Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability

We hypothesized that the role of microbiota in breast cancer relates to its influence on gut lipid metabolism. This was tested in an in vitro model combining MCF-7 and Caco-2 cells. A total of 32 women newly diagnosed for breast cancer before any treatment and 28 healthy women provided their stools....

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Main Authors: Christine Bobin-Dubigeon, Jean-Marie Bard, Trang-Huyen Luu, Françoise Le Vacon, Hassan Nazih
Format: Article
Language:English
Published: MDPI AG 2021-12-01
Series:Nutrients
Subjects:
gut
Online Access:https://www.mdpi.com/2072-6643/13/1/31
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spelling doaj-25324e6050f142999ea8d60cebf178282020-12-25T00:00:17ZengMDPI AGNutrients2072-66432021-12-0113313110.3390/nu13010031Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell ViabilityChristine Bobin-Dubigeon0Jean-Marie Bard1Trang-Huyen Luu2Françoise Le Vacon3Hassan Nazih4EA 2160—IUML FR3473 CNRS, Université de Nantes, 44035 Nantes, FranceEA 2160—IUML FR3473 CNRS, Université de Nantes, 44035 Nantes, FranceEA 2160—IUML FR3473 CNRS, Université de Nantes, 44035 Nantes, FranceBiofortis, Mérieux Nutrisciences, 44800 Saint-Herblain, FranceEA 2160—IUML FR3473 CNRS, Université de Nantes, 44035 Nantes, FranceWe hypothesized that the role of microbiota in breast cancer relates to its influence on gut lipid metabolism. This was tested in an in vitro model combining MCF-7 and Caco-2 cells. A total of 32 women newly diagnosed for breast cancer before any treatment and 28 healthy women provided their stools. Bacterial DNA was amplified by qPCR targeting 16s rRNA specific to Bacteroidetes and Firmicutes phyla, <i>Lactobacillales</i> sp., <i>Clostridium</i> cluster IV, <i>Faecalibacterium prausnitzii</i>, <i>Clostridium</i> cluster XIVa, <i>Roseburia intestinalis</i>, <i>Blautia</i> sp., <i>Lactonifactor longoviformis</i>, <i>Bifidobacterium</i> sp., Coriobacteriaceae, <i>Eggertella lenta</i>, <i>Escherichia,</i> and <i>Shigella</i>. Fecal waters (FW) were quantified for short chain fatty acids (SCFA). Caco-2 cells grown on filter inserts were incubated apically with 10% FW for 24 h, and <i>LXR</i>, apolipoproteins <i>AIV,</i> and <i>E</i> gene expression were estimated by real time (RT) qPCR. Then, MCF-7 cells were incubated with the whole basolateral medium for 24 h, and their viability was estimated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) test. Regression models were used to determine the correlation between MCF-7 viability and bacteria relative abundance, Caco-2 cells lipid metabolism gene expression and stool composition, as well as microbiota composition and short chain fatty acids. Logistic regression models established disease odds ratios (OR) for MCF-7 viability and Caco-2 gene expression. The OR of MCF-7 viability was 1.05 (1.01–1.10) (OR (5th–95th), <i>p</i> = 0.04), while that of <i>apo AIV</i> gene expression was 0.63 (0.39–1.01), <i>p</i> = 0.055). Viability correlated with % <i>Bifidobacterium</i> sp. (21.18 ± 7.66, <i>p</i> = 0.008) and valerate (−2.849 ± 1.048, <i>p</i> = 0.009) (β ± s.d.). This study suggests that microbiota interacts with intestine cell lipid metabolism. Since these metabolites can reach breast cells by systemic circulation, we hypothesized that they may influence cancer disease.https://www.mdpi.com/2072-6643/13/1/31microbiotabreast cancergutlipid metabolism
collection DOAJ
language English
format Article
sources DOAJ
author Christine Bobin-Dubigeon
Jean-Marie Bard
Trang-Huyen Luu
Françoise Le Vacon
Hassan Nazih
spellingShingle Christine Bobin-Dubigeon
Jean-Marie Bard
Trang-Huyen Luu
Françoise Le Vacon
Hassan Nazih
Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
Nutrients
microbiota
breast cancer
gut
lipid metabolism
author_facet Christine Bobin-Dubigeon
Jean-Marie Bard
Trang-Huyen Luu
Françoise Le Vacon
Hassan Nazih
author_sort Christine Bobin-Dubigeon
title Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
title_short Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
title_full Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
title_fullStr Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
title_full_unstemmed Basolateral Secretion from Caco-2 Cells Pretreated with Fecal Waters from Breast Cancer Patients Affects MCF7 Cell Viability
title_sort basolateral secretion from caco-2 cells pretreated with fecal waters from breast cancer patients affects mcf7 cell viability
publisher MDPI AG
series Nutrients
issn 2072-6643
publishDate 2021-12-01
description We hypothesized that the role of microbiota in breast cancer relates to its influence on gut lipid metabolism. This was tested in an in vitro model combining MCF-7 and Caco-2 cells. A total of 32 women newly diagnosed for breast cancer before any treatment and 28 healthy women provided their stools. Bacterial DNA was amplified by qPCR targeting 16s rRNA specific to Bacteroidetes and Firmicutes phyla, <i>Lactobacillales</i> sp., <i>Clostridium</i> cluster IV, <i>Faecalibacterium prausnitzii</i>, <i>Clostridium</i> cluster XIVa, <i>Roseburia intestinalis</i>, <i>Blautia</i> sp., <i>Lactonifactor longoviformis</i>, <i>Bifidobacterium</i> sp., Coriobacteriaceae, <i>Eggertella lenta</i>, <i>Escherichia,</i> and <i>Shigella</i>. Fecal waters (FW) were quantified for short chain fatty acids (SCFA). Caco-2 cells grown on filter inserts were incubated apically with 10% FW for 24 h, and <i>LXR</i>, apolipoproteins <i>AIV,</i> and <i>E</i> gene expression were estimated by real time (RT) qPCR. Then, MCF-7 cells were incubated with the whole basolateral medium for 24 h, and their viability was estimated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) test. Regression models were used to determine the correlation between MCF-7 viability and bacteria relative abundance, Caco-2 cells lipid metabolism gene expression and stool composition, as well as microbiota composition and short chain fatty acids. Logistic regression models established disease odds ratios (OR) for MCF-7 viability and Caco-2 gene expression. The OR of MCF-7 viability was 1.05 (1.01–1.10) (OR (5th–95th), <i>p</i> = 0.04), while that of <i>apo AIV</i> gene expression was 0.63 (0.39–1.01), <i>p</i> = 0.055). Viability correlated with % <i>Bifidobacterium</i> sp. (21.18 ± 7.66, <i>p</i> = 0.008) and valerate (−2.849 ± 1.048, <i>p</i> = 0.009) (β ± s.d.). This study suggests that microbiota interacts with intestine cell lipid metabolism. Since these metabolites can reach breast cells by systemic circulation, we hypothesized that they may influence cancer disease.
topic microbiota
breast cancer
gut
lipid metabolism
url https://www.mdpi.com/2072-6643/13/1/31
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