S100P Expression via DNA Hypomethylation Promotes Cell Growth in the Sessile Serrated Adenoma/Polyp-Cancer Sequence

Digestion ◽  
2021 ◽  
pp. 1-14
Author(s):  
Sayo Takahashi ◽  
Koichi Okamoto ◽  
Toshihito Tanahashi ◽  
Shota Fujimoto ◽  
Tadahiko Nakagawa ◽  
...  
Keyword(s):  
Cell Growth ◽  
Dna Hypomethylation ◽  
Rt Pcr ◽  
Serrated Adenoma ◽  
Methylation Array ◽  
Expression Levels ◽  
3 Dimensional ◽  
Cpg Sites ◽  
Shrna Vector ◽  
The Mean

<b><i>Background/Aims:</i></b> Sessile serrated adenomas/polyps (SSA/Ps) are a putative precursor lesion of colon cancer. Although the relevance of DNA hypermethylation in the SSA/P-cancer sequence is well documented, the role of DNA hypomethylation is unknown. We investigated the biological relevance of DNA hypomethylation in the SSA/P-cancer sequence by using 3-dimensional organoids of SSA/P. <b><i>Methods:</i></b> We first analyzed hypomethylated genes using datasets from our previous DNA methylation array analysis on 7 SSA/P and 2 cancer in SSA/P specimens. Expression levels of hypomethylated genes in SSA/P specimens were determined by RT-PCR and immunohistochemistry. We established 3-dimensional SSA/P organoids and performed knockdown experiments using a lentiviral shRNA vector. DNA hypomethylation at CpG sites of the gene was quantitated by MassARRAY analysis. <b><i>Results:</i></b> The mean number of hypomethylated genes in SSA/P and cancer in SSA/P was 41.6 ± 27.5 and 214 ± 19.8, respectively, showing a stepwise increment in hypomethylation during the SSA/P-cancer sequence. S100P, S100α2, PKP3, and MUC2 were most commonly hypomethylated in SSA/P specimens. The mRNA and protein expression levels of S100P, S100α2, and MUC2 were significantly elevated in SSA/P compared with normal colon tissues, as revealed by RT-PCR and immunohistochemistry, respectively. Among these, mRNA and protein levels were highest for S100P. Knockdown of the S100P gene using a lentiviral shRNA vector in 3-dimensional SSA/P organoids inhibited cell growth by &#x3e;50% (<i>p</i> &#x3c; 0.01). The mean diameter of SSA/P organoids with S100P gene knockdown was significantly smaller compared with control organoids. MassARRAY analysis of DNA hypomethylation in the S100P gene revealed significant hypomethylation at specific CpG sites in intron 1, exon 1, and the 5′-flanking promoter region. <b><i>Conclusion:</i></b> These results suggest that DNA hypomethylation, including S100P hypomethylation, is supposedly associated with the SSA/P-cancer sequence. S100P overexpression via DNA hypomethylation plays an important role in promoting cell growth in the SSA/P-cancer sequence.

EPIGENETIC ALTERATIONS ASSOCIATED WITH PREMATURE OVARIAN FAILURE

2008 ◽  
Vol 31 (4) ◽  
pp. 11
Author(s):  
Manda Ghahremani ◽  
Courtney W Hannah ◽  
Maria Peneherrera ◽  
Karla L Bretherick ◽  
Margo R Fluker ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Premature Ovarian Failure ◽  
Promoter Region ◽  
Gene Promoter ◽  
Ovarian Failure ◽  
P Value ◽  
Epigenetic Alterations ◽  
Methylation Array ◽  
Cpg Sites ◽  
Deficient Mice

Background/Purpose: Premature ovarian failure (POF) affects 1% of women with a largely idiopathic and poorly understood etiology. The objective of this study was to identify specific epigenetic alterations by measuring DNA methylation of gene regulatory regions in women with POF vs. controls. Methods: Blood samples were collected from idiopathic POFpatients (Amenorrhea for at least 3 months and 2 serum FSH levels of > 40mIU/ml obtained > 1 month apart prior to age 40) and control women (CW) (healthy pregnancy after age 37 with out a pregnancy loss). Genomic DNA was extracted from EDTA anticoagulated blood and bisulfite converted for analysis using the Illumina Golden Gate Methylation Panel which measures DNA methylation at 1506 CpG sites in the promoter regions of 807 genes in 10 POF and 12 CW. Candidate genes with altered epigenetic marks between POF and CW at a nominal P-value < 0.05 were identified using a t-testcomparison within the Illumina bead studio software. Genes of interest were further analyzed for quantitative methylation at specific CpG sites using pyrosequencing in 30 POF and 30 CW. Results: Comparison of DNA methylation profiles of our initial POF and CW groups identified several genes with statistically significanthyper- or hypo- methylation in the POF group (P < 0.05), including the Androgen Receptor (AR)promoter region, which was significantly hypermethylated. To further validate these results, DNA methylation of the AR gene promoter was quantified bypryosequencing in a larger group of POF and CW. Pyrosequencing further confirmed a significantly higher DNA methylation of the AR promoter region inPOF vs. CW (P=0.007). Conclusions: This is a novel study identifying epigenetic alterations in POF. The hypermethylation of the AR gene in POF patients may cause decreased level of the AR in these women. This is especially interesting given a recent report of induced POF in AR deficient mice^1. Specific epigenetic markers, as identified by DNA methylation array profiling in blood, may serve as useful biomarkers for POF and other fertility disorders. However, it will need to be determined if these methylation changes are present prior to diagnosis, or are a consequence of menopause itself. Reference: 1.Hiroko S. et al. Premature ovarian failure in androgenreceptor deficient mice. PNAS;103:224-9

lncRNAs as Potential Targets in Small Cell Lung Cancer: MYC -dependent Regulation

2020 ◽  
Vol 20 (17) ◽  
pp. 2074-2081
Author(s):  
Onur Tokgun ◽  
Pervin E. Tokgun ◽  
Kubilay Inci ◽  
Hakan Akca
Keyword(s):  
Lung Cancer ◽  
Stem Cell ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Therapeutic Strategy ◽  
Small Cell ◽  
Small Cell Lung ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Shrna Vector

Background: Small Cell Lung Cancer (SCLC) is a highly aggressive malignancy. MYC family oncogenes are amplified and overexpressed in 20% of SCLCs, showing that MYC oncogenes and MYC regulated genes are strong candidates as therapeutic targets for SCLC. c-MYC plays a fundamental role in cancer stem cell properties and malignant transformation. Several targets have been identified by the activation/repression of MYC. Deregulated expression levels of lncRNAs have also been observed in many cancers. Objective: The aim of the present study is to investigate the lncRNA profiles which depend on MYC expression levels in SCLC. Methods: Firstly, we constructed lentiviral vectors for MYC overexpression/inhibition. MYC expression is suppressed by lentiviral shRNA vector in MYC amplified H82 and N417 cells, and overexpressed by lentiviral inducible overexpression vector in MYC non-amplified H345 cells. LncRNA cDNA is transcribed from total RNA samples, and 91 lncRNAs are evaluated by qRT-PCR. Results: We observed that N417, H82 and H345 cells require MYC for their growth. Besides, MYC is not only found to regulate the expressions of genes related to invasion, stem cell properties, apoptosis and cell cycle (p21, Bcl2, cyclinD1, Sox2, Aldh1a1, and N-Cadherin), but also found to regulate lncRNAs. With this respect, expressions of AK23948, ANRIL, E2F4AS, GAS5, MEG3, H19, L1PA16, SFMBT2, ZEB2NAT, HOTAIR, Sox2OT, PVT1, and BC200 were observed to be in parallel with MYC expression, whereas expressions of Malat1, PTENP1, Neat1, UCA1, SNHG3, and SNHG6 were inversely correlated. Conclusion: Targeting MYC-regulated genes as a therapeutic strategy can be important for SCLC therapy. This study indicated the importance of identifying MYC-regulated lncRNAs and that these can be utilized to develop a therapeutic strategy for SCLC.

scholarly journals A New Multiplatform Model for Outpatient Prenatal and Postpartum Care in a Cohort of COVID-19-Affected Obstetric Patients

2021 ◽  
Vol 18 (10) ◽  
pp. 5144
Author(s):  
Mar Muñoz-Chápuli Gutiérrez ◽  
Ana Durán-Vila ◽  
Javier Ruiz-Labarta ◽  
Pilar Payá-Martínez ◽  
Pilar Pintado Recarte ◽  
...  
Keyword(s):  
Clinical Manifestations ◽  
Specific Treatment ◽  
Rt Pcr ◽  
Good Adherence ◽  
Major Morbidity ◽  
Reference Hospital ◽  
The Mean ◽  
Discharge From Hospital ◽  
Over Time

Spain was one of the epicenters of the first wave of the COVID-19 pandemic. We describe in this article the design and results of a new telephone-and-telematic multiplatform model of systematic prenatal and postpartum follow-up for COVID-19-affected women implemented in a tertiary reference hospital in Madrid. We included patients with RT-PCR-confirmed COVID-19 during pregnancy or delivery from 10 March 2020 to 15 December 2020. We had a total of 211 obstetric patients: 148 (70.1%) were tested at the onset of suspicious clinical manifestations and 62 (29.4%) were tested in the context of routine screening. Of all the patients, 60 women (28.4%) were asymptomatic and 97 (46%) presented mild symptoms. Fifty-one women (24.2%) were admitted to our hospital for specific treatment because of moderate or severe symptoms. We had no missed cases and a good adherence. The mean number of calls per patient was 2.3. We performed 55 in-person visits. We analyzed the complexity of our program over time, showing a two-wave-like pattern. One patient was identified as needing hospitalization and we did not record major morbidity. Telemedicine programs are a strong and reproducible tool to reach to pregnant population affected by COVID-19, to assess its symptoms and severity, and to record for pregnancy-related symptoms both in an outpatient regime and after discharge from hospital.

scholarly journals Comparison of Bite Force in Patients After Treatment of Mandibular Fractures With 3-Dimensional Locking Miniplate and Standard Miniplates

2019 ◽  
Vol 1 (1) ◽  
pp. 7-10
Author(s):  
Gaurav Singh ◽  
Madan Mishra ◽  
Amit Gaur ◽  
Dhritiman Pathak
Keyword(s):  
Treatment Strategies ◽  
Bite Force ◽  
Mandibular Fractures ◽  
Time Intervals ◽  
Experimental Conditions ◽  
3 Dimensional ◽  
The Mean ◽  
Group 2 ◽  
Drilling Conditions ◽  
Group 1

Background: Fractures of the mandible can be studied and described in anatomic terms, functional considerations, treatment strategies, and outcome measures. The performance of any fixation system depends on multiple factors including plate adaptation, screw placement, bone quality, drilling conditions, and postoperative patient compliance. Bite force assesses masticatory muscle function under clinical and experimental conditions. Method: 30 patients with isolated, noncomminuted mandibular fractures were randomly divided into two equal groups. Group 1 patients were treated using 3-dimensional locking miniplates and group 2 patients were treated with standard miniplates. The bite forces were recorded at definite time intervals: preoperatively, and second week, sixth week, third month, and sixth month postoperatively. Result: At 6 weeks postoperative, 3 month postoperative, and 6 month postoperative, the mean bite force was found to be significantly higher among group 1 patients as compared to those in group 2 in all the sites. While at 2 week postoperative, the mean bite force was found to be significantly higher in Group 2 as compared to Group 1 at incisor region. Conclusion: The overall results of the present study show better performance in bite force for the 3-dimensional locking miniplate when compared with standard miniplates.

Quantification of the expression levels of lysosomal cysteine proteinases in purified human osteoclastic cells by competitive RT-PCR

2001 ◽  
Vol 68 (2) ◽  
pp. 109-116 ◽  
Author(s):  
O. Ishibashi ◽  
T. Inui ◽  
Y. Mori ◽  
T. Kurokawa ◽  
T. Kokubo ◽  
...  
Keyword(s):  
Cysteine Proteinases ◽  
Rt Pcr ◽  
Expression Levels

Accuracy of Magnetic Resonance Imaging–Directed Frame-Based Stereotaxis

2011 ◽  
Vol 70 (suppl_1) ◽  
pp. ons114-ons124 ◽  
Author(s):  
Nova B. Thani ◽  
Arul Bala ◽  
Christopher R. P. Lind
Keyword(s):  
Deep Brain Stimulation ◽  
Magnetic Resonance ◽  
Brain Stimulation ◽  
Entry Point ◽  
Mean Deviation ◽  
Guide Tube ◽  
3 Dimensional ◽  
The Mean ◽  
The Brain ◽  
Deep Brain

Abstract BACKGROUND: Accurate placement of a probe to the deep regions of the brain is an important part of neurosurgery. In the modern era, magnetic resonance image (MRI)-based target planning with frame-based stereotaxis is the most common technique. OBJECTIVE: To quantify the inaccuracy in MRI-guided frame-based stereotaxis and to assess the relative contributions of frame movements and MRI distortion. METHODS: The MRI-directed implantable guide-tube technique was used to place carbothane stylettes before implantation of the deep brain stimulation electrodes. The coordinates of target, dural entry point, and other brain landmarks were compared between preoperative and intraoperative MRIs to determine the inaccuracy. RESULTS: The mean 3-dimensional inaccuracy of the stylette at the target was 1.8 mm (95% confidence interval [CI], 1.5-2.1. In deep brain stimulation surgery, the accuracy in the x and y (axial) planes is important; the mean axial inaccuracy was 1.4 mm (95% CI, 1.1-1.8). The maximal mean deviation of the head frame compared with brain over 24.1 ± 1.8 hours was 0.9 mm (95% CI, 0.5-1.1). The mean 3-dimensional inaccuracy of the dural entry point of the stylette was 1.8 mm (95% CI, 1.5-2.1), which is identical to that of the target. CONCLUSION: Stylette positions did deviate from the plan, albeit by 1.4 mm in the axial plane and 1.8 mm in 3-dimensional space. There was no difference between the accuracies at the dura and the target approximately 70 mm deep in the brain, suggesting potential feasibility for accurate planning along the whole trajectory.

scholarly journals miR-142-3p Suppresses Cell Growth by Targeting CDK4 in Colorectal Cancer

2018 ◽  
Vol 51 (4) ◽  
pp. 1969-1981 ◽  
Author(s):  
Xiangyu Zhu ◽  
Si-ping Ma ◽  
Dongxiang Yang ◽  
Yanlong Liu ◽  
Yong-peng Wang ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Cell Growth ◽  
Tumor Cell ◽  
Nude Mice ◽  
Colony Formation ◽  
Tumor Cell Growth ◽  
Rt Pcr ◽  
Tumor Tissues

Background/Aims: Deregulation of microRNAs (miRNAs) has been associated with a variety of cancers, including colorectal cancer (CRC). Here, we investigated anomalous miR-142-3p expression and its possible functional consequences in primary CRC samples. Methods: The expression of miR-142-3p was measured by quantitative RT-PCR in 116 primary CRC tissues and adjacent non-tumor tissues. The effect of miR-142-3p up- or down-regulation in CRC-derived cells was evaluated in vitro by cell viability and colony formation assays and in vivo by growth assays in xenografted nude mice. Results: Using quantitative RT-PCR, we found that miR-142-3p was down-regulated in 78.4 % (91/116) of the primary CRC tissues tested when compared to the adjacent non-tumor tissues. We also found that the miR-142-3p mimic reduced in vitro cell viability and colony formation by inducing cell cycle arrest in CRC-derived cells, and inhibited in vivo tumor cell growth in xenografted nude mice. Inversely, we found that the miR-142-3p inhibitor increased the viability and colony forming capacity of CRC-derived cells and tumor cell growth in xenografted nude mice. In addition, we identified CDK4 as a potential target of miR-142-3p by predictions and dual-luciferase reporter assays. Concordantly, we found that miR-142-3p mimics and inhibitors could decrease and increase CDK4 protein levels in CRC-derived cells, respectively. Conclusion: From our results we conclude that miR-142-3p may act as a tumor suppressor in CRC and may serve as a tool for miRNA-based CRC therapy.

Abstract 11697: Hyperhomocysteinemia Induces CD40 + Monocyte and Inflammatory Monocyte in Chronic Kidney Disease Subjects via DNA Hypomethylation-Related Mechanism

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Jiyeon Yang ◽  
Abhigna C Kolli ◽  
Eric T Choi ◽  
Xiao-Feng Yang ◽  
Hong Wang
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
Blood Cells ◽  
Healthy Donor ◽  
Methylation Status ◽  
Phenotypic Characterization ◽  
Dna Hypomethylation ◽  
Rt Pcr ◽  
Inflammatory Monocyte ◽  
Cd40 Expression

Hyperhomocysteinenia (HHcy) is associated with chronic kidney disease (CKD) which has increased cardiovascular disease (CVD) mortality and mobility. Elevated inflammatory monocyte (inf. MC) is a cellular hallmark of chronic inflammation which contributes to the burden CVD. We previously reported that HHcy induces inf. MC differentiation in mice and DNA hypomethylation in vascular cells. We assesses whether HHcy causes MC differentiation in CKD and the underlying mechanism. Degree of CKD was determined by plasma creatinine from patients with peripheral vascular disease (VD). Estimated glomerular filtration rate was calculated with modification in age, race and gender. (VD, n=13; VD with CKD, n=13; Healthy donor without evidence of VD and CKD, n=13). Blood cells were assessed for phenotypic characterization by flow cytometry. We found that plasma Hcy levels are elevated in VD and CKD. CD14++CD16+ inf. MC are increased in VD subjects and mild HHcy(>15μM). Plasma Hcy levels are positively correlated with inf. MC, CD40, TNF receptor family 5. We identify that CKD patient serum and Hcy (50μM) treatment increased CD40 in purified human blood MC by RT-PCR. Hcy metabolites, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), are increased in CKD subjects. SAM/SAH ratio, an indicator of methylation status, is reduced and is negatively correlated with Hcy, inf. MC, and CD40+ inf. MC. Hcy induced MC-origin inflammatory cytokines IL-6 mRNA in MC isolated from healthy donor by RT-PCR, and potentiated inflammatory cytokine IL-6, TNFα, and IFN[[Unable to Display Character: &#612;]]-induced CD40 expression in cultured PBMCs. Hcy suppressed CD40 transcription and reduced DNA methyltransferase 1 protein levels in cultured human MC. We identified four DNA hypomethylation CpG dinucleotides at p65 and PU.1 transcription factor consensus sequences on CD40 promoter in white blood cells isolated from CKD patients by Bisulfite pyrosequencing. Finally, CD40L levels are positively correlated with plasma Hcy and inf. MC. CD40 ligation by CD40L treatment (0.4μg/l) in peripheral blood mononuclear cells (PBMC) induced inf. MC differentiation. We conclude that HHcy potentiates IFN[[Unable to Display Character: &#612;]]-mediated CD40 expression via CD40 DNA hypomethylation in CKD and promotes CD40-CD40L mediated inf. MC differentiation.

Detecting SARS-CoV-2 by Realtime RT-PCR in clinical samples collected in the North of Vietnam

2021 ◽  
Vol 30 (9) ◽  
pp. 11-17
Author(s):  
Hoang Vu Mai Phuong ◽  
Ung Thi Hong Trang ◽  
Nguyen Vu Son ◽  
Le Thi Thanh ◽  
Nguyen Le Khanh Hang ◽  
...  
Keyword(s):  
Viral Load ◽  
Average Rate ◽  
Clinical Samples ◽  
Viet Nam ◽  
Rt Pcr ◽  
Ct Value ◽  
The North ◽  
Viral Load Data ◽  
The Mean ◽  
High Level

From January to August 2020, Northern Viet Nam faced a COVID-19 outbreak, up to September 2020, there were 1122 confrmed cases of SARS-CoV-2, of which 465 cases were imported from Europe, America and Asia, 657 cases were identifed domestically. A total of 30,686 samples were collected during the SARS-CoV-2 outbreak in Northern Viet Nam and examined by Real-time RT-PCR using primers and probe from Charite - Berlin protocol. This study showed the initial results of SARS-CoV-2 detection and RNA quantitative in positive samples. The positive rate was 0.8%, ranging from 0.4 to 3.5% according to collection sites. Out of 251 positive samples, the mean Ct value was 28 (IQR: 22.3-32; range 14 - 38). The positive samples had a Ct value below 30 was 68.5%, there was no signifcant difference between the Ct value of the group ≤ 30 and > 30. The mean of the RNA copies/µl was 8.4.107, (IQR: 2.29.106 - 1.83.109 RNA copies/µl, range: 1.95.103 – 4.95.1011). In the group of imported COVID-19 cases, the rate of virus at low level was 29%, an average was 56% and at high level was 15%. In the community groups, the viral load data showed that the average rate at low, intermediate and high level were 20%, 63% and 17% respectively. The proportion of high-level viral load may raise an alert to start the quarantine process to reduce the transmission of SARS-CoV-2

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