TS基因3’-UTR多态性与晚期肺腺癌患者对培美曲塞敏感度的关系

目的 探讨胸苷酸合成酶( thymidylate synthase,TS)基因3’端非翻译区(untranslated region,UTR)1 494 bp处6 bp缺失或插入多态(-6 bp或+6 bp)与晚期肺腺癌患者接受培美曲塞方案化疗敏感度的关系.方法 采用AollgTM探针结合实时荧光PCR技术检测接受培美曲塞方案化疗的106例晚期肺腺癌患者外周血中TS基因型,分析携带TS不同基因型患者与化疗敏感度的关系.结果 (1)106例晚期肺腺癌患者中TS基因3’-UTR 1 494 bp处6 bp缺失或插入多态基因频率分别为:-6 bp/-6 bp55.7％(59/106),-6 bp/+6 bp 44.3％(47/106),+6 bp/+6 bp 0(0/106);(2)总体近期有效率(CR+PR)为23.58％,携带TS基因3’-UTR 1494 bp处-6 bp/-6 bp基因型化疗有效率是携带-6 bp/+6 bp基因型有效率的4.382倍(95％ CI:1.462～13.130,P=0.008);(3)携带-6 bp/-6 bp基因型与携带-6 bp/+6 bp基因型患者的中位PFS、中位OS差异均具有统计学意义(3.400月vs.2.477月,p=0.001;14.239月vs.12.194月,P=0.000);提示携带-6 bp/-6 bp基因型患者在PFS、OS具有显著优势.(4)Cox多因素分析临床病理因素与生存时间的关系得出年龄越大,病理分期越晚,携带TS3’-UTR 1 494位点处-6 bp/+6 bp基因型患者的死亡风险越大.结论 TS基因3’- UTR 1 494 bp处6 bp缺失或插入多态(-6 bp或+6 bp)可能与晚期肺腺癌患者培美曲塞方案化疗敏感度相关,可作为晚期肺腺癌患者个体化治疗预测因子之一.     

TS基因3′-UTR多态性与晚期肺腺癌患者对培美曲塞敏感度的关系

 目的
探讨胸苷酸合成酶(thymidylate synthase,TS）基因3′端非翻译区(untranslated region,UTR)1 494 bp处6 bp缺失或插入多态(-6 bp或+6 bp)与晚期肺腺癌患者接受培美曲塞方案化疗敏感度的关系。方法采用AollgTM探针结合实时荧光PCR技术检测接受培美曲塞方案化疗的106例晚期肺腺癌患者外周血中TS基因型,分析携带TS不同基因型患者与化疗敏感度的关系。结果（1）106例晚期肺腺癌患者中TS基因3′- UTR 1 494 bp处6 bp缺失或插入多态基因频率分别为:-6 bp/-6 bp 557%（59/106）,-6 bp/+6 bp 44.3%（47/106）,+6 bp/+6 bp 0（0/106）;（2）总体近期有效率(CR + PR)为23.58%,携带TS基因3′-UTR 1494 bp处-6 bp/- 6 bp基因型化疗有效率是携带-6 bp/+6 bp基因型有效率的4.382倍 (95% CI:1.462～13.130,P=0.008);（3）携带-6 bp/-6 bp基因型与携带-6 bp/+6 bp基因型患者的中位PFS、中位OS差异均具有统计学意义（3.400月vs.2.477月,P=0001;14.239月vs.12.194月,P=0.000）;提示携带-6 bp/-6 bp基因型患者在PFS、OS具有显著优势。（4）Cox多因素分析临床病理因素与生存时间的关系得出年龄越大,病理分期越晚,携带TS3′- UTR 1 494位点处-6 bp/+6 bp基因型患者的死亡风险越大。结论TS基因3′- UTR 1 494 bp处6 bp缺失或插入多态(-6 bp或+6 bp)可能与晚期肺腺癌患者培美曲塞方案化疗敏感度相关,可作为晚期肺腺癌患者个体化治疗预测因子之一。     

晚期非小细胞肺癌循环肿瘤细胞的测定及其意义

摘 要：［目的］ 分析晚期非小细胞肺癌患者外周血循环肿瘤细胞（circulating tumor cells,CTCs）与临床特征、肿瘤标志物间的相关性。［方法］ 收集2016年7月至2017年4月徐州医科大学第二附属医院经病理证实的36例晚期非小细胞肺癌患者的临床、病理等资料以及10位健康志愿者。所有研究对象均行外周血循环肿瘤细胞检测,采用免疫磁珠阴性富集技术分离CTCs,利用荧光原位杂交和免疫荧光染色鉴别CTCs,在荧光显微镜下观察并统计CTCs。［结果］ 36例非小细胞肺癌患者的循环肿瘤细胞水平明显高于健康志愿者（P=0.000）。单因素分析结果显示,晚期非小细胞肺癌外周血CTCs与临床分期(P=0.010)、远处转移(P=0.006)以及远处转移个数(P=0.018 )相关。多因素回归分析结果则显示,仅远处转移与 CTCs 的阳性表达具有相关性(P=0.011)。CTCs阳性表达与CEA、SCCA、NSE无关( P＞0．05),但CYFRA21-1组的CTCs阳性率明显高于正常组(P=0.034)。［结论］ CTCs在晚期非小细胞肺癌中的表达与远处转移密切相关,且CTCs与传统肿瘤标志物的联合应用可为疾病复发和转移提供依据。     

围手术期血循环中肺癌细胞与预后的关系

[目的]分析可手术性非小细胞肺癌 (NSCLC)患者术前和术中癌细胞血道播散及其对患者预后的影响。[方法]术前或术中采集外周血或肺静脉血 ,应用流式细胞仪进行癌细胞定量测定。患者于手术后定期随访 ,并与检测结果作统计学分析。[结果]对44例NSCLC患者血样进行了检测 ,共发现16例存在癌细胞 ,阳性率为36.4 % ,癌细胞平均含量为0.286×106/L。其中60岁以下年龄组的癌细胞阳性率显著高于60岁以上年龄组(P<0.05) ,鳞癌患者的癌细胞含量显著高于腺癌患者 (P<0.05)。但25%的腺癌患者在术前已出现癌细胞播散 ,术中阳性率为42.9%,癌细胞阳性率与病期无明显相关性 ;鳞癌患者术前未发现阳性病例 ,术中阳性率为57.1% ,阳性者以Ⅲ、Ⅳ期为主。随访结果显示 ,血循环癌细胞阳性和阴性组的中位生存期分别为15个月和>30个月 ,2年生存率分别为31.3%和71.4 % ,具有统计学显著差异 (P<0.05)。[结论]肺癌患者在手术前或手术过程中可发现血液中存在癌细胞 ,此类患者的预后差、生存期短。     

肿瘤患者循环血管内皮干/祖细胞的检测及体外诱导分化

目的　证实外周血中循环血管内皮细胞 (CECs)及血管内皮前体细胞 (CEPs)的存在 ,检测内皮前体细胞的增殖分化能力。方法　应用流式细胞术检测 5 7例肿瘤患者及 15例正常人外周血中的CECs和CEPs ,应用ELISA方法检测血清VEGF水平。选择经造血干细胞动员的肿瘤患者 ,分离单个核细胞 (MNCs) ,在VEGF、IGF及bFGF条件下培养血管内皮细胞 ,并应用透射电镜法对培养细胞进行鉴定。结果　肿瘤患者外周血中CECs、CEPs含量分别为 0 .3 78%± 0 .0 47%、0 .0 5 9%± 0 .0 13 % ,高于正常对照组 ,且与血清VEGF水平相关。透射电镜下培养细胞可见Weibel-Palade小体 ,为血管内皮细胞的特异性标志。结论　外周血中存在CECs和CEPs ,在肿瘤患者体内存在血管内皮及其前体细胞的动员 ,循环血管内皮干 /祖细胞可进一步分化成熟     

Expression of Thymidylate Synthase in Human Non-small Cell Lung Cancer

5-Fluorouracil (5-FU) has been used worldwide, and the correlation between its effects and thymidylate synthase (TS) expression has been reported in gastrointestinal malignancy. But the significance of TS expression for 5-FU-based chemotherapy has rarely been reported in non-small cell lung cancer (NSCLC). We investigated surgically resected specimens of 23 consecutive patients with previously untreated NSCLC. We used immunohistochemistry and western blot analysis with anti-TS polyclonal antibody to evaluate the existence of TS, and fluorodeoxyuridine-5'-monophosphate (FdUMP) binding assay to evaluate the enzymic activity of TS. We found that 14 samples (60.9%) were positive immunohistochemically, and that the results of immunohistochemistry closely reflected the enzymic activity measured by FdUMP binding assay (ranging from 1.8 to 56.9 pmol/g protein). These results seem to support our experience that 5-FU and its derivatives are clinically significantly effective as a postoperative adjuvant chemotherapy against NSCLC.     

肺癌患者外周血循环肿瘤细胞检测的研究进展

肺癌是我国发病率和死亡率均较高的恶性肿瘤,预后较差.患者5年生存率还有待进一步提高,患者致死的主要原因是肺癌的复发和转移.血行播散是肺癌转移的重要途径,肿瘤细胞可以通过血液循环发生远处转移.肿瘤细胞进入血液循环是肿瘤发生远处转移的关键步骤之一,因此对肺癌患者循环肿瘤细胞检测的研究已受到越来越多的重视.在此,我们就相关研究进展加以概述.     

巢式RT-PCR检胃癌患者外周血循环癌细胞的研究

目的　分析胃癌患者外周血中CK 2 0mRNA及CEAmRNA的异常表达及意义。方法　采用RT PCR方法检测 5 2例胃癌患者术前外周血中CK 2 0mRNA及CEAmRNA的表达。结果　胃癌患者外周血中CK 2 0mRNA、CEAmRNA阳性表达率分别为 42 .3 % (2 2 /5 2 )及 5 1.9% (2 7/5 2 ) ,CK 2 0mRNA和CEAmRNA两者中至少 1个基因为阳性的表达率为 63 .5 % (3 3 /5 2 )。胃癌患者外周血中CK 2 0mRNA表达与胃癌的远处转移有关 ,CEAmRNA表达、CK 2 0mRNA和CEAmRNA都表达、CK 2 0mR NA和CEAmRNA两者中至少 1个表达与胃癌的浸润深度、淋巴结转移和远处转移相关 (P <0 .0 5 )。结论　胃癌患者外周血CK 2 0和CEA的RT PCR检测有助于患者预后的判断 ;联合检测CK 2 0mRNA和CEAmRNA可增加外周血循环癌细胞检测的敏感性     

乳腺癌患者外周血循环RNA检测研究进展

乳腺癌患者外周血循环RNA具有和乳腺癌相一致的分子变化,与传统的乳腺癌检测方法相比,循环RNA的检测在诸多方面具有一定优势。通过分子生物学技术检测循环RNA,有助于提供乳腺癌的早期诊断、疗效观察、临床治疗方案的选择及预后监测的相关信息,具有良好的临床应用前景。     

肺癌患者外周血中CK19-mRNA表达的检测

目的　检测肺癌患者外周血中CK 19 mRNA的表达情况。方法　采用逆转录PCR (RT PCR )方法 ,分别检测有明确肿瘤病灶的肺癌患者、肺部良性疾病患者和健康对照者外周血中CK 19 mRNA的表达。结果　 70例肺癌患者外周血中CK 19 mRNA表达阳性率为 3 7.14 % ,3 8例肺部良性疾病患者阳性率为 18.42 % ,3 7例健康对照者阳性率为 5 .41% ,肺癌组与肺部良性疾病组、健康对照组间阳性率均有显著性差异 (P <0 .0 5 ) ,CK 19 mRNA表达阳性率与肿瘤分期及病理类型无相关性。结论　外周血中CK 19 mRNA作为肺癌的 1项分子检测指标 ,具有一定的临床应用价值 ,值得进一步扩大样本研究及随访 ,其检测的灵敏度及特异度尚有待提高     

循环肿瘤细胞检测在预测非小细胞肺癌化疗疗效中的应用

目的 探讨循环肿瘤细胞检测在预测非小细胞肺癌化疗疗效中的应用.方法 选择非小细胞肺癌患者(肺Ca组)共25例,另选择慢性阻塞性疾病患者(COPD组)25例作为对比研究.入院后及化疗2个周期后分别采集所有参与研究的人员静脉血7.5 ml,利用密度梯度离心法富积循环肿瘤细胞,分析其与临床特征及化疗疗效的关系.结果 肺Ca组循环肿瘤细胞阳性率达72.00％,而COPD组阳性率为0,差异具有统计学意义(P<0.05).循环肿瘤细胞阳性表达与NSCLC病理类型、患者性别、年龄、吸烟情况等均无明显关系(P>0.05),仅与癌症分期相关(P<0.05).2个周期的化疗结束,发现对循环肿瘤细胞数目的评价与实体肿瘤疗效标准评价大致吻合,差异不具备统计学意义(P>0.05).相关分析发现这两种评价方法 呈负相关关系(γ=-0.465 P=0.021).结论 循环肿瘤细胞的存在与非小细胞肺癌的临床分期及化疗疗效有密切关系,可以用来预测非小细胞肺癌的转归.     

培美曲塞联合奥沙利铂治疗进展期肺腺癌的临床观察

目的 观察培美曲塞联合奥沙利铂治疗进展期肺腺癌的疗效及毒副反应.方法 51例经病理及组织细胞学证实为肺腺癌患者,既往一线化疗失败,接受培美曲塞联合奥沙利铂方案治疗,每例患者至少完成2周期.结果 51例患者中,CR 0例,PR 6例,SD 32例,PD 13例,有效率为11.76％ (6/51),疾病控制率为74.51％ (38/51).主要毒副反应为骨髓抑制、消化道反应、神经毒性.结论 培美曲塞联合奥沙利铂治疗进展期肺腺癌疗效较好,毒副反应可耐受.     

Pemetrexed in Previously Treated Non-small Cell Lung Cancer Patients with Poor Performance Status

Background and objective Pemetrexed have been approved for the treatment of patients affected by advanced non-small cell lung cancner(NSCLC) in progression after first-line chemotherapy.We evaluated the activity and feasibility of pemetrexed in previously treated NSCLC.Methods Patients with histologically or cytologically confirmed NSCLC were evaluated from April 2007 to March 2009.The patients had relapsed or progressed after prior chemotherapy treatment.Pemetrexed(500 mg/m2) was administered intravenously once every 3 weeks after progression to prior chemotherapy.The tumor response was evaluated according to RECIST criteria by chest CT at every 2 cycles of chemotherapy.Results A total 61 patients were eligible for analysis.Performance status of them(100%) was over 2.The response rate and disease control rate were 14.7% and 37.7% respectively.Non-squamous cell carcinoma histology was significantly associated with a superior response rate(P=0.045) and disease control rate(P=0.008).The median survival time and the median progression free survival(PFS) time were 6.11 months and 2.17 months,respectively.Comparing the efficacy of pemetrexed in these two settings [second-line versus(12/61) more than third(49/61)],there was no significant difference in regard to median survival(11.18 months vs 11.46 months,P=0.922,5),but PFS was more longer in third-or further-line groups than second-line group(1.39 months vs 2.25 months,P=0.015,3).Conclusion Pemetrexed is a feasible regimen in previously treated NSCLC with poor performance status.     

Autologous Tumor-specific Cytotoxic T Lymphocytes in a Patient with Lung Adenocarcinoma: Implications of the Shared Antigens Expressed in HLA-A24 Lung Cancer Cells

Human lung adenocarcinoma-specific cytotoxic T lymphocytes (CTL) were generated by multiple stimulations with autologous tumor cells (named A110L) from regional lymph node lymphocytes and tumor-infiltrating lymphocytes expanded by solid-phase anti-CD3 monoclonal antibody (mAb) and recombinant interleukin-2. The CTL lysed A110L but failed to kill either autologous B lymphocytes immortalized by the Epstein-Barr virus or K562. The killing activity of the CTL against autologous A110L was inhibited by anti-MHC class I mAb (W6/32), but not by anti-MHC class II mAb. The CTL produced interferon-γ and GM-CSF in response to A110L and the production was completely blocked by the addition of anti-MHC class I mAb. The HLA type of the CTL was HLA-A2/A24, B52/B54, Cw1/-. Allele-specific deletion of HLA-A2 molecules was observed in A110L by staining with anti-HLA-A2 mAb. A partial blocking effect on the cytokine production from the CTL was also obtained with anti-CD8, and anti-HLA-A24 mAbs, but not with anti-MHC class II, anti-CD4 and anti-HLA-A2 mAbs. To analyze further the mechanism of antigen recognition by the CTL, the cross reactivity of the CTL against several HLA-A locus-matched (HLA-A24+) and mismatched allogeneic tumor cells (HLA-A24-) was investigated. The A110L-specific CTL showed a weak but significant cytotoxicity against some HLA-A24 positive lung cancer cell lines, such as Sq-1 (HLA-A11/A24, squamous cell carcinoma) and PC-9 (HLA-A2/A24, adenocarcinoma), but failed to kill HLA-A locus-mismatched allogeneic tumors. This cross reactivity of the CTL against Sq-1 and PC-9 was blocked by anti-MHC class I mAb. These results thus demonstrate that shared common tumor antigens might exist among lung cancer cells in the context of HLA-A24.