Bcl-2 is closely correlated with favorable prognostic factors and inversely associated with p53 protein accumulation in endometrial carcinomas: immunohistochemical and polymerase chain reaction/loss of heterozygosity findings

1997 ◽  
Vol 123 (8) ◽  
pp. 429-434 ◽  
Author(s):  
Makoto Saegusa ◽  
Isao Okayasu
Keyword(s):  
Polymerase Chain Reaction ◽  
Prognostic Factors ◽  
Loss Of Heterozygosity ◽  
P53 Protein ◽  
Protein Accumulation ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Endometrial Carcinomas

Novel Method for Simultaneous Analysis of p53 and K-ras Mutations and p53 Protein Expression in Single Histologic Sections

2001 ◽  
Vol 125 (3) ◽  
pp. 347-352
Author(s):  
Kazuya Yamashita ◽  
Tsutomu Yoshida ◽  
Hiroshi Shinoda ◽  
Isao Okayasu
Keyword(s):  
Polymerase Chain Reaction ◽  
Protein Expression ◽  
P53 Protein ◽  
Single Strand Conformation Polymorphism ◽  
Antigen Retrieval ◽  
Cancer Tissue ◽  
Polymorphism Analysis ◽  
Chain Reaction ◽  
Novel Method ◽  
Polymerase Chain

Abstract Background and Objective.—Abnormal protein expression and gene mutation should be examined on exactly identified lesions. To perform simultaneous analyses of oncogene or tumor suppressor gene mutations and related protein expression in single histologic sections, we have developed a novel method using an antigen-retrieval solution for a polymerase chain reaction template before immunohistochemical staining. Methods.—Using 20 cases of sporadic colorectal carcinoma, several kinds of antigen-retrieval solutions were tested after heating rehydrated, 4-μm-thick, formalin-fixed, paraffin-embedded histologic sections at 96°C for 20 minutes. Polymerase chain reaction–single-strand conformation polymorphism analysis was conducted for p53 (exons 5 through 9) and K-ras (exons 1 and 2), and the histologic sections were then immunostained with monoclonal antibody against p53. Results.—DNA analysis of antigen-retrieval solutions was possible in all 20 cases and revealed completely consistent results (100%) with fresh cancer tissue and microdissected cancer tissue of paraffin-embedded histologic sections. With this method, K-ras mutations were positive in 10 of 20 cases (exon 1 in 9 cases and exon 2 in 1 case) and p53 mutations were positive in 9 of 20 cases (exon 5 in 4 cases, exon 6 in 1, exon 7 in 3, and exon 8 in 1 case), with 8 of the 9 p53 mutation cases showing diffuse p53 protein expression on immunostaining. Base alterations of all abnormal conformers were confirmed with direct sequencing. For polymerase chain reaction–single-strand conformation polymorphism analysis, sodium citrate buffer (pH 6.0) was found to be the optimal antigen-retrieval solution. Conclusions.—This newly developed method can be used for routine immunostaining and genetic analysis with single histologic sections.

scholarly journals Myeloid but not lymphoid cells carry the 5q deletion: polymerase chain reaction analysis of loss of heterozygosity using mini-repeat sequences on highly purified cell fractions

Blood ◽  
1993 ◽  
Vol 81 (7) ◽  
pp. 1849-1854 ◽  
Author(s):  
MJ Kroef ◽  
WE Fibbe ◽  
R Mout ◽  
RP Jansen ◽  
HL Haak ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Loss Of Heterozygosity ◽  
Methylation Status ◽  
Pcr Amplification ◽  
Lymphoid Cells ◽  
Allelic Loss ◽  
Chain Reaction ◽  
Ca Repeat ◽  
Repeat Sequences ◽  
Polymerase Chain

Interstitial deletions of the long arm of chromosome 5 are among the most characteristic abnormalities observed in myeloid disorders. To assess the lineage involvement of peripheral blood cells from patients with a 5q--anomaly, purified neutrophils, monocytes, T lymphocytes, and B lymphocytes were analyzed for loss of heterozygosity using six different highly polymorphic mininucleotide and dinucleotide (CA) repeat sequences from the 5q31 to 5q33 region. Ten patients were screened by polymerase chain reaction (PCR) amplification and proved to be informative for at least one marker. Six patients showed a complete or partial disappearance of an allele in myeloid cells, whereas cells of lymphoid lineages exhibited full heterozygosity. The other patients displayed no allelic loss, indicating that the informative markers were located outside the deleted chromosomal segments. In addition, three female patients who were also polymorphic for the BstXI site in the PGK- 1 gene were analyzed for the methylation status of this gene. Clonality of hematopoiesis, as determined by non-random X-chromosome inactivation, followed the same cell pattern as the 5q-specific allelic losses. In conclusion, using tumor-specific and clonal markers, we have demonstrated that the 5q- anomaly is restricted to cells of myeloid origin, leaving lymphoid cells unaffected.

scholarly journals Analysis of BRCA2 loss of heterozygosity in tumor tissue using droplet digital polymerase chain reaction

2014 ◽  
Vol 45 (7) ◽  
pp. 1546-1550 ◽  
Author(s):  
Rory L. Cochran ◽  
Karen Cravero ◽  
David Chu ◽  
Bracha Erlanger ◽  
Patricia Valda Toro ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Loss Of Heterozygosity ◽  
Tumor Tissue ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction
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