KRAS mutation analysis of single circulating tumor cells from patients with metastatic colorectal cancer

Abstract

Background

The molecular profiles of tumors may inform the selection of appropriate targeted therapies. Circulating tumor cells (CTCs) reflect the real-time status of tumor genotypes. CTCs exhibit high genetic heterogeneity within a patient; accordingly, the analysis of individual CTCs, including their heterogeneity, may enable more precise treatments. We analyzed KRAS mutations in single CTCs from patients with metastatic colorectal cancer (mCRC) using a new single-cell picking system.

Methods

Blood samples were obtained from 61 patients with mCRC. CTCs were enriched and fluorescently labeled using the CellSearch® System. They were recovered using the single-cell picking system based on the fluorescence intensity of marker dyes. Single CTCs and tumor tissue samples were examined for mutations in codons 12 and 13 of the KRAS gene.

Results

CTCs were detected in 27 of 61 patients with mCRC. We isolated at least two CTCs from 15 of 27 patients. KRAS genotype was evaluated in a total of 284 CTCs from 11 patients, and 15 cells with mutations were identified in four patients. In 10 of 11 patients, the KRAS status was the same in the primary tumor and CTCs. In one patient, the KRAS status was discordant between the primary tumor and CTCs. In two patients, different KRAS mutations were found among individual CTCs.

Conclusions

We successfully isolated single CTCs and detected KRAS mutations in individual cells from clinical samples using a novel application of single-cell isolation system. Using the system, we detected CTC heterozygosity and heterogeneity in KRAS status among CTCs within a patient and between CTCs and tumor tissues.

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Soluble MICA is elevated in pancreatic cancer: results from a population based case-control study

Abstract

Objectives: Pancreatic cancer is diagnosed at a late stage and has one of the highest cancer mortality rates in the U.S., creating an urgent need for novel early detection tools. A candidate biomarker for use in early detection is the soluble MHC class I-related chain A (s-MICA) ligand, which pancreatic tumors shed to escape immune detection. The objective of this study was to define the association between s-MICA levels and pancreatic cancer, in a population-based case-control study.

Methods: S-MICA was measured in 163 pancreatic cancer cases and 542 controls. Unconditional logistic regression was used to calculate odds ratio (OR) for pancreatic cancer, and 95% confidence intervals (CI). Results: There was a positive association between increasing s-MICA levels and pancreatic cancer: compared to the lowest tertile, the ORs for pancreatic cancer were 1.25 (95% CI: 0.75 - 2.07) and 2.10 (95% CI: 1.29 -3.42) in the second and highest tertiles, respectively (p-trend=0.02).

Conclusions: Our study supports previous work demonstrating a positive association between plasma s-MICA levels and pancreatic cancer. This article is protected by copyright. All rights reserved

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Glyoxalase 2 drives tumorigenesis in human prostate cells in a mechanism involving androgen receptor and p53-p21 axis

Abstract

Glyoxalase 2 (Glo2), a metabolic enzyme, is overexpressed in some human cancers which suggests this enzyme may play a role in human tumorigenesis. In prostate cancer (PCa) the role of Glo2 has been scarcely investigated and there are no studies addressing a causative involvement of this protein in this neoplasia. Here we examined the immunohistochemical profile of Glo2 in human PCa and benign adjacent tissues and investigated Glo2 involvement in PCa development in human prostate cell lines. PCa and matched adjacent normal tissues were obtained from paraffin sections of primary PCa from 20 patients who had undergone radical prostatectomy. Histopathological diagnosis was confirmed for each sample. Glo2 expression analysis was performed by immunohistochemistry in prostate tissues, and by qRT-PCR and immunoblotting in prostate cell lines. The causative and mechanistic role of Glo2 in prostate tumorigenesis was demonstrated by Glo2 ectopic expression/silencing and employing specific activators/inhibitors. Our results showed that Glo2 was selectively expressed in PCa but not in the luminal compartment of the adjacent benign epithelium consistently in all the examined 20 cases. Glo2 expression in PCa was dependent on androgen receptor (AR) and was aimed at stimulating cell proliferation and eluding apoptosis through a mechanism involving the p53-p21 axis. Glo2 was intensely expressed in the basal cells of benign glands but was not involved in PCa genesis. Our results demonstrate for the first time that Glo2 drives prostate tumorigenesis and suggest that it may represent a novel adjuvant marker in the pathological diagnosis of early PCa. This article is protected by copyright. All rights reserved

http://ift.tt/2qHbwCg

KRAS mutation analysis of single circulating tumor cells from patients with metastatic colorectal cancer

Abstract

Background

The molecular profiles of tumors may inform the selection of appropriate targeted therapies. Circulating tumor cells (CTCs) reflect the real-time status of tumor genotypes. CTCs exhibit high genetic heterogeneity within a patient; accordingly, the analysis of individual CTCs, including their heterogeneity, may enable more precise treatments. We analyzed KRAS mutations in single CTCs from patients with metastatic colorectal cancer (mCRC) using a new single-cell picking system.

Methods

Blood samples were obtained from 61 patients with mCRC. CTCs were enriched and fluorescently labeled using the CellSearch® System. They were recovered using the single-cell picking system based on the fluorescence intensity of marker dyes. Single CTCs and tumor tissue samples were examined for mutations in codons 12 and 13 of the KRAS gene.

Results

CTCs were detected in 27 of 61 patients with mCRC. We isolated at least two CTCs from 15 of 27 patients. KRAS genotype was evaluated in a total of 284 CTCs from 11 patients, and 15 cells with mutations were identified in four patients. In 10 of 11 patients, the KRAS status was the same in the primary tumor and CTCs. In one patient, the KRAS status was discordant between the primary tumor and CTCs. In two patients, different KRAS mutations were found among individual CTCs.

Conclusions

We successfully isolated single CTCs and detected KRAS mutations in individual cells from clinical samples using a novel application of single-cell isolation system. Using the system, we detected CTC heterozygosity and heterogeneity in KRAS status among CTCs within a patient and between CTCs and tumor tissues.

from Cancer via ola Kala on Inoreader http://ift.tt/2qBRdXF
via IFTTT

Soluble MICA is elevated in pancreatic cancer: results from a population based case-control study

Abstract

Objectives: Pancreatic cancer is diagnosed at a late stage and has one of the highest cancer mortality rates in the U.S., creating an urgent need for novel early detection tools. A candidate biomarker for use in early detection is the soluble MHC class I-related chain A (s-MICA) ligand, which pancreatic tumors shed to escape immune detection. The objective of this study was to define the association between s-MICA levels and pancreatic cancer, in a population-based case-control study.

Methods: S-MICA was measured in 163 pancreatic cancer cases and 542 controls. Unconditional logistic regression was used to calculate odds ratio (OR) for pancreatic cancer, and 95% confidence intervals (CI). Results: There was a positive association between increasing s-MICA levels and pancreatic cancer: compared to the lowest tertile, the ORs for pancreatic cancer were 1.25 (95% CI: 0.75 - 2.07) and 2.10 (95% CI: 1.29 -3.42) in the second and highest tertiles, respectively (p-trend=0.02).

Conclusions: Our study supports previous work demonstrating a positive association between plasma s-MICA levels and pancreatic cancer. This article is protected by copyright. All rights reserved

from Cancer via ola Kala on Inoreader http://ift.tt/2qukqqj
via IFTTT

Glyoxalase 2 drives tumorigenesis in human prostate cells in a mechanism involving androgen receptor and p53-p21 axis

Abstract

Glyoxalase 2 (Glo2), a metabolic enzyme, is overexpressed in some human cancers which suggests this enzyme may play a role in human tumorigenesis. In prostate cancer (PCa) the role of Glo2 has been scarcely investigated and there are no studies addressing a causative involvement of this protein in this neoplasia. Here we examined the immunohistochemical profile of Glo2 in human PCa and benign adjacent tissues and investigated Glo2 involvement in PCa development in human prostate cell lines. PCa and matched adjacent normal tissues were obtained from paraffin sections of primary PCa from 20 patients who had undergone radical prostatectomy. Histopathological diagnosis was confirmed for each sample. Glo2 expression analysis was performed by immunohistochemistry in prostate tissues, and by qRT-PCR and immunoblotting in prostate cell lines. The causative and mechanistic role of Glo2 in prostate tumorigenesis was demonstrated by Glo2 ectopic expression/silencing and employing specific activators/inhibitors. Our results showed that Glo2 was selectively expressed in PCa but not in the luminal compartment of the adjacent benign epithelium consistently in all the examined 20 cases. Glo2 expression in PCa was dependent on androgen receptor (AR) and was aimed at stimulating cell proliferation and eluding apoptosis through a mechanism involving the p53-p21 axis. Glo2 was intensely expressed in the basal cells of benign glands but was not involved in PCa genesis. Our results demonstrate for the first time that Glo2 drives prostate tumorigenesis and suggest that it may represent a novel adjuvant marker in the pathological diagnosis of early PCa. This article is protected by copyright. All rights reserved

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via IFTTT

Cure rate of postpartum endometritis after different treatments in high produce dairy cows

Abstract

The main objective of this study was to compare the common treatment methods of clinical endometritis (CE) including injection of PGF2a and intrauterine infusion of cephapirin benzathine (CB) with intrauterine infusion of 50% dextrose solution (DEX) and intrauterine infusion of liquid paraffin (LP) in dairy cows. To this end, 188 lactating Holstein cows with clinical signs of endometritis were randomly assigned into five treatment groups and cytological smear was prepared. In group 1 (n = 37), cows were treated with intrauterine infusion of CB. In the second group (n = 53), the treatment protocol included intramuscular injection of 500-mg cloprostenol sodium in cows with corpus luteum (CL) in their ovaries. In group 3 (n = 43), cows were treated with intrauterine infusion of DEX solution. In group 4 (n = 35), cows were treated with intrauterine infusion of LP solution and 20 untreated cows were assigned into a control group (group 5). Clinical treatment rates were 62.2, 66, 60.5, 54.3, and 50% in CB, PG, DEX, LP, and control groups, respectively (P ≥ 0.05). In the cytological assessment, there was no significant difference in reduction in the neutrophil's number in the second examination after treatment (P ≥ 0.05). Percentage of cows with clean vaginal discharge in the second examination is higher in cows with CL than cows without CL in all five groups, but this difference was not significant (P ≥ 0.05). According to the result of this study, future large studies are required for assessment of DEX and LP as alternative therapies for the treatment of CE instead of antibiotics.

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