL SIGNIFICANCE

In recent years it has been noted that the levels of serum soluble interleukin-2 receptor (sIL-2R) in patients with malignancy were abnormally elevated and the levels of sIL-2R might be considered as an index to monitor patient's condition and assess treatment effects. But in medical literature it was reported that the levels of membrane interleukin-2 receptor (CD25) in peripheral blood lymphocytes (PBL) in patients with malignancy were various, some were elevated, some were dropped.[1] Th…     

Telomere content correlates with stage and prognosis in breast cancer

SummaryPurpose To evaluate the hypothesis that telomere DNA content (TC) in breast tumor tissue correlates with TNM staging and prognosis.Experimental design Slot blot assay was used to quantitate TC in 70 disease-free normal tissues from multiple organ sites, and two independent sets of breast tumors containing a total of 140 samples. Non-parametric Rank–Sums tests, logistic regression and Cox proportional hazards models were used to evaluate the relationships between TC and tumor size, nodal involvement, TNM stage, 5-year survival and disease-free interval.Results TC in 95% of normal tissues was 75–143% of that in the placental DNA standard, whereas only 50% of tumors had TC values in this range. TC was associated with tumor size (p=0.02), nodal involvement (p<0.0001), TNM stage (p=0.004), 5-year overall survival (p=0.0001) and 5-year disease-free survival (p=0.0004). A multivariable Cox model was developed using age at diagnosis, TNM stage and TC as independent predictors of breast cancer-free survival. Relative to the high TC group (>123% of standard), low TC (<101% of standard) conferred an adjusted relative hazard of 4.43 (95% CI 1.4–13.6, p=0.009). Receiver operating characteristic curves using thresholds defined by the TC distribution in normal tissues predicted 5-year breast cancer-free survival with 50% sensitivity and 95% specificity, and predicted death due to breast cancer with 75% sensitivity and 70% specificity.Conclusions TC in breast cancer tissue is an independent predictor of clinical outcome and survival interval, and may discriminate by stage.Colleen A. Fordyce and Christopher M. Heaphy contributed equally to this study     

The prognostic influence of tumour-infiltrating lymphocytes in cancer: a systematic review with meta-analysis

Background:

Tumour-infiltrating lymphocytes (TILs) are often found in tumours, presumably reflecting an immune response against the tumour. We carried out a systematic review and meta-analysis, aiming to establish pooled estimates for survival outcomes based on the presence of TILs in cancer.

Methods:

A Pubmed and Embase literature search was designed. Studies were included, in which the prognostic significance of intratumoural CD3+, CD4+, CD8+, and FoxP3+ lymphocytes, as well as ratios between these subsets, were determined in solid tumours.

Results:

In pooled analysis, CD3+ TILs had a positive effect on survival with a hazard ratio (HR) of 0.58 (95% confidence interval (CI) 0.43–0.78) for death, as did CD8+ TILs with a HR of 0.71 (95% CI 0.62–0.82). FoxP3+ regulatory TILs were not linked to overall survival, with a HR of 1.19 (95% CI 0.84–1.67). The CD8/FoxP3 ratio produced a more impressive HR (risk of death: HR 0.48, 95% CI 0.34–0.68), but was used in relatively few studies. Sample size and follow-up time seemed to influence study outcomes.

Conclusion:

Any future studies should be carefully designed, to prevent overestimating the effect of TILs on prognosis. In this context, ratios between TIL subsets may be more informative.     

86例胃癌前哨淋巴结转移的临床分析及其预后

目的分析胃癌患者前哨淋巴结转移检出的临床意义及其对预后的影响。方法胃癌术中使用亚甲蓝染色定位前哨淋巴结,观察染色后的显影范围和特点,同时对病灶进行活检;分析前哨淋巴结转移检出情况与疾病T分期和发病部位的关系。结果 86例胃癌患者中,T1期的前哨淋巴结转移检出的灵敏性、准确性及假阴性率优于T2期和T3期,差异具有统计学意义（P〈0.0167）。胃下部病灶组的前哨淋巴结转移检出的灵敏性、准确性及假阴性率均优于胃上部和胃中部病灶组,差异具有统计学意义（P〈0.0167）。术后五年内总死亡率为24.42%（21/86例）。结论胃癌术中注射亚甲蓝定位前哨淋巴结有助于确定肿瘤周围淋巴结切除的合理范围,尤其适用于早期胃癌患者,值得临床推广。     

NSCLC外周血循环肿瘤细胞水平检测及其与TNM分期、预后的关系

目的:分析非小细胞肺癌（non-small cell lung cancer,NSCLC）外周血循环肿瘤细胞（circulating tumor cells,CTC）表达特点及其与国际抗癌联盟肿瘤通用分期（tumor node metastasis,TNM）及预后的关系。方法:采集我院胸外科收治的NSCLC患者82例,选取肺良性疾病30例作为良性组及30名正常志愿者作为对照组,入院后均完成外周血CTC检测,比较NSCLC外周血CTC与良性组、对照组之间的差异;比较不同病理类型、TNM分期、不同预后NSCLC患者CTC阳性率,筛选NSCLC预后影响因素。结果:NSCLC组CTC阳性率高于良性组与对照组（P＜0.05）,良性组、对照组组间对比差异无统计学意义（P＞0.05）;不同病理类型NSCLC患者CTC阳性率比较差异无统计学意义（P＞0.05）;不同TNM分期NSCLC患者CTC阳性率比较差异有统计学意义（P＜0.05）,各分期CTC阳性率比较:Ⅲa期＞Ⅱ期＞Ⅰ期;CTC阴性NSCLC患者随访中位无 病生存时间（disease free survival,DFS）长于阳性组,复发、转移率低于阳性组（P＜0.05）;有吸烟史、CTC阳性、高TNM分期、伴淋巴结转移的NSCLC患者中位DFS低于无吸烟史、CTC阴性、低TNM分期、无淋巴结转移的NSCLC患者（P＜0.05）;Cox回归分析显示:TNM分期、CTC、淋巴结转移均为影响NSCLC患者中位DFS的相关因素（P＜0.05）。结论:NSCLC外周血CTC阳性率较高,且与TNM分期、淋巴结转移共同影响患者预后。