1280. Insulator Elements Confer High Level, Position Independent, Copy Number Dependent, Uniform Expression of a Linked Transgene Directed by the Erythroid Band 3 (AE1) Promoter in Mice

e15014 Background: Radiotherapy (RT) is one of the main treatments for prostate cancer (PC). The effectiveness of such therapy depends on the initial radioresistance of tumor cells, which is ensured by their certain molecular features, which include the genes copy number variation (CNV). Model experiments on cell cultures (obtained from surgical material) have shown that CNVs have high potential as predictors of RT sensitivity. However, this potential is limited by the high level of invasiveness in obtaining biomaterials. A possible solution to this problem lies in the transition to CNV study in the extracellular DNA (cfDNA) of blood plasma. The aim of the study was to screen predictors of radioresistant PC based on the genes CNV in cfDNA. Methods: The study included 400 patients with diagnosed PC (T2a-3bN0M0, st. II-III), 40 of them after RT had a state of biochemical relapse (RT was performed on a Novalis TX linear accelerator (Varian, USA) (TFDisoeff = 75 Gr), mean time to biochemical relapse 7.5 months). Blood samples were separated into plasma and cell fraction by centrifugation. Isolation of cfDNA from blood plasma was performed using a set of reagents “DNA-Plasma-M” (Russia). Determination of the relative CNV of 13 genes (CDK1, CCND3, CDKN1B, TP53, PTEN, BCL2, XRCC4, BAX, RBBP8, H2AX, BRCA2, RAD50, EP300) was performed using the Real-Time qPCR method. Differences were assessed using the Mann-Whitney test; the Benjamin-Hochberg correction was used to correct multiple comparisons. Results: In the group with biochemical relapse (n = 40), the CNV of genes CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50 was statistically significantly (p < 0.05) higher by 2.0 times, 2.3 times, 2.1 times, 1.4 times, 2.4 times and 2.8 times, respectively, relative to the CNV of these genes in the cfDNA of the group without relapse (n = 360). Conclusions: Thus, it was found that the CNV of 6 genes (CDK1, CDKN1B, RBBP8, XRCC4, BRCA2 and RAD50) may be a potential molecular marker of radiosensitivity of prostate tumors. Based on the obtained data, a low invasive method for determining the prostate tumors sensitivity to RT has been developed.

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High-level expression of porcine muscle adenylate kinase in Escherichia coli: Effects of the copy number of the gene and the translational initiation signals

Journal of Biotechnology ◽

10.1016/0168-1656(94)90176-7 ◽

1994 ◽

Vol 32 (2) ◽

pp. 139-148 ◽

Cited By ~ 12

Author(s):

Takeshi Hibino ◽

Satoru Misawa ◽

Motoaki Wakiyama ◽

Shu Maeda ◽

Kazumori Yazaki ◽

...

Keyword(s):

Escherichia Coli ◽

Adenylate Kinase ◽

Copy Number ◽

High Level Expression ◽

Translational Initiation ◽

Porcine Muscle ◽

High Level

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A cell-specific enhancer far upstream of the mouse tyrosinase gene confers high level and copy number-related expression in transgenic mice.

The EMBO Journal ◽

10.1002/j.1460-2075.1994.tb06607.x ◽

1994 ◽

Vol 13 (13) ◽

pp. 3083-3093 ◽

Cited By ~ 86

Author(s):

R. Ganss ◽

L. Montoliu ◽

A.P. Monaghan ◽

G. Schütz

Keyword(s):

Transgenic Mice ◽

Copy Number ◽

Tyrosinase Gene ◽

A Cell ◽

High Level

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Copy number variation analysis of twin pairs discordant for cleft lip with or without cleft palate

International Journal of Immunopathology and Pharmacology ◽

10.1177/2058738419855873 ◽

2019 ◽

Vol 33 ◽

pp. 205873841985587

Author(s):

Luca Scapoli ◽

Francesco Carinci ◽

Annalisa Palmieri ◽

Francesca Cura ◽

Alessandro Baj ◽

...

Keyword(s):

Cleft Palate ◽

Copy Number ◽

Large Scale ◽

Cleft Lip ◽

Small Sample Size ◽

Copy Number Variants ◽

Monozygotic Twins ◽

Monozygotic Twin ◽

Small Sample ◽

High Level

Non-syndromic cleft lip with or without cleft palate (nsCL/P) is a frequent orofacial malformation. The comparison of concordance rate observed in monozygotic and dizygotic twins supports high level of heritability and a strong genetic component. However, phenotype concordance for orofacial cleft in monozygotic twins is about 50%. The aim of the present investigation was to detect postzygotic events that may account for discordance in monozygotic twins. High-density SNP microarrays hybridization was used to genotype two pairs of monozygotic twins discordant for nsCL/P. Discordant SNP genotypes and copy number variants were analyzed to identify genetic differences responsible of phenotype discrepancy. A number of differences were observed, none involving known nsCL/P candidate genes or genomic regions. Considering the limitation of the study, related to the small sample size and to the large-scale investigation method, the results suggest that the detection of discordant events in other monozygotic twin pairs would be remarkable and warrant further investigations.

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DNA copy number changes in diffuse large B-cell lymphoma--comparative genomic hybridization study

Blood ◽

10.1182/blood.v87.12.5269.bloodjournal87125269 ◽

1996 ◽

Vol 87 (12) ◽

pp. 5269-5278 ◽

Cited By ~ 104

Author(s):

O Monni ◽

H Joensuu ◽

K Franssila ◽

S Knuutila

Keyword(s):

Comparative Genomic Hybridization ◽

Copy Number ◽

B Cell Lymphoma ◽

Comparative Genomic ◽

Genomic Hybridization ◽

Primary Tumors ◽

Large B Cell Lymphoma ◽

Recurrent Tumors ◽

Copy Number Changes ◽

High Level

We studied DNA copy number changes in diffuse large B-cell lymphoma using comparative genomic hybridization analysis on 20 primary tumors and on 12 recurrent tumors excised after chemotherapy or radiotherapy. Twenty-nine (91%) of the cases showed abnormal copy number karyotypes. Chromosomal regions at X (41%), 1q (38%), 7 (31%), 3 (24%), 6p (21%), 11 (21%), 12 (21%), and 18 (21%) were most frequently gained, and the most common losses involved 6q (38%), X (21%), 1p (14%), and 8p (10%). High-level amplifications were observed at 6p23-ter, 10p12–14, 17p1l.2, 18q21-ter, and Xq22-ter, all but 18q appearing only in the recurrent tumors. Gains (median, 2; range, 0 to 10) were more frequent than losses (median, 1; range, 0 to 7; P = .0004). The median number of aberrations found in the recurrent tumors (6.5) was greater than that in the primary tumors (2; P = .01). The copy number changes found in the recurrent tumors were more random than those found in the primary tumors, which were mainly located in the most frequently affected regions. Our findings are in line with those observed using conventional cytogenetic analysis, but especially novel high-level amplifications were detected. Southern blot analysis showed BCL2 amplification, but not translocation t(14;18)(q32;q21), in cases in which a gain at 18q was detected by comparative genomic hybridization, which strongly suggests that, in addition to translocation, gene amplification is another mechanism for the overexpression of the BCL2 protein.

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