mRNA-positive Cells In Peripheral Blood Research Articles

CK-19 mRNA-positive cells in peripheral blood predict treatment efficacy and survival in small-cell lung cancer patients

Small-cell lung caner (SCLC) is the most aggressive form of lung cancer. The aim of this study was to investigate whether the presence of cytokeratin-19 (CK-19) mRNA-positive circulating tumor cells (CTCs) predicts treatment response, progression-free survival (PFS), and overall survival (OS) in SCLC patients who received standard therapy. Fifty-five SCLC patients were enrolled in this single-center prospective study. CK-19 mRNA-positive CTCs in blood samples were detected using real-time quantitative-PCR assay before the initiation of chemotherapy (B0) and after one chemotherapy cycle (B1) and three chemotherapy cycles (B3). The association with known prognostic factors and the effect of CK-19 mRNA-positive CTCs on patients' prognosis were analyzed. Patients with positivity for CK-19 mRNA-positive CTCs at B0, B1, and B3 time points had shorter PFS and OS compared with patients without (P=0.014 and P=0.01, respectively, at B0; P=0.008 and P=0.002, respectively, at B1; P=0.003 and P=0.001, respectively, at B3). Conversion of initial positivity for CK-19 mRNA-positive CTCs to negativity at B1 and B3 time points was associated with longer PFS and OS compared with patients with persistent positivity at three time points (P=0.008 and P=0.010, respectively). Multivariate analysis demonstrated that the presence of CK-19 mRNA-positive CTCs at B0, B1, and B3 time points remained strong predictors of PFS and OS after adjustment for clinically significant factors. In conclusion, detection of CK-19 mRNA-positive CTCs before and during chemotherapy is an accurate indication of subsequent disease progression and mortality for SCLC patients.

MUC2 mRNA detection in peripheral blood and bone marrow of breast cancer patients reveals micrometastasis

Tumor dissemination to distant organ is the main cause of death. Therefore there is urgent need to set up sensitive methods for early detection of circulating tumor cells (CTCs) and disseminated tumor cells (DTCs) in peripheral blood (PB) and bone marrow (BM) specimens of breast cancer patients. We aim to detect MUC2 mRNA positive cells in PB and BM of breast cancer patients; to relate this to patient relapse. In this study to detect MUC2 mRNA positive cells (tumor marker), PB and BM samples were collected from 50 breast cancer patients after operation and before adjuvant therapy with 20 PB from healthy individuals as negative controls. Chi-square test was used to analyze data. MUC2 mRNA by using Real-time PCR was detected in 9 (18%) of PB and in 10 (20%) of BM samples and none of the healthy individuals. The relapse rate among MUC2-positive patients was significance in BM (P

1092 Detection of cytokeratin-19 mRNA-positive cells in peripheral blood and bone marrow of patients with operable breast cancer

1092 Detection of cytokeratin-19 mRNA-positive cells in peripheral blood and bone marrow of patients with operable breast cancer

Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method

A real-time reverse transcriptase-polymerase chain reaction (RT-PCR) method for detection of cytokeratin 20-positive cells in blood characterized by two novel features was developed and tested on 99 patients with colorectal cancer, 110 with breast cancer, and 150 healthy subjects. To optimize the specificity and sensitivity of the method, two novel features were used. First, a primer overlapping two adjacent exons was generated to inhibit nonspecific amplification both in healthy donors and cancer patients; second, a non-end-point first-round amplification was used to increase sensitivity. The number of first-round cycles was chosen to reach the highest level of sensitivity while conserving quantitative characteristics. PCR efficiency increased from 88.9% in single-round RT-PCR to 99.0% in nested real-time RT-PCR. To establish sensitivity and specificity of the method, HT29 cells were serially diluted with normal blood. Detection limit improved from 100 HT29 cells (single-round RT-PCR) to 1 to 10 cells (nested real-time RT-PCR) per 3 ml of whole blood. None of the healthy subjects was positive, whereas 22 and 29% of all colorectal and breast cancer patients, respectively, had cytokeratin 20 cell equivalents in blood. The association between cytokeratin 20 cell equivalents and metastasis was statistically significant for breast (P = 0.026) but not colorectal cancer patients (P = 0.361). Negativity of all 150 healthy controls examined confers diagnostic potential to the method.

PD-018 Real-time quantification of CK-19 mRNA-positive cells inperipheral blood of patients using with non-small cell lung cancer (NSCLC) using Real Time RT-PCR

PD-018 Real-time quantification of CK-19 mRNA-positive cells inperipheral blood of patients using with non-small cell lung cancer (NSCLC) using Real Time RT-PCR