ABSTRACT: ObjectiveThe purpose of this study is to explore RT-PCR method to set up the examination platform for detecting circulating tumor cells
(CTC) in peripheral blood from metastatic breast cancer patients. The primary endpoint is to find out the correlation of existence
of CTC with clinical responses and progression-free survival (PFS).
MethodsThe breast cancer cell line MCF-7 was serially diluted into the peripheral blood from 45 healthy donors to set up the sensitivity
of RT-PCR assay. The expression of CK19 mRNA was amplified from both 49 patients and 45 healthy donors respectively. The CK19
protein quantity from plasma was measured by competitive inhibition ELISA assay.
ResultsThe sensitivity of RT-PCR could reach 1/106–107 white blood cells with specificity of 95.6%. The objective response rate (ORR) of patients with CK19 mRNA-negative undertaken
one cycle chemotherapy was significantly higher than those with positive (P<0.0001). PFS among CK19 mRNA-negative patients was also increased, although there was no significance (P=0.098). The results of ELISA assay showed that CK19 protein was decreased significantly after one cycle chemotherapy, which
gave rise to a little higher ORR (P=0.015) and increased PFS (P=0.016).
ConclusionPatients with unamplified CK19 mRNA after one cycle chemotherapy could achieve better radiographic evaluation and increased
PFS, which was showed to be of consistency with the CK19 protein assay among the patients treated.
Key wordsBreast cancer-Circulating tumor cells-CK19-RT-PCR-ELISA
Chinese Journal of Cancer Research 04/2012; 22(3):201-210. · 0.18 Impact Factor
ABSTRACT: To test for circulating tumor cells (CTCs) relying on epithelial cellular adhesion molecule (EpCAM) expression in metastatic breast cancer by quantitative real-time reverse transcription-PCR.
In the study,47 metastatic breast cancer patients were evaluated by quantitative real-time PCR for detecting EpCAM mRNA. In addition, analyses were carried out for their correlation with patients' clinicopathologic features, response, and the time to progression (TTP).
The sensitivity of EpCAM mRNA in the metastatic breast cancer patients was about 40%. However, the specificity of EpCAM mRNA for 20 healthy controls was 100%. TTP was calculated, and compared with that between EpCAM mRNA-positive and EpCAM mRNA-negative groups. For the retrospective study, the median TTP was 7.1 months and 11.1 months (P=0.013) for patients with EpCAM mRNA-positive and EpCAM mRNA-negative, respectively, after the first cycle chemotherapy. Moreover, a statistically significant correlation was demonstrated between EpCAM mRNA and TTP in patients who underwent the first or the second-line chemotherapy (P=0.018), but there was no significance in the patients pretreated with two or more previous chemotherapy lines (P=0.471).
This study provides evidence of the presence of EpCAM mRNA in approximately 40% of patients with metastatic breast cancer. There is a strong correlation between EpCAM mRNA results after the first cycle therapy and TTP in metastatic breast cancer patients, and EpCAM mRNA positive after chemotherapy may predict shorter TTP.
Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences 04/2012; 44(2):275-80.
ABSTRACT: Chemotherapy plays an important role in the treatment of metastatic breast cancer. It is important to monitor chemotherapeutic efficacy, to find a simple and efficient tool to guide treatment, and to predict the efficacy of treatment in a timely and accurate manner. This study aimed to detect mucin-1 (MUC1)-positive circulating tumor cells and MUC1 protein in the peripheral blood of patients with metastatic breast cancer and to investigate their relationship to chemotherapeutic efficacy. MUC1 mRNA was detected in the peripheral blood of 34 patients with newly diagnosed metastatic breast cancer by reverse transcription-polymerase chain reaction. The positive rates of MUC1 mRNA were 88.2% before chemotherapy and 70.6% after chemotherapy, without a significant difference (P=0.564); MUC1 mRNA expression before chemotherapy had no correlation with treatment effectiveness (P=0.281). The response rate of MUC1 mRNA-negative patients after first-cycle chemotherapy was significantly higher (P=0.009) and the progression-free survival (PFS) was clearly longer than those of MUC1 mRNA-positive patients (P=0.095). MUC1 protein in peripheral blood plasma was detected by an ELISA competitive inhibition assay. The patients with decreased MUC1 protein after chemotherapy had a significantly longer PFS than those with elevated MUC1 protein (P=0.044). These results indicate that the outcomes of MUC1 mRNA-negative patients after chemotherapy are better than those of MUC1 mRNA-positive patients. In addition, patients with decreased expression of MUC1 protein have a better PFS.
Chinese journal of cancer 01/2011; 30(1):54-61.
ABSTRACT: To investigate apoptosis in human pancreatic cancer cells induced by Triptolide (TL), and the relationship between this apoptosis and expression of caspase-3' bcl-2 and bax.
Human pancreatic cancer cell line SW1990 was cultured in DMEM media for this study. MTT assay was used to determine the cell growth inhibitory rate in vitro. Flow cytometry and TUNEL assay were used to detect the apoptosis of human pancreatic cancer cells before and after TL treatment. RT-PCR was used to detect the expression of apoptosis-associated gene caspase-3' bcl-2 and bax.
TL inhibited the growth of human pancreatic cancer cells in a dose-and time-dependent manner. TL induced human pancreatic cancer cells to undergo apoptosis with typically apoptotic characteristics. TUNEL assay showed that after the treatment of human pancreatic cancer cells with 40 ng/mL TL for 12 h and 24 h, the apoptotic rates of human pancreatic cancer cells increased significantly. RT-PCR demonstrated that caspase-3 and bax were significantly up-regulated in SW1990 cells treated with TL while bcl-2 mRNA was not.
TL is able to induce the apoptosis in human pancreatic cancer cells. This apoptosis may be mediated by up-regulating the expression of apoptosis-associated caspase-3 and bax gene.
World Journal of Gastroenterology 04/2008; 14(10):1504-9. · 2.47 Impact Factor