Although aberrant microRNA (miRNA) is expressed in different types of human cancer tissues, its pathophysiologic role and the relevance of tumorigenesis and metastasis are still largely unknown. ...Here, we defined miRNAs involved in cancer metastasis (metastamirs) using an established mouse model for peritoneal dissemination of human scirrhous gastric carcinoma cells. Highly metastatic derivatives (44As3 cells) were derived from the parental cells originally isolated from patients (HSC-44PE cells). Using microarray analysis to identify differentially expressed miRNAs in 44As3 and HSC-44PE cells, we focused on miR-516a-3p as a candidate antimetastatic miRNA (antimetastamir) whose functions in cancer had not been studied. We confirmed attenuated expression of miR-516a-3p in 44As3 cells compared with HSC-44PE cells by Northern blot analysis and quantitative reverse transcriptase PCR. Stable ectopic overexpression in 44As3-miR-516a-3p cells permitted identification of sulfatase 1 as a direct target of the miRNA, through use of the isobaric tagging reagent iTRAQ and the QSTAR Elite Hybrid LC-MS/MS system. Sulfatase 1 is known to remove 6-O-sulfates from heparan sulfate proteoglycans on the cell surface, causing release of membrane-bound Wnt ligands from cells. Consistent with this function, Western blot analyses revealed high levels of Wnt3a, Wnt5a, and nuclear β-catenin accumulation in 44As3 cells but relatively reduced levels in 44As3-miR-516a-3p cells. Notably, orthotopic inoculation of nude mice with 44As3-miR-516a-3p cells yielded significantly longer survival periods compared with mice inoculated with control 44As3 cells. Through atelocollagen-mediated delivery of an miR-516a-3p expression vector into orthotopic 44As3 tumors, we documented its feasibility as a treatment agent. Our findings define the miRNA miR-516-3p as an antimetastamir with potential therapeutic applications in blocking metastatic dissemination of gastric cancers.
Aggressive periodontitis and NOD2 variants Mizuno, Noriyoshi; Kume, Kodai; Nagatani, Yukiko ...
Journal of human genetics,
10/2020, Letnik:
65, Številka:
10
Journal Article
Recenzirano
Aggressive periodontitis (AgP) occurs at an early age and causes rapid periodontal tissue destruction. Nucleotide-binding oligomerization domain-containing protein 2 (NOD2) encodes a protein with two ...caspase recruitment domains and eleven leucine-rich repeats. This protein is expressed mainly in peripheral blood leukocytes and is involved in immune response. NOD2 variants have been associated with increased susceptibility to Crohn's disease, and recently, NOD2 was reported as a causative gene in AgP. The present study aimed to identify potential NOD2 variants in an AgP cohort (a total of 101 patiens: 37 patients with positive family histories and 64 sporadic patients). In the familial group, six patients from two families had a reported heterozygous missense variant (c.C931T, p.R311W). Four patients in the sporadic group had a heterozygous missense variant (c.C1411T, p.R471C), with no reported association to the disease. Overall, two NOD2 variants, were identified in 10% of our AgP cohort. These variants were different from the major variants reported in Crohn's disease. More cases need to be investigated to elucidate the role of NOD2 variants in AgP pathology.
Methylation, the most common chemical modification of cellular components such as DNA, RNA, and proteins, impacts biological processes including transcription, RNA processing, and protein dynamics. ...Although abnormal expression of methyltransferase can lead to various diseases including cancers, little is known about the relationship between methyltransferase and cancers. Here we aimed to understand the role of methyltransferase in cancer metastasis. We found that elevated methyltransferase-like 9 (METTL9) is closely associated with the acquisition of metastatic activity in human scirrhous gastric cancers. The stable knockdown of METTL9 via an shRNA vector technique in our original metastatic cells from scirrhous gastric cancer patients significantly inhibited migration and invasion. In metastatic cells, METTL9 protein is predominantly localized in mitochondria, and the METTL9 knockdown significantly reduced mitochondrial Complex I activity. METTL9 can be a candidate of molecular targets to inhibit peritoneal dissemination of scirrhous gastric cancers. This report is the first to describe the relationship between METTL9 and cancer metastasis.
•Elevated METTL9 correlates with metastasis in human scirrhous gastric cancer.•This is the first report on the biological relationship between METTL9 and metastasis.•METTL9 protein localizes mainly in mitochondria in metastatic scirrhous gastric cancer.•METTL9 knockdown reduces mitochondrial Complex I activity to decrease cell migration and invasion in metastatic scirrhous gastric cancer.•METTL9 holds promise against peritoneal dissemination.
In this study, we synthesized Dicer‐substrate siRNA conjugated with palmitic acid at the 5′‐end of the sense strand (C16‐DsiRNA), and examined its RNAi effect on β‐catenin as a target gene in a colon ...cancer cell line, HT29Luc, both in vitro and in vivo. We examined the in vitro RNAi effect in HT29Luc cells and found that C16‐DsiRNA strongly inhibited expression of the β‐catenin gene in comparison with non‐modified DsiRNA. Also, high membrane permeability of C16‐DsiRNA was exhibited, and it was confirmed that most of the C16‐DsiRNA was localized in cytoplasm of HT29Luc cells. In regard to the in vivo RNAi effect, C16‐DsiRNA complexed with Invivofectamine targeting the β‐catenin gene was locally administered to a subcutaneous tumor formed by implantation of HT29Luc cells into the subcutis of nude mice; we evaluated the effect by measuring the bioluminescence increase, which reflects tumor growth, using an in vivo imaging system. As a result, C16‐DsiRNA strongly inhibited the growth of tumors formed in subcutis of nude mice compared with non‐modified DsiRNA, and this in vivo RNAi effect lasted up to 15 days. Our results suggest that C16‐DsiRNA should be vigorously pursued as a novel nucleic acid medicine for clinical treatment of cancer.
We investigated the in vitro and in vivo RNAi efficacy in colon cancer cells of C16‐DsiRNA targeting endogenous β‐catenin, which is active during cancer cell development and is associated with the growth of cancer. C16‐DsiRNA exhibited a strong target gene silencing effect, high cellular uptake, and excellent nuclease resistance in vitro. Likewise, C16‐DsiRNA showed strong inhibition of the growth of tumors formed in our subcutaneous tumor mouse model, and this in vivo RNAi effect lasted up to 15 days.
Patients with B cell lymphomas bearing MYC translocation combined with translocation involving other genes, such as BCL2, BCL3, or BCL6, defined as double-hit lymphoma (DHL), have a poor prognosis. ...Recent studies expanded the concept to include double-expressing lymphoma (DEL) that co-overexpresses MYC protein with either of those proteins. Accordingly, we defined cytogenetic DHL and DEL as primary DHL. An adoptive T cell immunotherapy with a chimeric antigen receptor (CAR) has been clinically shown to exhibit cytotoxicity in refractory neoplasias. We revealed the marked cytotoxicity of anti-CD19- and/or anti-CD38-CAR T cells against primary DHL cells from patients. CD19- and/or CD38-specific T cells were co-cultured with cytogenetic DHL (n = 3) or DEL (n = 2) cells from five patients for 3 days. We examined whether T cells retrovirally transduced with each vector showed cytotoxicity against DHL cells. Anti-CD19- and/or anti-CD38-CAR T cells were co-cultured with primary DHL cells at an E:T ratio of 1:2 for 3 days. Anti-CD19- and anti-CD38-CAR T cells completely abrogated these DHL cells, respectively. Anti-CD19-CAR T cells synergistically exerted collaborative cytotoxicity against these primary DHL cells with anti-CD38-CAR T cells. Therefore, refractory DHL cells can be efficiently abrogated by the clinical use of T cells with anti-CD19- and/or anti-CD38-CAR.
We herein report the case of a 60-year-old man with a "target sign" in the left frontal lobe on magnetic resonance imaging (MRI), which is thought to be a specific sign of cerebral toxoplasmosis. ...18F-fluorodeoxyglucose-positron emission tomography showed no increased uptake, and 201Tl-single photon emission computed tomography showed the focal uptake in the left frontal lesion. On a brain biopsy, the patient was given a definitive diagnosis of brain metastasis from diffuse large B-cell lymphoma, and cerebral toxoplasmosis was excluded. In the present case, multilayer intensities on MRI may reflect the fast-growing nature of this tumor.
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