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Histochemical and immunohistochemical evidence of tumor heterogeneity in colorectal cancer.

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TLDR
The study demonstrated the intratumoral and intertumoral heterogeneity, expressed at phenotypic level in colorectal adenocarcinomas, and found a significant positive correlation between KRAS protein expression and BCL2 and TP53 expression.
Abstract
Introduction Intratumoral heterogeneity implies the existence of differences between tumor cells, which can best be shown by histochemical and immunohistochemical techniques. The histological study is a mandatory step in any research aimed at characterizing tumor heterogeneity. Immunohistochemistry (IHC) also plays an important role in the differentiation of tumor types, assessing aggressiveness. Materials and methods Investigated group consisted of 50 patients with colorectal adenocarcinoma, for each were recorded clinicopathological data and harvested samples intraoperatively, which were included in paraffin blocks. We perform Hematoxylin-Eosin staining for histological grade and other indices. IHC study used Avidin-Biotin-Peroxidase (ABC), with the markers: CK7, CK20, MUC1, MUC2, Ki-67, PCNA, p53, KRAS, BCL2, PTEN, EGFR. The resulting data were analyzed by statistical methods. Results Most of colorectal adenocarcinoma studied had no special histological features and had G2 grade. IHC detected in most cases the CK20+÷CK7- phenotype (78%) and MUC1 (74%) protein expression. The proliferation markers (Ki-67 and PCNA) were present in all tumor mass with a variable index, which shows high intratumoral heterogeneity, but p53 and KRAS were distributed more uniformly, showing low intratumoral heterogeneity. PTEN was expressed nuclearly in 86% of the cases and EGFR in 42%. Conclusions The expression profiles of cytokeratins and mucins in the colorectal adenocarcinomas are useful in defining tumor phenotypes with different prognosis and therapy. We found a significant positive correlation between KRAS protein expression and BCL2 and TP53 expression. The study demonstrated the intratumoral and intertumoral heterogeneity, expressed at phenotypic level.

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References
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Ki-67 is a PP1-interacting protein that organises the mitotic chromosome periphery

TL;DR: In this paper, the authors report that the nucleolar protein Ki-67 is required for the assembly of the perichromosomal compartment in human cells, which is a cell-cycle regulated protein phosphatase 1-binding protein that is involved in phospho-regulation of B23/nucleophosmin.
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Antibody Elution Method for Multiple Immunohistochemistry on Primary Antibodies Raised in the Same Species and of the Same Subtype

TL;DR: This work presents a procedure that elutes the antibodies after a first round of immunolabeling, which, in combination with precipitation-based detection systems, allows multiple IHC rounds even for primary antibodies raised in the same species and IgG isotype.
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The Ki-67 protein interacts with members of the heterochromatin protein 1 (HP1) family: a potential role in the regulation of higher-order chromatin structure.

TL;DR: It is shown that a C‐terminal domain of pKi‐67 (Kon21) is able to bind to all three members of the mammalian heterochromatin protein 1 (HP1) family in vitro and in vivo, and this interaction can be manipulated in living cells, as evidenced by ectopic expression of GFP‐tagged HP1 proteins in HeLa cells, which results in a dramatic relocalization of endogenous pKI‐67.
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Cytokeratin 7 and cytokeratin 20 expression in colorectal adenocarcinomas.

TL;DR: CK7 positivity or CK20 negativity does not rule out a colorectal origin of metastatic carcinoma, and CK7 expression was more common in tumors with lymph node metastasis than in non-metastatic tumors.
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