ABSTRACT
Objective:To investigate the role of NOD-like receptor protein 3 (NLRP3) in the regulation of phagocytosis in Vibrio vulnificus ( V. vulnificus)-infected macrophages. Methods:Expression profiles of phagocytosis-related genes in PBS- and V. vulnificus-infected J774A.1 cells were analyzed by RNA-Seq. NLRP3-knockout (NLRP3 KO) J774A.1 cells were constructed using CRISPR-Cas9 gene-editing system. The phagocytosis of V. vulnificus and pHrodo RED-labelled Escherichia coli ( E. coli) bioparticles in parental and NLRP3 KO J774A.1 cells was detected by flow cytometry. Real-time PCR was performed to measure the expression of Fgr2 b gene at mRNA level in PBS- and V. vulnificus-treated parental and NLRP3 KO J774A.1 cells. Results:The expression of 18 phagocytosis-related genes was upregulated in V. vulnificus-infected J774A.1 cells than in PBS-treated J774A.1 cells ( P<0.05). There was a 5 bp deletion in the exon 2 of NLRP3 gene in NLRP3 KO J774A.1 cells, resulting in frameshift mutation and complete loss of NLRP3 expression. NLRP3 KO J774A.1 cells exhibited enhanced phagocytosis of V. vulnificus and pHrodo RED-labelled E. coli bioparticles than parental J774A.1 cells ( P<0.05). Besides, the expression of Fgr2 b gene at mRNA level was significantly increased in V. vulnificus-infected NLRP3 KO J774A.1 cells than in parental J774A.1 cells ( P<0.05). Conclusions:The phagocytosis of V. vulnificus in macrophages could be negatively regulated by NLRP3, which was possibly mediated through the regulation of Fgr2 b gene expression.
ABSTRACT
The present study aimed to investigate the efficacy of five signaling pathway inhibitors, N-[N-(3,5difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester, vismodegib, salinomycin, ruxolitinib and stattic, as novel therapeutic agents that target breast cancer stem cells (BCSCs) in triple-negative breast cancer (TNBC). The in vitro anti-proliferative, anti-invasive, pro-apoptotic and inhibitory effects on BCSC self-renewal of these signaling pathway inhibitors on the TNBC stem cell line HCC38 were examined by MTT assays, Matrigel invasion assays, flow cytometry and suspension mammosphere assays, respectively. For the in vivo study, another TNBC stem cell line, HCC1806, pretreated with these signaling pathway inhibitors, was inoculated into female nonobese diabetic/severe combined immunodeficient mice, and the tumor volumes were measured following tumor formation. Treatment of HCC38 cells with each signaling pathway inhibitor significantly decreased TNBC cell proliferation, cell invasion and mammosphere formation while inducing cell apoptosis by inhibiting the protein expression or phosphorylation of downstream signaling molecules. In the xenograft mouse models, tumor formation and growth of HCC1806 cells pretreated with each signaling pathway inhibitor were effectively suppressed. Treatment with these signaling pathway inhibitors may provide a novel therapeutic strategy against TNBC by targeting BCSCs, thus providing promising insight for clinical applications in patients with TNBC.
Subject(s)
Antineoplastic Agents/pharmacology , Neoplastic Stem Cells/drug effects , Signal Transduction/drug effects , Triple Negative Breast Neoplasms/economics , Animals , Apoptosis/drug effects , Cell Line, Tumor , Cell Movement/drug effects , Cell Proliferation/drug effects , Disease Models, Animal , Female , Humans , Mice , Mice, Inbred NOD , Mice, SCID , Tumor Burden/drug effects , Xenograft Model Antitumor Assays/methodsABSTRACT
OBJECTIVE: We evaluated the effects of epidermal growth factor (EGF) and ofloxacin otic drops on the healing of large human traumatic tympanic membrane perforations (TMPs). STUDY DESIGN: Case series with chart review. SETTING: Tertiary university hospital. METHODS: Retrospective case review of patients with traumatic TMP larger than 25% of the TM seen between February 2007 and December 2008. Patients were stratified into EGF drops, ofloxacin drops, and observation groups. The closure rate, closure time, and hearing level were compared among the three groups at 6months. RESULTS: In total, 120 patients met the inclusion criteria. The total closure rate was 89.2% (107/120) and the total mean closure time was 22.6±7.4days. The closure rates of perforation in the EGF, ofloxacin otic drops, and observation groups were 93.5%, 92.0%, and 82.2%, respectively. The closure rates among the three groups were not statistically different (p=0.19). The mean perforation closure times were 12.6±6.9, 12.9±5.1, and 35.7±9.2days for the EGF, ofloxacin otic drops, and observation groups, respectively. The average closure time in the observation group was significantly longer (p=0.01) than that in the EGF and ofloxacin otic drops groups. However, the closure times in the EGF and ofloxacin otic drops groups were not significantly different (p=0.84). CONCLUSIONS: The study surprisingly found that both topical application of EGF and ofloxacin otic drops result in more rapid closure compared with spontaneous healing for human large traumatic TMPs. The benefit would be great as a shorter recovery time may reduce health care costs. Therefore, ofloxacin otic drops should be considered in clinics.
