Oleic Acid Uptake Reveals the Rescued Enterocyte Phenotype of Colon Cancer Caco-2 by HT29-MTX Cells in Co-Culture Mode
Abstract
:1. Introduction
2. Results
2.1. Bio-Informatic Dataset Analyses Point out the Role of Fatty Acids in Both Cancer and Intestinal Phenotypes
2.2. Real-Time Analysis Reveals the Modulatory Activity of HT29-MTX Cells on Caco-2 Cell Growth and Differentiation
2.3. Quantitative RT-PCR Analyses
2.4. Real-Time Uptake of Oleic Acid by Caco-2, HT29-MTX Cells, and Co-Cultures
2.5. Transcriptional Regulations by Oleic Acid on Caco-2, HT29-MTX Cells and Co-Cultures
3. Discussion
4. Materials and Methods
4.1. Bio-Informatic Dataset Enrichment Analyses
4.2. Cell Culture
4.3. Real-Time Cell Analysis
4.4. Cell Survival and Size Analysis
4.5. Treatments
4.6. Histochemistry and Immunochemistry
4.7. Fluorescent Cell Lipid Storage Analysis
4.8. qRT-PCR Analysis of Gene Expression
5. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
Abbreviations
AB | Alcian Blue |
ABDH5 | α/β-hydrolase domain-containing protein 5 |
AMPK | Protein kinase AMP-activated |
APOA2 | Apoprotein 2 |
APOA3 | Apoprotein 3 |
ATOH1 (Hath1) | Helix-loop-helix (HLH) protein |
BMP4 | Bone Morphogenic Protein 4 |
BSA | Bovine serum albumin |
CIDEB and CIDEC | Cell-death activators CIDE-B and C |
DGAT2 | Diacylglycerol transferase 2 |
EGF | Epidermal growth Factor |
FABPs | Fatty acid binding proteins |
FASN | Fatty acid synthase |
FAT/CD36 | Fatty acid translocase |
Gfi1 | Growth factor independence 1 |
GPR120 | G-protein coupled receptor 120 |
HES1 | Hairy and enhancer of split-1 |
HPRT1 | Hypoxanthine phosphoribosyltransferase 1 |
Klf4 | Kruppel-like factor 4 |
LDLR | Low density lipid receptor |
LIPE | Lipid protein lipase |
MUC2 | Mucin 2 |
MUC5AC | Mucin 5AC |
OA | Oleic acid |
PAS | Periodic acid-Schiff reaction |
PLIN1, 3, 5 | Perilipins 1, 3 and 5 |
PPARs | Peroxisome proliferator activated receptors (alpha, beta/delta and gamma) |
qRT-PCR | Quantitative real-time polymerase chain reaction |
RTCA | Real-time Cell Analyzer |
SOX9 | SRY-Box 9 |
SPDEF | SAM Pointed Domain Containing ETS Transcription Factor |
TFF3 | Trefoil fator 3 |
TSPAN8 | Tetraspanin 8 |
XBP1 | X-box binding protein 1 |
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Gene Dataset Name | Gene Number | Dataset Analysis | GEO | Caco-2 Diff vs. Undiff | Caco-2 Diff vs. HT29-MTX | Caco Diff vs. Entero (ileal + Juj) | Colonocyte | Enterocyte |
---|---|---|---|---|---|---|---|---|
Common Gene Frequency % (Number) | ||||||||
Caco-2 differentiated vs. undifferentiated | 579 | Pellis et al., 2005 | GSE2047 | 13.6 (79) | 51.1 (296) | 1.9 (11) | 2.1 (12) | |
Caco-2 differentiated vs. HT29-MTX differentiated (3 weeks) | 1371 | −2 < FcLOG < 2 | GSE30292 | 5.8 (79) | 61.7 (846) | 0.9 (12) | 0.9 (13) | |
Caco-2 differentiated vs. (enterocyte ileal + jejunal) | 6038 | −2 < FcLOG < 2 | GSE30292 | 4.9 (296) | 14.0 (846) | 0.4 (22) | 0.4 (28) | |
Colonocyte | 103 | COPE (PubMed list) | 10.7 (11) | 11.7 (12) | 21.4 (22) | 48.5 (50) | ||
Enterocyte | 142 | COPE (PubMed list) | 8.5 (12) | 9.2 (13) | 19.7 (28) | 35.2 (50) |
Gene Symbol | Caco-2 | HT29-MTX | Caco-2/HT29-MTX | Fc vs. Theoric | Fc OA Caco-2/HT29-MTX |
---|---|---|---|---|---|
Intestinal phenotype matkers | |||||
ATOH1 (Hath1) | 1366 ± 30 | 17,481 ± 1590 | 212 ± 29 | −14 | 3.4 ± 0.2 |
GFi1 | 4.20 ± 0.10 | 56.0 ± 9.4 | 0.7 ± 0.1 | −14 | 3.9 ± 0.1 |
HES1 | 1260 ± 338 | 1697 ± 36 | 527 ± 29 | −2 | 1.6 ± 0.1 |
MUC2 | 17.4 ± 1.6 | 1.4 ± 0.2 | 2.0 ± 0.5 | −8 | 2.4 ± 1.7 |
MUC5 | 1540 ± 10 | 30,661 ± 1314 | 619 ± 27 | −7 | 4.2 ± 0.0 |
SOX9 | 228 ± 10 | 489 ± 83 | 158 ± 20 | −2 | −3.2 ± 0.0 |
TFF3 | 580774 ± 2 | 485,507 ± 15,791 | 101,000 ± 882 | −6 | 4.3 ± 0.2 |
Fatty acid uptake & transport | |||||
FAT/CD36 | 0.05 ± 0.03 | 1.32 ± 0.35 | 0.05 ± 0.00 | −3 | 1.0 ± 0.1 |
FABP1 | 263,971 ± 115,167 | 27 ± 2 | 50,176 ± 4078 | −5 | 4.9 ± 0.4 |
FABP3 | 191 ± 18 | 3.2 ± 0.5 | 98 ± 8 | −2 | 2.0 ± 0.5 |
FABP4 | 4.05 ± 0.62 | 0.69 ± 0.14 | 0.15 ± 0.03 | −25 | 26.7 ± 8.3 |
GRP120 | 2844 ± 91 | 9007 ± 473 | 312 ± 8 | −11 | 5.4 ± 0.2 |
Lipid droplet structure | |||||
ABDH5 (CGI-58) | 507 ± 158 | 165 ± 14 | 184 ± 28 | −3 | 3.7 ± 1.5 |
PLIN1 | nd | 0.03 ± 0.004 | 0.002 ± 0.001 | −1.6 ± 0.1 | |
PLIN2 (ADFP) | nd | nd | nd | ||
PLIN3 (TIP47) | 3199 ± 870 | 1670 ± 63 | 1200 ± 271 | −3 | 2.7 ± 0.1 |
PLIN5 (oxPAT) | 0.005 ± 0.001 | 0.031 ± 0.008 | 0.001 ± 0.000 | −10 | 1.9 ± 0.1 |
CIDEA | nd | nd | nd | ||
CIDEB | 16.1 ± 2.8 | 18.1 ± 2.0 | 3.1 ± 0.1 | −5 | 3.2 ± 0.2 |
CIDEC | 16,762 ± 2219 | 4837 ± 64 | 2217 ± 49 | −7 | 2.3 ± 0.2 |
UPR stress response | |||||
XBP1 | 19,033 ± 634 | 61,810 ± 2713 | 13,454 ± 6654 | −2 | −1.4 ± 0.0 |
Cancer marker | |||||
TSPAN8 | 124,836 ± 3825 | 706,666 ± 5070 | 19,088 ± 3500 | −10 | 4.7 ± 0.0 |
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Berger, E.; Nassra, M.; Atgié, C.; Plaisancié, P.; Géloën, A. Oleic Acid Uptake Reveals the Rescued Enterocyte Phenotype of Colon Cancer Caco-2 by HT29-MTX Cells in Co-Culture Mode. Int. J. Mol. Sci. 2017, 18, 1573. https://doi.org/10.3390/ijms18071573
Berger E, Nassra M, Atgié C, Plaisancié P, Géloën A. Oleic Acid Uptake Reveals the Rescued Enterocyte Phenotype of Colon Cancer Caco-2 by HT29-MTX Cells in Co-Culture Mode. International Journal of Molecular Sciences. 2017; 18(7):1573. https://doi.org/10.3390/ijms18071573
Chicago/Turabian StyleBerger, Emmanuelle, Merian Nassra, Claude Atgié, Pascale Plaisancié, and Alain Géloën. 2017. "Oleic Acid Uptake Reveals the Rescued Enterocyte Phenotype of Colon Cancer Caco-2 by HT29-MTX Cells in Co-Culture Mode" International Journal of Molecular Sciences 18, no. 7: 1573. https://doi.org/10.3390/ijms18071573