Molecular mechanism of MLCK1 inducing 5-Fu resistance in colorectal cancer cells through activation of TNFR2/NF-κB pathway.

BACKGROUND AND AIMS: Chemotherapy resistance in colorectal cancer have been faced with significant challenges in recent years. Particular interest is directed to tumor microenvironment function. Recent work has, identified a small molecule named Divertin that prevents myosin light chain kinase 1(MLCK1) recruitment to the perijunctional actomyosin ring(PAMR), restores barrier function after tumor necrosis factor(TNF)-induced barrier loss and prevents disease progression in experimental inflammatory bowel disease. Studies have shown that MLCK is a potential target for affecting intestinal barrier function, as well as for tumor therapy. However, the relative contributions of MLCK expression and chemotherapy resistance in colorectal cancers have not been defined. METHODS: Statistical analysis of MYLK gene expression differences in colorectal cancer patients and normal population and prognosis results from The Cancer Genome Atlas(TCGA) data. Cell activity was detected by Cell counting Kit-8. Cell proliferation was detected by monoclonal plate. The apoptosis was detected by flow cytometry and western blot. Determine the role of MLCK1 in inducing 5-Fluorouracil(5-Fu) resistance in colorectal cancer cells was detected by overexpression of MLCK1 and knock-down expression of MLCK1. RESULTS: MLCK1 is expressed at different levels in different colorectal cancer cells, high MLCK1 expressing cell lines are less sensitive to 5-Fu, and low MLCK1 expressing cell lines are more sensitive to 5-Fu. MLCK1 high expression enhances resistance to 5-Fu in colorectal cancer cells and the sensitivity to 5-Fu was increased after knocking down the expression of MLCK1, that might be closely correlated to TNFR2/NF-κB pathway. CONCLUSIONS: MLCK1 high expression can enhance resistance to 5-Fu in colorectal cancer cells and the sensitivity to 5-Fu was increased after knocking down the expression of MLCK1, that might be closely correlated to TNFR2/NF-κB pathway, which will provide a new method for the treatment of colorectal cancer patients who are resistant to 5-Fu chemotherapy.

Interferon Inducing Porcine Reproductive and Respiratory Syndrome Virus Vaccine Candidate Protected Piglets from HP-PRRSV Challenge and Evoke a Higher Level of Neutralizing Antibodies Response.

Although widespread administration of attenuated porcine reproductive and respiratory syndrome virus (PRRSV) vaccines has been implemented since they first became commercially available two decades ago, PRRSV infection prevalence in swine herds remains high. The limited success of PRRSV vaccines is partly due to the well-established fact that a given vaccine strain confers only partial or no protection against heterologous strains. In our past work, A2MC2-P90, a novel PRRSV vaccine candidate that induced a type I IFNs response in vitro, conferred complete protection against challenge with genetically heterologous PRRSV strains. Here we assessed the ability of the PRRSV vaccine candidate A2MC2-P90 to protect piglets against the HP-PRRSV challenge and compared its efficacy to that of a licensed HP-PRRSV-specific vaccine (TJM-F92) assessed in parallel. A2MC2-P90 provided vaccinated piglets with 100% protection from a lethal challenge with extremely virulent HP-PRRSV-XJA1, while 100% mortality was observed for unvaccinated piglets by day 21 post-challenge. Notably, comparison of partial sequence (GP5) of XJA1 to A2MC2-P90 suggested there was only 88.7% homology. When comparing post-HP-PRRSV challenge responses between piglets administered A2AMC2-P90 versus those immunized with licensed vaccine TJM-F92, A2MC2-P90-vaccinated piglets rapidly developed a stronger protective humoral immune response, as evidenced by much higher titers of neutralizing antibodies, more rapid clearance of viremia and less nasal virus shedding. In conclusion, our data suggest that this novel vaccine candidate A2MC2-P90 has improved protection spectrum against heterologous HP-PRRSV strains.

Echinocandins versus Amphotericin B Against Candida tropicalis Fungemia in Adult Hematological Patients with Neutropenia: A Multicenter Retrospective Cohort Study.

BACKGROUND: Candida tropicalis is the most common non-albicans Candida species identified in immunocompromised patients, which often appears with high mortality. However, data on the outcomes of treatment for Candida tropicalis fungemia in patients with neutropenia remain limited. METHODS: In the present study, 90 neutropenic adult patients with proven Candida tropicalis fungemia, who received initial antifungal therapy, were retrospectively analyzed. RESULTS: These results revealed that the overall 8-day and 30-day mortality among patients in the entire data set were 22.2% and 33.3%, respectively. However, there was no significant difference between the survival and death group, in terms of baseline characteristics. The univariate analysis of risk factors identified the treatment with azole as a predictor of mortality, while treatments that containing amphotericin B were associated with reduced mortality. In addition, the survival rate on day 30 was observed in 60.7% (17/28) of patients who were initially treated with echinocandins, while this was observed in 86.4% (19/22, P=0.039) and 100% (13/13, P=0.024) of patients treated with amphotericin B plus echinocandins and amphotericin B, respectively. CONCLUSION: These data indicate for the first time that the initial therapy with amphotericin B-based agents was associated with a better survival rate and could be assessed as the optimal strategy for the treatment of Candida tropicalis fungemia in patients with neutropenia.

Erianin, a novel dibenzyl compound in Dendrobium extract, inhibits lung cancer cell growth and migration via calcium/calmodulin-dependent ferroptosis.

Ferroptosis, a novel form of programmed cell death, is characterized by iron-dependent lipid peroxidation and has been shown to be involved in multiple diseases, including cancer. Stimulating ferroptosis in cancer cells may be a potential strategy for cancer therapy. Therefore, ferroptosis-inducing drugs are attracting more attention for cancer treatment. Here, we showed that erianin, a natural product isolated from Dendrobium chrysotoxum Lindl, exerted its anticancer activity by inducing cell death and inhibiting cell migration in lung cancer cells. Subsequently, we demonstrated for the first time that erianin induced ferroptotic cell death in lung cancer cells, which was accompanied by ROS accumulation, lipid peroxidation, and GSH depletion. The ferroptosis inhibitors Fer-1 and Lip-1 but not Z-VAD-FMK, CQ, or necrostatin-1 rescued erianin-induced cell death, indicating that ferroptosis contributed to erianin-induced cell death. Furthermore, we demonstrated that Ca2+/CaM signaling was a critical mediator of erianin-induced ferroptosis and that blockade of this signaling significantly rescued cell death induced by erianin treatment by suppressing ferroptosis. Taken together, our data suggest that the natural product erianin exerts its anticancer effects by inducing Ca2+/CaM-dependent ferroptosis and inhibiting cell migration, and erianin will hopefully serve as a prospective compound for lung cancer treatment.

Clinicopathologic significance and therapeutic implication of de novo CD5+ diffuse large B-cell lymphoma.

BACKGROUND: Various subsets of diffuse large B-cell lymphoma(DLBCL) are distinguished based on molecular and immunohistochemical features. As we know, CD5 is a pan-T-cell surface marker and is seldom expressed in DLBCL. Large-scale studies of de novo CD5+ DLBCL are lacking in Chinese patients. METHOD: A total of 139 patients with DLBCL (30 CD5+ DLBCL and 109 CD5- DLBCL) who were immunophenotyped and treated with chemotherapy were subjected to this analysis. There were 85 males and 54 females. Their age ranged from 17 to 84 years old, and the median age was 58 years old. RESULTS: In this study CD5+ DLBCL was associated with higher IPI scores (>2), bone marrow involvement, higher probability of >1 ECOG performance status, non-germinal center B-cell like(non-GCB), BCL2 overexpression, whereas seldom expressed CD10 or BCL6, and unconspicuous higher expression of Ki67. With standard chemotherapy, CD5+ DLBCL patients had significantly worse overall survival (OS, median, 29.5 months vs. not reached, P = 0.0004) and progression-free survival (PFS, median, 18.0 months vs. not reached, P = 0.0002) than CD5- DLBCL patients, which had independent prognostic significance of the International Prognostic Index (IPI), and subtype of the non-GCB DLBCL. For CD5+ DLBCL, the addition of rituximab to chemotherapy may not significantly improve the OS (median, 14 months vs. 29.5 months, P = 0.72) and PFS (median, 10 months vs. 12 months, P = 0.92). CONCLUSION: CD5+ DLBCL patients have the distinctive clinical and biological features, they should be provided with clinic individualized treatment and important pathways with therapeutic implications should be underscored.

Electrochemical Visualization of Intracellular Hydrogen Peroxide at Single Cells.

In this Letter, the electrochemical visualization of hydrogen peroxide inside one cell was achieved first using a comprehensive Au-luminol-microelectrode and electrochemiluminescence. The capillary with a tip opening of 1-2 µm was filled with the mixture of chitosan and luminol, which was coated with the thin layers of polyvinyl chloride/nitrophenyloctyl ether (PVC/NPOE) and gold as the microelectrode. Upon contact with the aqueous hydrogen peroxide, hydrogen peroxide and luminol in contact with the gold layer were oxidized under the positive potential resulting in luminescence for the imaging. Due to the small diameter of the electrode, the microelectrode tip was inserted into one cell and the bright luminescence observed at the tip confirmed the visualization of intracellular hydrogen peroxide. The further coupling of oxidase on the electrode surface could open the field in the electrochemical imaging of intracellular biomolecules at single cells, which benefited the single cell electrochemical detection.