Related Experiment Video
Updated: Aug 9, 2026
In vitro Organoid Culture of Primary Mouse Colon Tumors
Published on: May 17, 2013
Multidrug-resistant phenotype influences the differentiation of a human colon carcinoma cell line
1UPRESA-CNRS 6032, UFR Pharmacie, 27 Bd J. Moulin, Marseille, 13005, France.
Abstract:
The human colon carcinoma cell line HT29-D4, which constitutively expresses a very low level of the MDR1 gene product, was made multidrug resistant by transfection with a human MDR1 cDNA from the pHaMDR1/A expression vector and selection by colchicine. Resistant clones were 3- to 15-fold resistant to colchicine and were cross-resistant to doxorubicin (3- to 4-fold). MDR1 gene expression was associated with the expression of functional P-glycoprotein (gp-170); the function was reversed by verapamil and cyclosporin A. HT29-D4 cells are able to differentiate in vitro by replacement of glucose by galactose in the culture medium and also to release the carcinoembryonic antigen (CEA). Under these culture conditions, MDR1 mRNA and gp-170 were always expressed and the protein remained functional. Upon galactose treatment, resistant clones were less differentiated since they showed a heterogeneous monolayer organization accompanied by heterogeneous staining of cell-surface CEA and a high decrease (60-90%) of CEA release.
Insights
Multidrug resistance in colon cancer cells (HT29-D4) was achieved by introducing the MDR1 gene. This enhanced resistance to chemotherapy drugs like colchicine and doxorubicin, even during cell differentiation.
Area of Science:
- Molecular biology
- Cancer research
- Cell biology
Background:
- The HT29-D4 human colon carcinoma cell line exhibits low basal expression of the multidrug resistance (MDR1) gene.
- Multidrug resistance is a significant challenge in cancer chemotherapy, limiting treatment efficacy.
Purpose of the Study:
- To establish a multidrug-resistant colon cancer cell line (HT29-D4) by overexpressing the MDR1 gene.
- To investigate the functional expression of P-glycoprotein (gp-170) in resistant cells.
- To assess the impact of in vitro differentiation on multidrug resistance and P-glycoprotein function.
Main Methods:
- Transfection of HT29-D4 cells with human MDR1 cDNA using the pHaMDR1/A vector.
- Selection of resistant clones using colchicine.
- Assessment of drug resistance (colchicine, doxorubicin) and P-glycoprotein expression (gp-170).
- In vitro differentiation induced by replacing glucose with galactose in culture medium.
- Measurement of carcinoembryonic antigen (CEA) release as a differentiation marker.
Main Results:
- Resistant clones showed 3- to 15-fold resistance to colchicine and 3- to 4-fold cross-resistance to doxorubicin.
- MDR1 gene expression correlated with functional P-glycoprotein (gp-170) expression, reversible by verapamil and cyclosporin A.
- During galactose-induced differentiation, MDR1 mRNA and functional gp-170 remained expressed.
- Resistant cells exhibited reduced differentiation, characterized by heterogeneous cell organization and decreased CEA release (60-90%) upon galactose treatment.
Conclusions:
- Successful generation of a multidrug-resistant HT29-D4 colon cancer cell line via MDR1 gene transfection.
- P-glycoprotein remains functional during in vitro differentiation, suggesting MDR1 expression is independent of this differentiation pathway.
- Galactose-induced differentiation in resistant cells leads to impaired differentiation markers and reduced CEA release.
More Related Videos
08:46Implementation of In Vitro Drug Resistance Assays: Maximizing the Potential for Uncovering Clinically Relevant Resistance Mechanisms
Published on: December 9, 2015
07:20Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium
Published on: April 9, 2019
Related Concept Videos
02:56Treatment Resistant Cancers
01:11Pharmacogenetics of Drug Targets: β₂-Adrenergic Receptors, Apo E, Thymidylate Synthase