Clinical Significance of Circulating Tumor Cells and Lymph Node Micrometastases in Gastrointestinal cancer

(1) CellSearch System检测胃癌患者血液中循环肿瘤细胞本研究的目的是应用CellSearch System （CSS）评价胃癌患者外周血（PB）和门静脉血（PV）的CTCs，并探讨其与复发的关系。【方法】50例T1-3型胃癌患者，在胃切除术前取PB 10ml，术中取PV 10ml。【结果】PB中ctc含量为18%，PV中ctc含量为24%，PB和/或PV中ctc含量为28%。T1组CTCs占8%，T2组26%，T3组44%。CTCs检出率与肿瘤分期（p=0.007）、血管浸润（p=0.008）、淋巴浸润（p=0.007）、淋巴结转移（p=0.01）相关。11例（78.5%）CTCs患者出现复发，有无CTCs患者的总生存率差异有统计学意义（p=0.0049）。【结论】ctc的鉴别对胃癌术后随访及辅助化疗具有重要意义。(2)-1利用多重实时RT-PCR检测快速诊断胃癌淋巴结转移的效用本研究的目的是评估在SmartCycler上运行的原型RT-PCR检测的效用[j]，可在40分钟内快速诊断胃癌淋巴结转移。本研究的目的是验证SmartCycler的效用。R]与使用LightCycler的传统RT-PCR相比^[〇！R]。【患者与方法】选取8例晚期胃癌患者的47个明显转移淋巴结和22例非恶性肿瘤患者的良性淋巴结，均经知情同意。我们用RT-PCR检测了转移性淋巴结，使用了多个标记，使用了CEA和LightCycler的CK19。和SmartCycler^[〇！R]。【结果】在LightCycler中，CEA和CK19的敏感性分别为91.5%和70.2%。R]，在原型分析系统中分别为97.9%和95.7%。在多重检测中，LightCycler^[〇！]的灵敏度为91.5%。系统和97.9%的SmartCycler^[〇！]R),分别。【结论】SmartCycler的RT-PCR快速诊断淋巴结转移[R]与传统的LightCycler一样具有较高的淋巴结转移检测精度。在这项研究中，SmartCycler^[〇][R]对胃癌淋巴结转移的诊断有利。(2)-2 GeneSearch BLN检测在快速评估乳腺癌前哨淋巴结中的应用我们的目标是通过将乳腺淋巴结（BLN）检测与常规病理检查进行比较，评估该检测的临床应用，这是一种实时RT-PCR检测SLN转移的方法。【方法】117例乳腺癌患者行乳腺手术并行SLN活检。每个SLN沿着最长维度的平面切成两半。每个淋巴结的一半用BLN试验的两种标记物，mammaglobin （MG）和cytokeratin 19 （CK19）检测，另一半用苏木精和伊红染色（HE）和免疫组织化学染色（IHC）检测胰腺细胞角蛋白。【结果】117例患者共获得sln 204枚。HE检出31例sln转移（15.2%），IHC检出6例sln转移；32例患者40例sln经BLN检测转移阳性。检测结果与HE、IHC病理诊断相关（p<0.001）。与TNM分类的病理结果相比，BLN检测对大转移瘤的敏感性为95.7%，对微转移瘤的敏感性为60.0%，对分离肿瘤细胞的敏感性为55.6%。【结论】2标记物BLN检测性能可与常规病理检查相媲美，且分析组织的数量多于常规病理检查。因此，临床术中使用BLN检测sln可能会减少腋窝淋巴结清扫的第二次手术。少

(1) Circulating tumor cells in blood detected by CellSearch System in gastric cancer The purposes of this study are to evaluate CTCs of peripheral blood (PB) and portal vein blood (PV) with gastric cancer patients using CellSearch System (CSS), and to investigate the relation of the recurrence. 【Methods】 In 50 patients with T1-3 gastric cancer, PB was collected 10ml before the gastrectomy, and PV was collected 10ml during operation. 【Results】CTCs were detected in 18% from PB, 24% from PV and 28% from PB and/or PV. CTCs were found in 8% with T1, 26% with T2, 44% with T3. Detection rate of CTCs were correlated with the results of Stage (p=0.007), vessel invasion (p=0.008), lymphatic invasion (p=0.007) and lymph node metastasis (p=0.01). The recurrences were detected in 11 patients (78.5%) with CTCs, and there was statistically significant difference in over all survival between patients with or without CTCs (p=0.0049). 【Conclusions】The identification of CTCs is very useful to follow up a … More fter operation and to perform the adjuvant chemotherapy in gastric cancer.(2)-1 The utility of rapid diagnosis of lymph node metastasis in gastric cancer using a multiplex real-time RT-PCR assayThe purpose of this study was to evaluate the utility of a prototype RT-PCR assay run on the SmartCycler^[○!R] that enables the rapid diagnosis of lymph node metastasis in gastric cancer within 40 minutes. The purpose of this study was to verify the utility of the SmartCycler^[○!R] as compared to conventional RT-PCR using the LightCycler^[○!R]. 【Patients and methods】47 overt metastatic lymph nodes from 8 patients with advanced gastric cancer and 22 benign lymph nodes from patients without malignant tumor who received surgery were obtained with informed consent. We examined the metastatic lymph nodes by RT-PCR using multiple markers using CEA and CK19 by the LightCycler^[○!R] and SmartCycler^[○!R]. 【Results】 In the singlex assay, the sensitivity of CEA and CK19 was 91.5% and 70.2% in the LightCycler^[○!R], and 97.9% and 95.7% in the prototype assay system, respectively. In the multiplex assay, the sensitivity was 91.5% in the LightCycler^[○!R] system and 97.9% in the SmartCycler^[○!R], respectively. 【Conclusion】Rapid diagnosis for lymph node metastasis using RT-PCR by the SmartCycler^[○!R] has equally high precision for detecting lymph node metastasis as the conventional LightCycler^[○!R]. In this study the SmartCycler^[○!R] is advantageous for the diagnosis of lymph node metastasis in gastric cancer when run with the prototype assay.(2)-2 Utility of the GeneSearch BLN Assay for Rapid Evaluation of Sentinel Lymph Nodes in Breast CancerOur goal was to evaluate the clinical application of the Breast Lymph Node (BLN) assay, a real-time RT-PCR assay for SLN metastases, by comparing this test with routine pathological examination. 【Methods】One hundred seventeen patients with breast cancer underwent breast surgery with SLN biopsy. Each SLN was cut in half along the plane of the longest dimension. Half of each node was examined by the BLN assay's 2 markers, mammaglobin (MG) and cytokeratin 19 (CK19), and the other half was examined by hematoxylin and eosin staining (HE) and immunohistochemical staining (IHC) for pancytokeratins. 【Results】A total of 204 SLNs were obtained from 117 patients. HE identified metastases in 31 SLNs (15.2%), and IHC detected metastases in 6 SLNs ; 40 SLNs from 32 patients were positive for metastasis by the BLN assay. The assay results correlated with the pathological diagnoses by HE and IHC (p<0.001). BLN Assay sensitivity compared with pathological findings classified according to TNM was 95.7% for macrometastases, 60.0% for micrometastases, and 55.6% for isolated tumor cells. 【Conclusion】The 2-marker BLN assay has performance comparable to, and analyzes more tissue than, routine pathological examination. Therefore, clinical intraoperative use of the BLN assay for SLNs may result in reduction of the second surgery for axillary lymph node dissections. Less