Comparison of BCR/ABL1 mRNA levels by quantitative real-time PCR in peripheral blood and bone marrow specimens of patients with chronic myelogenous leukemia

Abstract Background Wilms’ tumor, also called nephroblastoma, is the most common pediatric renal malignancy. The pathogenesis of Wilms’ tumor has been attributed to several genetic and epigenetic factors. However, the most pervasive internal mRNA modification that affects almost every process of RNA metabolism, RNA N6-Methyladenosine (m6A) methylation, has not been characterized in Wilms’ tumor. Methods Wilms’ tumor (WT) and adjacent non-cancerous (NC) tissue samples were obtained from 23 children with nephroblastoma, and the global m6A levels were measured by mass spectrometry. Analyses by m6A-mRNA epitranscriptomic microarray and mRNA microarray were performed, and m6A-related mRNAs were validated by quantitative real-time PCR for input and m6A-immunoprecipitated RNA samples from WT and NC tissues. Gene ontology analysis and KEGG pathway analysis were performed for differentially expressed genes, and expression of RNA methylation-related factors was measured by quantitative real-time PCR. Results The total m6A methylation levels in total RNA of WT samples and NC samples were (0.21 ± 0.01)% and (0.22 ± 0.01)%, respectively, with no statistically significant difference. Fifty-nine transcripts were differentially m6A-methylated between the WT and NC groups, which showed distinct m6A modification patterns. Gene ontology analysis indicated that m6A-modified genes were enriched in cancer-associated pathways, including the mTOR pathway, and conjoint analysis of the unique methylation and gene expression patterns in WT samples suggested an association with metabolic pathways.The mRNA levels of the m6A-related “reader” genes, YTHDF1, YTHDF2 and IGF2BP3, were statistically higher in WT samples than in NC samples. Conclusion This is the first study to determine the m6A modification profiles in Wilms’ tumor. Our data provide novel information regarding patterns of m6A modification that correlate with carcinogenesis in Wilms’ tumor.

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Congenital Juvenile Chronic Myelogenous Leukemia: Case Report and Review

PEDIATRICS ◽

10.1542/peds.73.3.324 ◽

1984 ◽

Vol 73 (3) ◽

pp. 324-326

Author(s):

Reese H. Clark ◽

Leslie L. Taylor ◽

Robert J. Wells

Keyword(s):

Bone Marrow ◽

Case Report ◽

Chronic Myelogenous Leukemia ◽

Peripheral Blood ◽

Blast Crisis ◽

Myelogenous Leukemia ◽

Juvenile Form

The case of a patient with ecchymosis, hepatomegaly, leukocytosis, thrombocytopenia, and anemia at birth is presented. Throughout his course, thrombocytopenia, anemia, and leukocytosis without a marked increase in the number of blast forms in either peripheral blood or bone marrow persisted until the patient developed a blast crisis shortly before his death at age 4 months. This patient is the youngest reported to have the juvenile form of chronic myelogenous leukemia and the first that in the present era can be considered congenital in origin.

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Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

Archives of Pathology & Laboratory Medicine ◽

10.5858/2008-132-813-isobbm ◽

2008 ◽

Vol 132 (5) ◽

pp. 813-819

Author(s):

Xiaohong Han ◽

Jeffrey L. Jorgensen ◽

Archana Brahmandam ◽

Ellen Schlette ◽

Yang O. Huh ◽

...

Keyword(s):

Bone Marrow ◽

Flow Cytometry ◽

Chronic Myelogenous Leukemia ◽

Peripheral Blood ◽

Myelogenous Leukemia ◽

Multiparameter Flow Cytometry ◽

Color Flow ◽

Aberrant Expression ◽

Flow Cytometric ◽

Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

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Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Journal of Visualized Experiments ◽

10.3791/50193 ◽

2013 ◽

Cited By ~ 2

Author(s):

Ningfei An ◽

Yubin Kang

Keyword(s):

Bone Marrow ◽

Bone Marrow Transplantation ◽

Real Time ◽

Real Time Pcr ◽

Marrow Transplantation ◽

Donor Cell ◽

Quantitative Real Time Pcr ◽

Murine Bone Marrow ◽

Cell Engraftment ◽

Transplantation Model

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Cloning, Characterization, and Expression Features of Chicken CDS2 Splicing Variants

10.21203/rs.3.rs-80587/v1 ◽

2020 ◽

Author(s):

Yuanyuan Xu ◽

Shuping Zhang ◽

Yujun Guo ◽

Wen Chen ◽

Yanqun Huang

Keyword(s):

Adipose Tissue ◽

Real Time ◽

Real Time Pcr ◽

Expression Patterns ◽

Mrna Levels ◽

Exogenous Insulin ◽

Quantitative Real Time Pcr ◽

Temporal Expression ◽

Spatio Temporal ◽

The Brain

Abstract Background: The CDS gene encodes the CDP-diacylglycerol synthase enzyme that catalyzes the formation of CDP-diacylglycerol (CDP-DAG) from phosphatidic acid. At present, there are no reports of CDS2 in birds. Here, we identified chicken CDS2 transcripts by combining conventional RT- PCR amplification, 5' RACE (Fig. 1A), and 3' RACE, explored the spatio-temporal expression profiles of total CDS2 and the longest transcript variant CDS2-4, and investigated the effect of exogenous insulin on total the mRNA level of CDS2 by quantitative real-time PCR. Results: Four transcripts of chicken CDS2 (CDS2-1, -2, -3, and -4) were identified, which were alternatively spliced at the 3′-untranslated region (UTR). CDS2 was widely expressed in all tissues examined and the longest variant CDS2-4 was the major transcript. Both total CDS2 and CDS2-4 were prominently expressed in adipose tissue and the heart, and exhibited low expression in the liver and pectoralis of 49 day-old chickens. Quantitative real-time PCR revealed that total CDS2 and CDS2-4 had different spatio-temporal expression patterns in chicken. Total CDS2 exhibited a similar temporal expression tendency with a high level in the later period of incubation (embryonic day 19 [E19] or 1-day-old) in the brain, liver, and pectoralis. While CDS2-4 presented a distinct temporal expression pattern in these tissues, CDS2-4 levels peaked at 21 days in the brain and pectoralis, while liver CDS2-4 mRNA levels were highest at the early stage of hatching (E10). Total CDS2 (P < 0.001) and CDS2-4 (P = 0.0090) mRNA levels in the liver were differentially regulated throughout development of the chicken. Exogenous insulin significantly downregulated the level of total CDS2 at 240 min in the pectoralis of Silky chickens (P < 0.01). Total CDS2 levels in the liver of Silky chickens were higher than that of the broiler in the basal state and after insulin stimulation. Conclusion: Chicken CDS2 has multiple transcripts with variation at the 3′-UTR, which was prominently expressed in adipose tissue. Total CDS2 and CDS2-4 presented distinct spatio-temporal expression patterns, and they were differentially regulated with age in liver. Insulin could regulate chicken CDS2 levels in a breed- and tissue-specific manner.

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Discovery of candidate gene expression signatures in peripheral blood for the screening of cervical cancer

Biomarkers in Medicine ◽

10.2217/bmm-2019-0247 ◽

2020 ◽

Vol 14 (2) ◽

pp. 109-118 ◽

Cited By ~ 2

Author(s):

Qiuling Ma ◽

Yong Shao ◽

Wei Chen ◽

Cheng Quan ◽

Yanhui Zhu ◽

...

Keyword(s):

Gene Expression ◽

Cervical Cancer ◽

Real Time ◽

Real Time Pcr ◽

Peripheral Blood ◽

Area Under The Curve ◽

Intraepithelial Neoplasia ◽

Sequencing Analysis ◽

Quantitative Real Time Pcr ◽

Cervical Lesions

Aim: To investigate whether cervical cancer (CC) and cervical intraepithelial neoplasia (CIN) can be screened by analyzing gene expression profiling of peripheral blood. Methods: RNA-sequencing analysis of blood was performed on 11 CC patients, 21 CIN patients and 19 healthy controls (H). Fifty-nine genes were validated by quantitative real-time PCR using blood samples from 46 H, 83 CC and 32 CIN patients. Results: There were significant differences in the expression levels of six genes between CC and H, five genes between CIN and H and four genes between CC and CIN (p < 0.05). Four genes discriminated cervical lesions from H with a sensitivity of 82.61%, a specificity of 87.83% and an area under the curve of 0.8981. Three genes discriminated CC from CIN with a sensitivity of 53.13%, a specificity of 96.39% and an area under the curve of 0.7786. Conclusion: Our findings provided a promising noninvasive quantitative real-time PCR diagnostic assay of CC and CIN.

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Detection by enzymatic amplification of bcr-abl mRNA in peripheral blood and bone marrow cells of patients with chronic myelogenous leukemia

Blood ◽

10.1182/blood.v73.6.1735.1735 ◽

1989 ◽

Vol 73 (6) ◽

pp. 1735-1741 ◽

Cited By ~ 44

Author(s):

W Lange ◽

DS Snyder ◽

R Castro ◽

JJ Rossi ◽

KG Blume

Keyword(s):

Bone Marrow ◽

Chronic Myelogenous Leukemia ◽

Peripheral Blood ◽

Bone Marrow Cells ◽

Philadelphia Chromosome ◽

Chromosome 9 ◽

Myelogenous Leukemia ◽

Enzymatic Amplification ◽

Polymerase Chain ◽

Marrow Cells

Abstract The Philadelphia chromosome of chronic myelogenous leukemia (CML) patients is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. A new bcr- abl mRNA is expressed in these cases. We have developed a modified polymerase chain reaction (PCR) for the detection of this mRNA. The method is extremely sensitive, reliable, and relatively fast. The analysis of peripheral blood or bone marrow cells from CML patients treated with chemotherapy shows that the two possible mRNAs are expressed in various combinations. Our results show that even after myeloablative therapy for bone marrow transplantation bcr-abl mRNAs are still expressed. Further studies, however, are necessary to determine the clinical relevance of a small number of persisting cells expressing the bcr-abl mRNA.

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