OBJECTIVETo develop a novel rapid protocol for the detection of occult disseminated tumor cells by a recombinant herpes simplex virus expressing GFP (HSV(GFP)).

METHODSTumor cells of seven cell lines were exposed to HSV(GFP) and then examined for GFP expression by fluorescence microscopy. Various numbers of tumor cells (10, 100, 1000, 10 000) were mixed into 2 ml human whole blood, separated with lymphocytes separation medium, exposed to HSV(GFP), incubated at 37°C for 6 - 24 h and then counted for the number of green cells under the fluorescence microscope. Some clinical samples including peripheral blood, pleural effusion, ascites, spinal fluid from tumor-bearing patients were screened using this protocol in parallel with routine cytological examination.

RESULTSHSV(GFP) was able to infect all 7 tumor cell lines indicating that the HSV(GFP) can be used to detect different types of tumor cells. The detection sensitivity was 10 cancer cells in 2 ml whole blood. In the clinical samples, there were 4/15 positive by routine cytological examination but 11/15 positive by HSV(GFP), indicating a higher sensitivity of this new protocol.

CONCLUSIONRecombinant herpes simplex virus-mediated green fluorescence is a simple and sensitive technique for the identification of occult disseminated cancer cells including circulating tumor cells (CTCs).

Animals ; Cell Line, Tumor ; Cercopithecus aethiops ; Green Fluorescent Proteins ; metabolism ; Humans ; Neoplastic Cells, Circulating ; metabolism ; pathology ; Recombinant Proteins ; metabolism ; Sensitivity and Specificity ; Simplexvirus ; metabolism ; Vero Cells

OBJECTIVETo investigate the correlations between circulating tumor cell (CTC) and clinicopathologic characteristics of tumors obtained by core needle biopsy in axillary lymph node positive primary breast cancer patients.

METHODSThe peripheral venous blood samples were collected from 126 patients with axillary lymph node positive primary breast cancer and were detected to found CTCs using the CellSearch automatic detection system. The associations between CTCs and clinicopathologic characteristics of tumors were analyzed in axillary lymph node positive primary breast cancer patients. All patients were female, age ranging from 27 to 65 years (median, 49 years).

RESULTSOne or more CTCs were detected from the peripheral blood in 25.4% (32/126) patients. The positive rate of CTCs was 36.2% (17/47) in the human epidermal growth factor receptor 2 (HER-2) (+) patients, 19.0% (15/79) in the HER-2 (-) patients. In univariate analysis, there were significant differences about the positive rate of CTCs between the two groups (χ² = 4.592, P < 0.05). In multivariate analysis, the risk of circulating tumor cells positive in HER-2 (+) patients was 2.712 times higher than in HER-2 (-) patients (OR = 2.712, 95% CI: 1.117-6.584, P = 0.027), whereas the positive rate of CTCs in axillary lymph node positive primary breast cancer patients showed no significant differences among the different subgroups with regards to age, menopausal status, the T staging of the tumor, histological type, histological grade, hormone receptor status and Ki-67 expression level (P > 0.05).

CONCLUSIONSThere are significant correlations between the presence of CTCs and the HER-2 status of the tumor in axillary lymph node positive primary breast cancer patients. No significant correlations are found between the presence of CTCs and the age, menopausal status, T staging of the tumor, histological type, histological grade, hormone receptor status and Ki-67 expression level.

Adult ; Aged ; Axilla ; pathology ; Breast Neoplasms ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Receptor, ErbB-2 ; metabolism

Carcinoma, Renal Cell ; immunology ; pathology ; surgery ; Humans ; Keratin-19 ; metabolism ; Kidney Neoplasms ; immunology ; pathology ; surgery ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Nephrectomy ; Vena Cava, Inferior ; pathology ; surgery

OBJECTIVETo explore whether cancer cells abide by the mechanism of epithelial-mesenchymal transition (EMT) in the process of invasion and metastasis by comparing histology and protein expression of E-cadherin and vimentin among primary, metastatic carcinomas and their emboli.

METHODSA total of 68 tissue specimens in 59 cases of primary adenocarcinoma or squamous cell carcinoma and their lymphatic metastasis were collected, of which there were 13 well differentiated, 11 moderately differentiated, 30 poorly differentiated tumors and 14 lymphatic metastases. The morphology and the expression of E-cadherin and vimentin proteins were assessed by H-E stain and immunohistochemistry.

RESULTSThe overall morphology of the primary cancers and their tumor emboli was similar. Among 54 primary cancers, 50 cases were positive for E-cadherin and 22 cases were positive for vimentin. Fifty-one cases were positive for E-cadherin and 22 cases were positive for vimentin in the tumor emboli, with no statistical difference (P = 0.804, P = 0.842). Among 14 cases of lymphatic metastasis, 12 cases were positive for E-cadherin and 6 cases were positive for vimentin, and the tumor emboli in 12 cases were positive for E-cadherin and 7 cases were positive for vimentin, with statistical difference (P = 0.084, P = 0.878). There were no significant difference of E-cadherin and vimentin protein expression between the cancer tissue and its emboli (P = 0.410, P = 0.824). A subset of tumor cells in cancer emboli expressed E-cadherin at a high level without vimentin expression, whereas other cells in tumor emboli showed an opposite expression pattern.

CONCLUSIONSThere is no significant difference of EMT characteristics among primary cancer, lymphatic metastases and their cancer emboli. Cancer thrombus contains both EMT and non-EMT cells. Further studies are required to elucidate the role of EMT in the processes of tumor invasion and metastasis.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Cadherins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Epithelial-Mesenchymal Transition ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasms ; metabolism ; pathology ; Neoplastic Cells, Circulating ; metabolism ; pathology ; Vimentin ; metabolism

We report here a case of a 59-yr-old man with CD4+ T-cell large granular lymphocytic leukemia (T-LGL). Peripheral blood examination indicated leukocytosis (45x10(9) cells/L) that consisted of 34% neoplastic lymphoid cells. Other laboratory results indicated no specific abnormalities except for serum antinuclear antibody titer (1:640), glucose (1.39 g/L), and hemoglobin A1c (7.7%) levels. Computed tomography indicated multiple small enlarged lymph nodes (<1 cm in diameter) in both the axillary and inguinal areas, a cutaneous nodule (1.5 cm in diameter) in the left suboccipital area, and mild hepatosplenomegaly. Bone marrow examination revealed hypercellular marrow that consisted of 2.4% neoplastic lymphoid cells. The neoplastic lymphoid cells exhibited a medium size, irregularly shaped nuclei, a moderate amount of cytoplasm, and large granules in the cytoplasm. Immunohistochemical analysis indicated CD3+, CD4+, T-cell receptor betaF1+, granzyme B+, and TIA1+. Flow cytometric analysis of the neoplastic lymphoid cells revealed CD3+, cytoplasmic CD3+, CD4+, and CD7+. Cytogenetic analysis indicated an abnormal karyotype of 46,XY,inv(3)(p21q27),t(12;17)(q24.1;q21),del(13)(q14q22)[2]/46,XY[28]. The patient was diagnosed with CD4+ T-LGL and received chemotherapy (10.0 mg methotrexate). This is the second case of CD4+ T-LGL that has been reported in Korea.
Antibodies, Antinuclear/analysis ; Blood Glucose/analysis ; Bone Marrow Cells/metabolism/pathology ; Hemoglobin A, Glycosylated/metabolism ; Humans ; Immunohistochemistry ; Immunophenotyping ; Karyotyping ; Leukemia, Large Granular Lymphocytic/*diagnosis/pathology/radiography ; Lymph Nodes/pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating/metabolism/pathology ; Tomography, X-Ray Computed

We report here a case of a 59-yr-old man with CD4+ T-cell large granular lymphocytic leukemia (T-LGL). Peripheral blood examination indicated leukocytosis (45x10(9) cells/L) that consisted of 34% neoplastic lymphoid cells. Other laboratory results indicated no specific abnormalities except for serum antinuclear antibody titer (1:640), glucose (1.39 g/L), and hemoglobin A1c (7.7%) levels. Computed tomography indicated multiple small enlarged lymph nodes (<1 cm in diameter) in both the axillary and inguinal areas, a cutaneous nodule (1.5 cm in diameter) in the left suboccipital area, and mild hepatosplenomegaly. Bone marrow examination revealed hypercellular marrow that consisted of 2.4% neoplastic lymphoid cells. The neoplastic lymphoid cells exhibited a medium size, irregularly shaped nuclei, a moderate amount of cytoplasm, and large granules in the cytoplasm. Immunohistochemical analysis indicated CD3+, CD4+, T-cell receptor betaF1+, granzyme B+, and TIA1+. Flow cytometric analysis of the neoplastic lymphoid cells revealed CD3+, cytoplasmic CD3+, CD4+, and CD7+. Cytogenetic analysis indicated an abnormal karyotype of 46,XY,inv(3)(p21q27),t(12;17)(q24.1;q21),del(13)(q14q22)[2]/46,XY[28]. The patient was diagnosed with CD4+ T-LGL and received chemotherapy (10.0 mg methotrexate). This is the second case of CD4+ T-LGL that has been reported in Korea.
Antibodies, Antinuclear/analysis ; Blood Glucose/analysis ; Bone Marrow Cells/metabolism/pathology ; Hemoglobin A, Glycosylated/metabolism ; Humans ; Immunohistochemistry ; Immunophenotyping ; Karyotyping ; Leukemia, Large Granular Lymphocytic/*diagnosis/pathology/radiography ; Lymph Nodes/pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating/metabolism/pathology ; Tomography, X-Ray Computed

OBJECTIVETo explore the detection efficiency of circulating tumor cells (CTCs) in patients with hepatocellular carcinoma (HCC).

METHODSImmunomagnetic negative enrichment by nanometer magnetic beads and label-free capture with Captor(TM) system were used to isolate and enrich CTCs from peripheral blood of HCC patients, and epithelial and HCC markers were applied to identify CTCs by immunofluorescence staining. CTCs were detected in 50 HCC patients before and after hepatectomy to test the method for isolation, enrichment and identification. The dynamic changes of pre- and post-operative CTCs' numbers were compared. The clinical data were analyzed using SPSS 19.0 software.

RESULTSNegative enrichment methods by nanometer magnetic beads and label-free capture using Captor(TM) system were both suitable for CTCs isolation and enrichment in HCC patients. The positive detection rate of CTCs in HCC patients via negative enrichment was 96.0% (48/50), the preoperative median number of CTCs was 16 per 7.5 ml blood, and the postoperative median number was 17 per 7.5 ml blood.

CONCLUSIONSBoth negative enrichment and Captor(TM) system are suitable for isolation and enrichment of CTCs in HCC patients. There is a significant difference in the numbers of CTCs before and after operation, and dynamic detection of CTCs will provide helpful prognostic information for HCC patients in clinics.

Adult ; Aged ; Antigens, Neoplasm ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Adhesion Molecules ; metabolism ; Epithelial Cell Adhesion Molecule ; Female ; Hepatectomy ; Humans ; Immunomagnetic Separation ; methods ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; metabolism ; pathology

AC133 Antigen ; Animals ; Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antineoplastic Agents ; pharmacology ; Biomarkers, Tumor ; metabolism ; Glycoproteins ; metabolism ; Humans ; Neoplasms ; drug therapy ; metabolism ; pathology ; Neoplastic Cells, Circulating ; pathology ; Neoplastic Stem Cells ; drug effects ; metabolism ; pathology ; Peptides ; metabolism ; Receptors, Notch ; metabolism ; Signal Transduction ; Stem Cells ; drug effects ; metabolism ; pathology

OBJECTIVETo establish a quantitative method to detect circulating tumor cells (CTC) in patients with small cell lung cancer, and analyze its sensitivity and stability.

METHODSA specific primer and probe for prepro-gastrin-releasing peptide (preproGRP) was designed and a quantitative RT-PCR method was established to detect preproGRP mRNA. Cell incorporation method was used to evaluate the sensitivity. Magnetic cell sorting (MACS) was used to isolate and purify CTC from peripheral blood, and the MACS in combination with morphological diagnosis were used for cell counting.

RESULTSThe isolation rate of CTC by MACS was 30% and the lower detection limit was 5 cells per ml blood. The sensitivity of quantitative RT-PCR in detection of preproGRP mRNA in CTC was 0.64 cells per reaction, and the lower detection limit was 50 cells per ml blood, which was lower than that of MACS. However, the cell numbers calculated by Ct value was in greater accordance (about 80%) with actual cell numbers than that obtained by MACS.

CONCLUSIONSPreproGRP quantitative RT-PCR and MACS have both advantages and disadvantages in detecting CTC of SCLC patients. MACS has a higher sensitivity, and is more favorable when CTC count is below 50 per ml blood. Meanwhile, preproGRP mRNA quantitative RT-PCR is more reliable in calculating actual cell numbers.

Humans ; Immunomagnetic Separation ; Lung Neoplasms ; blood ; metabolism ; pathology ; Neoplastic Cells, Circulating ; Peptides ; genetics ; metabolism ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Small Cell Lung Carcinoma ; blood ; metabolism ; pathology

OBJECTIVETo study the clinical significance of detecting p53 gene mutation expression in colorectal cancer cells of peripheral blood.

METHODSFlow cytometry (FCM) was used to detect p53 gene mutation expression in peripheral blood cancer cells of 128 patients with colorectal cancer. Experimental data were analyzed by SPSS (v.11.0) software.

RESULTSThe lymph node metastasis showed the significant difference statistically (P<0.01) between p53 positive and negative expression in the colorectal cancer patients. The mutation p53 expression associated with existing histological differentiation (r=0.8476, P<0.05). A lymph node metastasis difference was observed between left and right colorectal cancers of mutation p53 positive expression.

CONCLUSIONDetecting the mutation p53 expression in cancer cells of peripheral blood might be helpful to the early diagnosis of colorectal cancer.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; diagnosis ; genetics ; pathology ; DNA, Neoplasm ; analysis ; Female ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Genes, p53 ; genetics ; Humans ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; metabolism ; Tumor Suppressor Protein p53 ; genetics