Related Experiment Video
Updated: Aug 10, 2026
High-Content Screening Assay for the Identification of Antibody-Dependent Cellular Cytotoxicity Modifying Compounds
Published on: August 18, 2023
Antiadhesive antibodies targeting E-cadherin sensitize multicellular tumor spheroids to chemotherapy in vitro
Shane K Green1, Giulio Francia, Ciro Isidoro
1Molecular and Cellular Biology Research, Sunnybrook and Women's College Health Sciences Center and Department of Medical Biophysics, University of Toronto, Toronto, ON, Canada.
Abstract:
Multicellular resistance, a subtype of therapeutic resistance manifested in cancer cells grown as three-dimensional multicellular masses, such as spheroids in vitro and solid tumors in vivo, occurs with respect to a variety of anticancer treatment strategies including chemotherapy, ionizing radiation, and even host-mediated antibody-dependent cellular cytotoxicity. Previous studies from our laboratory have shown that multicellular resistance to chemotherapy demonstrated by aggregates of EMT-6 murine mammary carcinoma cells can be overcome by using hyaluronidase to disrupt intercellular adhesive interactions and associated patterns of protein expression. In this proof of principle study, we explored the concept of antiadhesive chemosensitization in the context of human cancer cells by using a monoclonal antibody to disrupt E-cadherin-mediated cell-cell interactions in multicellular spheroids of HT29 human colorectal adenocarcinoma. In so doing, we found that disruption of E-cadherin-mediated adhesion sensitizes multicellular spheroids of HT29 in vitro to treatment with 5-fluorouracil, paclitaxel, vinblastine, and etoposide but not cisplatin. Furthermore, we have found that antibody-mediated blockage of E-cadherin function leads to decreased expression and activity of protein kinase C alpha and beta1, both of which have previously been implicated in chemoresistance exhibited by HT29 cells; however, we have found that the chemosensitization effects of the anti-E-cadherin antibody are independent of its influence on protein kinase C beta1.
Insights
Disrupting E-cadherin interactions with an antibody sensitizes human colorectal cancer spheroids to several chemotherapy drugs. This anti-adhesive strategy overcomes multicellular resistance, offering new therapeutic approaches.
Area of Science:
- Oncology
- Cancer Biology
- Pharmacology
Background:
- Multicellular resistance is a significant challenge in cancer therapy, affecting responses to chemotherapy, radiation, and immunotherapy.
- Previous research demonstrated hyaluronidase can overcome multicellular resistance in murine models by disrupting cell adhesion.
- E-cadherin-mediated cell-cell interactions are crucial for the structural integrity and therapeutic resistance of multicellular cancer aggregates.
Purpose of the Study:
- To investigate the efficacy of antiadhesive chemosensitization in human colorectal cancer cells.
- To determine if blocking E-cadherin-mediated adhesion sensitizes HT29 human colorectal adenocarcinoma spheroids to chemotherapy.
- To explore the role of protein kinase C (PKC) signaling in E-cadherin-mediated chemoresistance.
Main Methods:
- Utilized HT29 human colorectal adenocarcinoma cells cultured as multicellular spheroids.
- Employed a monoclonal antibody to disrupt E-cadherin-mediated cell-cell interactions.
- Assessed spheroid chemosensitivity to various chemotherapeutic agents (5-fluorouracil, paclitaxel, vinblastine, etoposide, cisplatin).
- Quantified changes in protein kinase C alpha and beta1 expression and activity.
Main Results:
- Disruption of E-cadherin adhesion sensitized HT29 spheroids to 5-fluorouracil, paclitaxel, vinblastine, and etoposide.
- Cisplatin efficacy was not enhanced by E-cadherin blockade.
- Antibody-mediated E-cadherin blockage reduced the expression and activity of protein kinase C alpha and beta1.
- The observed chemosensitization was independent of the antibody's effect on protein kinase C beta1.
Conclusions:
- Antiadhesive strategies targeting E-cadherin can overcome multicellular resistance in human colorectal cancer spheroids.
- This approach enhances sensitivity to specific chemotherapeutic agents, suggesting a potential new therapeutic avenue.
- While E-cadherin blockade affects PKC signaling, the chemosensitization mechanism is not solely dependent on PKC beta1 activity.
More Related Videos
13:19Quantifying Antibody-Dependent Cellular Cytotoxicity in a Tumor Spheroid Model: Application for Drug Discovery
Published on: April 26, 2024
05:21Assessing Antibody-dependent, Cell-mediated Cytotoxicity in Cancer Cells using Antibody-Dependent Cell-Mediated Cytotoxicity Reporter Bioassay
Published on: September 13, 2024
