Prognostic evaluation of a multi-target magnetic bead-enriched circulating tumor cell-enriched identification system for colorectal cancer.

Background: Colorectal cancer (CRC) is the third most prevalent cancer in the world. Traditional tissue biopsy cannot provide dynamic monitoring of patients' tumors or reflect the characteristics of tumors in real time because the sampling process is invasive and accompanied by risks. Circulating tumor cells (CTCs) are considered a major cause of tumor metastasis, and investigating CTCs helps to understand the biology and vulnerability of malignant tumors during hematogenous metastasis. Methods: We sequentially used epithelial cell adhesion molecule (EpCAM)-coated immunoliposomal magnetic beads (Ep-IMBs) and vimentin-coated immunoliposomal magnetic beads (Vi-IMBs) to capture and characterize CTCs from 110 CRC patients. We further constructed a Cox risk regression model, optimized the model composition using backward stepwise regression, and finally applied nomograms to show the effect of each variable on survival risk. Results: The specificity of the CTCs enrichment and identification system was 100% and the sensitivity was 79.0%. Multivariate analysis indicated total CTC number was an important predictor for bad survival, whereas American Joint Committee on Cancer (AJCC) stage, lymph node metastasis, and carcinoembryonic antigen (CEA) level were associated with prognosis, and the risk of mortality was associated with the AJCC stage of the CRC. Conclusions: The CTC enrichment and identification system constructed in this research demonstrated superior accuracy. In addition, CTCs can be used as an important predictor for prognosis of patients with CRC, and the combination of other clinical predictive factors can help clinicians to better design individualized treatment regimens, which is of great clinical application value.

Dynamic detection of HER2 of circulating tumor cells in patients with gastric carcinoma and its clinical application.

The aim of the present study was to construct and characterize human epidermal growth factor receptor 2 (HER2) lipid magnetic ball (H­LMB) for separating circulating tumor cells (CTCs) in patients with gastric carcinoma (GC) and to compare the result of separated CTC counts with that of next­generation sequencing (NGS) for single­gene analysis to verify the consistency for evaluating the association between the detection results and the progress of clinical treatment, so as to facilitate early diagnosis and dynamic monitoring of GC. A lipid magnetic ball (LMB), coated with Fe3O4 nanoparticles, was synthesized by microemulsion technique and an anti­HER2 antibody was conjugated to the surface of LMB to form H­LMB, followed by the characterization of the prepared H­LMB. The detection of capture efficiency of LMBs in GC cells was tested by MTT and expression of HER2 mRNA was determined by reverse transcription­quantitative PCR. The positive detection rate of HER2 was verified by HER2­fluorescence in situ hybridization (FISH) test on the separated CTCs from GC. Further verification was performed based on the consistency between the result of separated CTCs and that of single­gene NGS assay of HER2, associated with the determination of clinical consistency. The constructed H­LMB exhibited good stability and specificity. The mutation rate of HER2 by the FISH test was 14% in the blood samples of 50 patients with GC and was 14% by NGS assay. The mutation rate of HER2 was 12% in H­LMB and the positive detection rate was 85.7% compared with the results of the FISH test, indicating consistency with the clinical diagnosis and pathological examination results. In conclusion, the anti­HER2 antibody­modified LMB can separate CTCs with HER2 abnormal expression, which exhibits an application potential in GC diagnosis and treatment and is of great clinical significance for the diagnosis and evaluation of its therapeutic effect on GC.

Distribution and Clinical Analysis of EpCAM+/Vimentin+ Circulating Tumor Cells in High-Risk Population and Cancer Patients.

Circulating Tumor Cells (CTCs) are already present in the peripheral blood of patients with early tumors and even precancerous lesions. The objective of this study was to determine the count of CTCs in peripheral blood from high-risk population(HRP), healthy subjects and patients with Pan-cancer. The CTCs in the peripheral blood from HRP and cancer patients were enriched and identified based on the positive sorting method by epithelial cell adhesion molecular (EpCAM) liposome magnetic bead (Ep-LMB) and Vimentin liposome magnetic bead (Vi-LMB). Simultaneously, further analysis was carried out focusing on the clinical characteristics of patients by collecting the peripheral blood samples from healthy subjects as the parallel control. According to the results, the prepared LMBs had high specificity and stability, resulting in an average (Av) proliferation rate of over 90% for each cell line, and the average capture rate of higher than 80%. In terms of CTCs count detection in clinical blood samples, the average count was 0.9 (Ep: Av=0.6, Vi: Av=0.3), 2.4 (Ep: Av=1.4, Vi: Av=0.8) and 7.3 (Ep: Av=4.0, Vi: Av=3.3) in healthy subjects, HRP and total cancer patients, respectively. Besides, there was no obvious difference in the average count of CTCs among patients with different cancer types. While count of CTCs in the aforementioned cancer patients was statistically different from that in healthy subjects and patients with HRP. The survival time of cancer patients whose number of CTCs is greater than the average is significantly increased. Collectively, the study confirmed that CTCs can achieve early tumor detection and auxiliary diagnosis, and its number is related to the occurrence and development of tumors, and CTCs can be detected in HRP and sub-health population.

MiR-130b inhibits proliferation and induces apoptosis of gastric cancer cells via CYLD.

A role of microRNA-130b (miR-130b) in the carcinogenesis of gastric cancer remains undetermined. In this study, we studied the effects and mechanism of miR-130b to the gastric cell proliferation and apoptosis. We found that the levels of miR-130b significantly up-regulated in gastric cancer tissue, compared to the paired adjacent non-tumor gastric tissue. The miR-130b levels in gastric cancer cell lines were significantly higher than those in control normal gastric tissues. Transfection with the miR-130b mimic enhanced the cell proliferation and suppressed cell apoptosis in gastric cancer cells, while transfection with the anti-sense of miR-130b (anti-miR-130b) suppressed cell proliferation and induced cell apoptosis in gastric cancer cells. Bioinformatics analyses showed that cylindromatosis gene (CYLD) was a potential target gene of miR-130b. The luciferase activity assay and western blot verified that miR-130b targeted CYLD messenger RNA (mRNA) to modulate its protein levels. Together, our study suggests that aberrantly expressed miR-130b may regulate cell apoptosis and proliferation of human gastric cancer cells via CYLD, which appears to be a promising therapeutic target for gastric cancer.

SUMO-specific protease 6 promotes gastric cancer cell growth via deSUMOylation of FoxM1.

SUMOylation is a post-translational modification exerted various effects on the target proteins. SUMOylation is a highly dynamic and reversible process, which has been shown to play an important role in tumorigenesis. However, the roles of sentrin/SUMO-specific proteases (SENPs), which mediate the reverse process of SUMOylation, in tumorigenesis remains largely unexplored. Here, we uncover a critical role of SENP6 in promoting gastric cancer cells growth via regulating the deSUMOylation of a transcription factor forkhead box protein M1 (FoxM1). We demonstrated that the mRNA and protein levels were elevated in gastric cancer tissues. Overexpression of SENP6 promoted, while RNA interference depletion of endogenous SENP6 inhibited gastric cancer cells growth and the ability of colony formation. By using biochemical assays, we identified FoxM1 as a novel substrate of SENP6 in gastric cancer cells. Thus, our data suggest that SENP6, which is highly expressed in gastric cancer cells, regulates the transcriptional activity and stability of FoxM1 through deSUMOylation.

Expression of group IIA phospholipase A2 is an independent predictor of favorable outcome for patients with gastric cancer.

Growing evidence suggests that phospholipase A2 (PLA2) plays a pivotal role in tumorigenesis in human gastrointestinal cancer. One of the well-studied isoforms of PLA2, group IIA PLA2 (PLA2G2A), appears to exert its protumorigenic or antitumorigenic effects in a tissue-specific manner. The present study was designed to determine the expression profile and prognostic value of PLA2G2A in gastric cancer in a large Chinese cohort. By using real-time polymerase chain reaction, the amount of PLA2G2A messenger RNA in 60 pairs of fresh gastric tumors and adjacent noncancerous mucosa was measured. The immunostaining of PLA2G2A in 866 gastric cancers with paired noncancerous tissues was assayed. No expression of PLA2G2A was found in normal gastric mucosa, and focal expression of PLA2G2A was noticed in intestinal metaplasia, whereas significantly increased expression of PLA2G2A was observed in the cytoplasm of gastric cancer cells. Furthermore, the extent of PLA2G2A expression was associated with tumor size (P < .001), tumor differentiation (P = .001), T class (P < .001), N class (P < .001), and TNM stage (P < .001) of gastric cancer. Multivariate analysis showed that PLA2G2A expression was an independent predictor of survival for patients with gastric cancer (P = .024). Expression of PLA2G2A seems to be protective for patients with gastric cancer (hazard ratio, 1.423; 95% confidence interval, 1.047-1.935), and it may be a target for achieving better treatment outcomes.

[Clinical values of detecting excision repair cross complementing 1 and top-isomerase I in individualized therapies of metastatic colorectal cancer].

OBJECTIVE: To explore the clinical values of detecting drug related molecules excision repair cross complementing 1 (ERCC1) and top-isomerase I (TOPO I) in individualized therapies of metastatic colorectal cancer. METHODS: From June 2009 to December 2011, 90 patients at Huadong Hospital with metastatic colorectal cancer were randomly separated into 2 groups after operation. Each group had 45 patients without difference in gender, age or TNM stage. The expressions of ERCC1 and TOPO Iin cancer tissues were detected by immunohistochemical staining. The testing group received individualized chemotherapies following the expression results while the control group had random chemotherapies. The survival difference between two groups was analyzed by log-rank test and Kaplan-Meier analysis. And curative effect was analyzed by χ(2) or Fisher's analysis. RESULTS: The expressions of ERCC1 and TOPO I had no statistical significance between two groups (both P > 0.05). In the testing group, the median survival time was 281 days and the beneficial ratio 51.1% (23/45) versus 246 days and 44.4% (20/45) respectively in the control group. The inter-group comparisons of survival (P = 0.235) and curative effect (χ(2) = 0.04, P > 0.05) showed no statistical significance. In the estimated drug tolerated group (ERCC1 high expression or TOPO I low expression), the median survival time was 196 days and the beneficial ratio 4/14 versus 304 days and 51.3% (39/76) in the estimated drug sensitive group. The inter-group comparisons of survival and curative effect (both P < 0.05) had statistical significance. The median survival time and beneficial ratio significantly increased in estimated drug sensitive group than those in estimated drug tolerated group. CONCLUSIONS: The expression of drug related molecule in colorectal cancer tissue is significantly associated with curative effect in patients. Patients with down-regulated ERCC1 on Oxaliplatin or up-regulated TOPO Ion Irinotecan have longer survival and better curative effect. And chemotherapies guided by drug related molecule detection may boost curative effects in metastatic colorectal cancer.

Therapeutic efficacy of high-dose vitamin C on acute pancreatitis and its potential mechanisms.

AIM: To observe the therapeutic efficacy of high-dose Vitamin C (Vit. C) on acute pancreatitis (AP), and to explore its potential mechanisms. METHODS: Eighty-four AP patients were divided into treatment group and control group, 40 healthy subjects were taken as a normal group. In the treatment group, Vit. C (10 g/day) was given intravenously for 5 days, whereas in the control group, Vit. C (1 g/day) was given intravenously for 5 days. Symptoms, physical signs, duration of hospitalization, complications and mortality rate were monitored. Meanwhile, serum amylase, urine amylase and leukocyte counts were also determined. The concentration of plasma vitamin C (P-VC), plasma lipid peroxide (P-LPO), plasma vitamin E (P-VE), plasma beta-carotene (P-beta-CAR), whole blood glutathione (WB-GSH) and the activity of erythrocyte surperoxide dimutase (E-SOD) and erythrocyte catalase (E-CAT) as well as T lymphocyte phenotype were measured by spectrophotometry in the normal group and before and after treatment with Vit. C in the treatment and the control group. RESULTS: Compared with the normal group, the average values of P-VC, P-VE, P-beta-CAR, WB-GSH and the activity of E-SOD and E-CAT in AP patients were significantly decreased and the average value of P-LPO was significantly increased, especially in severe acute pancreatitis (SAP) patients (P<0.05. P-VC, P=0.045; P-VE, P=0.038; P=0.041; P-beta-CAR, P=0.046; WB-GSH, P=0.039; E-SOD, P=0.019; E-CAT, P=0.020; P-LPO, P=0.038). Compared with the normal group, CD3 and CD4 positive cells in AP patients were significantly decreased. The ratio of CD4/CD8 and CD4 positive cells were decreased, especially in SAP patients (P<0.05. CD4/CD8, P=0.041; CD4, P =0.019). Fever and vomiting disappeared, and leukocyte counts and amylase in urine and blood become normal quicker in the treatment group than in the control group. Moreover, patients in treatment group also had a higher cure rate, a lower complication rate and a shorter in-ward days compared with those in he control group. After treatment, the average value of P-VC was significantly higher and the values of SIL-2R, TNF-alpha, IL-6 and IL-8 were significantly lower in the treatment group than in the control group (P<0.05 P-VC, P=0.045; SIL-2R, P=0.012; TNF-alpha, P=0.030; IL-6, P=0.015; and IL-8, P=0.043). In addition, the ratio of CD4/CD8 and CD4 positive cells in the patients of treatment group were significantly higher than that of the control group after treatment (P<0.05. CD4/CD8, P=0.039; CD4, P=0.024). CONCLUSION: High-dose vitamin C has therapeutic efficacy on acute pancreatitis. The potential mechanisms include promotion of anti-oxidizing ability of AP patients, blocking of lipid peroxidation in the plasma and improvement of cellular immune function.