Effects of RNAi-mediated matrix metalloproteinase-2 gene silencing on the invasiveness and adhesion of esophageal carcinoma cells, KYSE150.

BACKGROUND: Esophageal carcinoma is one of the main malignancies in China. Previous studies indicated that matrix metalloproteinases (MMPs) play important roles in the process of tumor invasion and metastasis in several types of solid tumors. Among all of the MMPs, MMP-2 is one of the MMPs closely associated with tumor invasion. In this study, we suppressed MMP-2 expression with RNA interference and then observed inhibitory effects on the invasion and migration of the esophageal carcinoma cell line KYSE150. METHODS: Three target sequences were selected and siRNA against MMP-2 mRNA were synthesized. After being transfected by the transfection complexes, the MMP-2 expression of KYSE150 cells, which overexpresses MMP-2, were examined by Western blot analysis and real-time polymerase chain reaction (PCR). Cell migration and invasion were measured with migration assay and Boyden chamber assays, respectively. RESULTS: RNAi against MMP-2 successfully inhibited the mRNA and protein expression of MMP-2 in the esophageal carcinoma cell line KYSE150. MMP-2 knockdown inhibited the invasion and migration of esophageal carcinoma cell line KYSE150. CONCLUSIONS: These findings suggested that the RNAi approach towards MMP-2 may be a potentially effective therapeutic method for the treatment of esophageal carcinoma.

Neoadjuvant chemoradiotherapy could improve survival outcomes for esophageal carcinoma: a meta-analysis.

BACKGROUND: The effectiveness of neoadjuvant chemoradiotherapy in patients with resectable esophageal carcinoma remains controversial. AIMS: The purpose of this study was to assess the effect of neoadjuvant chemoradiotherapy on operable esophageal carcinoma. METHODS: We searched PubMed, EMBASE and Web of Science and identified all randomized controlled trials published up until July 2011 that directly compared chemoradiotherapy followed by surgery with surgery alone. The risk ratio (RR) with its corresponding 95 % confidence interval (CI) was the principal measure of effects. RESULTS: Twelve randomized controlled trials that met our inclusion criteria were identified. Chemoradiotherapy followed by surgery was associated with significantly improved 1-year (RR = 0.86, 95 % CI = 0.74-0.98, P = 0.03), 3-year (RR = 0.82, 95 % CI = 0.73-0.92, P = 0.0007) and 5-year (RR = 0.83, 95 % CI = 0.72-0.96, P = 0.01) survival times compared with surgery alone. Subgroup analysis suggested that this benefit was associated with concurrent chemoradiotherapy but not sequential chemoradiotherapy. Neoadjuvant chemoradiotherapy could improve 3- and 5-year survival outcomes for squamous cell carcinoma but not those for adenocarcinoma. Postoperative morbidity (RR = 0.97, 95 % CI = 0.86-1.09, P = 0.56) and mortality (RR = 1.56, 95 % CI = 0.97-2.50, P = 0.07) did not increase in patients treated by chemoradiotherapy. CONCLUSIONS: Our findings revealed that compared with surgery alone, neoadjuvant chemoradiotherapy was associated with improved 1-, 3- and 5-year survival times, but not associated with increased postoperative morbidity and mortality in patients with esophageal carcinoma.

Nitrogen and titanium-codoped porous carbon nanocomposites derived from metal-organic framework as cathode to address polysulfides shuttle effects by Ti-assisted N-inhibiting strategy.

To address the problem of shutting effect of Li-S batteries, we used Ti-based MOF as precursor to obtain a conductive matrix with dual inhibitors. The target material, namely NTiPC, shown remarkable discharge capacity with 1178 mA h g-1, and maintained at 732 mA h g-1 after 100 cycles. The results indicated the N- and Ti-active sites synergistic acted with conductive framework can facilitate binding reaction between matrix and polysulfides.

Effects of RNA silencing of matrix metalloproteinase-2 on the growth of esophageal carcinoma cells in vivo.

Esophageal carcinoma is one of the most common malignancies in China. Previous studies reported that matrix metalloproteinases (MMPs) have important roles in the progression and invasion of numerous types of solid tumors. Among the MMPs, MMP-2 has been closely associated with tumor growth and invasion. In the present study, a short hairpin RNA (shRNA) lentiviral expression vector targeting the MMP-2 gene was constructed in order to observe the inhibitory effect of MMP-2 gene silencing on the growth of the KYSE150 esophageal carcinoma cell line in vivo. Three small hairpin RNA sequences targeting MMP-2 were designed and cloned into lentiviral vectors. Following transfection of the lentiviral vectors into KTSE150 cells, MMP-2 mRNA and protein expression levels were examined by reverse transcription-quantitative polymerase chain reaction and western blotting, and the growth rate of cells was analyzed by MTT assays. Subsequently, tumor growth was assessed in nude mice. Lentivirus-mediated RNA interference effectively inhibited the expression of MMP-2 mRNA and protein in KYSE150 esophageal carcinoma cells, and suppressed the growth of esophageal carcinoma cells in vivo. The results of the present study suggested that lentivirus-mediated gene therapy targeting MMP-2 may be an attractive strategy for the treatment of esophageal carcinoma and justifies the performance of further studies on the application of lentivirus vectors to cancer gene therapy.

Expression of surfactant protein-A in exhaled breath condensate of patients with chronic obstructive pulmonary disease.

Pulmonary surfactant protein A (SP-A) has been associated with host defense in the lung, and contributes to the pathogenesis of chronic obstructive pulmonary disease (COPD). The present study aimed to determine a non­invasive method of measurement of SP­A, and further examine the expression levels of SP­A in patients with COPD. SP­A was detected in the exhaled breath condensate (EBC) obtained from patients with COPD and from non­COPD subjects. The individuals recruited for the present study comprised 60 subjects with and without COPD, who underwent lobectomy for a solitary peripheral lung nodule. EBC was collected using a condenser, and an enzyme­linked immunosorbent assay (ELISA) was used to measure the levels of SP­A. Tissue samples were obtained during lobectomy through resection of the adjacent lung tissues, located >5 cm from the nodule. Western blot analysis and immunohistochemistry were used to measure SP­A and SP­A­positive type II pneumocytes. The results demonstrated that SP­A was detectable in the EBC of all subjects. The results of the ELISA and western blotting demonstrated that the expression levels of SP­A were significantly decreased in patients with COPD, compared with the non­COPD subjects. The reduction of SP­A­positive type II pneumocytes was associated with the expression levels of SP­A. Decreased expression levels of SP­A in EBC were associated with a higher degree of airway limitation. These results suggested that the measurement of SP­A levels in the EBC may serve as a method for monitoring airway obstruction in patients with COPD. Further investigations are required in order to examine these observations further and to elucidate the underlying mechanisms.

Circulating tumor DNA identified by targeted sequencing in advanced-stage non-small cell lung cancer patients.

Non-small cell lung cancers (NSCLC) have unique mutation patterns, and some of these mutations may be used to predict prognosis or guide patient treatment. Mutation profiling before and during treatment often requires repeated tumor biopsies, which is not always possible. Recently, cell-free, circulating tumor DNA (ctDNA) isolated from blood plasma has been shown to contain genetic mutations representative of those found in the primary tumor tissue DNA (tDNA), and these samples can readily be obtained using non-invasive techniques. However, there are still no standardized methods to identify mutations in ctDNA. In the current study, we used a targeted sequencing approach with a semi-conductor based next-generation sequencing (NGS) platform to identify gene mutations in matched tDNA and ctDNA samples from 42 advanced-stage NSCLC patients from China. We identified driver mutations in matched tDNA and ctDNA in EGFR, KRAS, PIK3CA, and TP53, with an overall concordance of 76%. In conclusion, targeted sequencing of plasma ctDNA may be a feasible option for clinical monitoring of NSCLC in the near future.

High Serum CEA and CYFRA21-1 Levels after a Two-Cycle Adjuvant Chemotherapy for NSCLC: Possible Poor Prognostic Factors.

OBJECTIVE: The aim of this study was to test whether carcinoembryonic antigen (CEA) and cytokeratin 19 fragments (CYFRA21-1) can be used as a prognostic factor for non-small-cell lung cancer (NSCLC) after two cycles of adjuvant chemotherapy in NSCLC patients. METHODS: A total of 169 patients underwent at least two cycles of adjuvant chemotherapy. The serum levels of CEA and CYFRA21-1 were recorded after the second cycle of chemotherapy, and the patient follow-up was conducted. Overall survival (OS) and disease-free survival (DFS) were used as the primary endpoint and the secondary endpoint, respectively. RESULTS: The high levels of CEA and CYFRA21-1 after two cycles of adjuvant chemotherapy were poor prognostic factors for OS, with risk ratios (RR) of 2.003 and 1.702, respectively. A high CEA level was a poor prognostic factor (RR 1.152) for DFS. The median survival time (MST) of the high CEA level group was 26 months, whereas that of the normal group was 61 months (P<0.0001). The median DFS time of the high CEA group and the normal group was 34 and 53 months, respectively (P<0.0001). The MST of the high CYFRA21-1 group and the normal group was 43 and 56 months, respectively (P<0.0001). CONCLUSIONS: The high serum levels of CEA or CYFRA21-1 after two cycles of adjuvant chemotherapy are poor prognostic factors for NSCLC patients.

Use of serum circulating CCNB2 in cancer surveillance.

Cyclin B2 (CCNB2), a member of the cyclin protein family, has been found to be up-regulated in human cancers. To evaluate the potential use of circulating CCNB2 in serum in cancer surveillance, we examined relative expression levels of serum circulating CCNB2 mRNA in 103 cancer patients, 19 normal controls, and 40 benign disease patients using real-time quantitative reverse transcriptase polymerase chain reaction. We found that the relative expression level of circulating CCNB2 mRNA in cancer patients was significantly higher (p<0.0001) than that in normal controls and benign diseases group. Circulating CCNB2 mRNA level was significantly (p<0.001) correlated with cancer stage and metastasis status. Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.87 and 0.83 (p<0.05) in identifying cancer patients' metastasis status in lung and digestive tract cancer, respectively. Moreover, we observed that expression levels of circulating CCNB2 mRNA in cancer patients significantly decreased (p=0.0084) after their therapeutic treatments. These data suggest that detection of serum circulating CCNB2 mRNA may have potential clinical applications in screening and monitoring of metastasis and therapeutic treatments.

Use of serum circulating CCNB2 in cancer surveillance.

Cyclin B2 (CCNB2), a member of the cyclin protein family, has been found to be up-regulated in human cancers. To evaluate the potential use of circulating CCNB2 in serum in cancer surveillance, we examined relative expression levels of serum circulating CCNB2 mRNA in 103 cancer patients, 19 normal controls, and 40 benign disease patients using real-time quantitative reverse transcriptase polymerase chain reaction. We found that the relative expression level of circulating CCNB2 mRNA in cancer patients was significantly higher (p<0.0001) than that in normal controls and benign diseases group. Circulating CCNB2 mRNA level was significantly (p<0.001) correlated with cancer stage and metastasis status. Receiver operating characteristic (ROC) analysis showed an area under the curve (AUC) of 0.87 and 0.83 (p<0.05) in identifying cancer patients' metastasis status in lung and digestive tract cancer, respectively. Moreover, we observed that expression levels of circulating CCNB2 mRNA in cancer patients significantly decreased (p=0.0084) after their therapeutic treatments. These data suggest that detection of serum circulating CCNB2 mRNA may have potential clinical applications in screening and monitoring of metastasis and therapeutic treatments.