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Updated: Jun 20, 2026
Heterotypic Three-dimensional In Vitro Modeling of Stromal-Epithelial Interactions During Ovarian Cancer Initiation and Progression
Published on: August 28, 2012
Blockage of transdifferentiation from fibroblast to myofibroblast in experimental ovarian cancer models
1Division of Molecular and Gene Therapies, Griffith Institute for Health and Medical Research, School of Medical Science, Griffith University, Gold Coast campus, Southport, Qld 4222, Australia.
Background:
Tumour stromal myofibroblasts can promote tumour invasion. As these cells are genetically more stable than cancer cells, there has been enormous interest in developing targeted molecular therapies against them. Chloride intracellular channel 4 (CLIC4) and reactive oxygen species (ROS) have been linked with promoting stromal cell transdifferentiation in various cancers, but little is known of their roles in ovarian cancer. In this study, we examined the functional roles that both CLIC4 and ROS play in the process of ovarian cancer cell-stimulated or TGF-beta1 induced fibroblast-to-myofibroblast transdifferentiation. We also examine whether it is possible to reverse such a process, with the aim of developing novel therapies against ovarian cancer by targeting activated transdifferentiated myofibroblasts.
Results:
We demonstrate that TGF-beta1 induced or CM(SKOV3) activate transdifferentiated myofibroblasts (fibroblasts). These fibroblasts mimic "reactive" stromal myofibroblasts and demonstrate significant up-regulation of CLIC4 expression and increased level of ROS production. Blocking the production of ROS with an antioxidant consequently reduces the expression of CLIC4, and is accompanied by disappearance of alpha-smooth-muscle actin (alpha-SMA), a myofibroblast marker, suggesting ROS acts as a signalling molecule that promotes and enhances CLIC4 activities in the myofibroblast transdifferentiaton process. Down-regulation of CLIC4 with a generic agent or specific siRNA both significantly reduces the expression of factors related to the phenotypes and functions of myofibroblasts, such as alpha-SMA, hepatocyte growth factor (HGF) and vascular endothelial growth factor (VEGF), thus reversing the myofibroblast phenotype back to fibroblasts. These results convincingly show that ROS and CLIC4 are responsible for TGF-beta1 induced fibroblast-to-myofibroblast transdifferentiaton and down-regulation of both is sufficient to block transdifferentiated myofibroblasts.
Conclusion:
Molecular targeting of ROS and CLIC4 has the potential to develop novel therapies for ovarian cancer.
Insights
Targeting reactive oxygen species (ROS) and Chloride Intracellular Channel 4 (CLIC4) can reverse ovarian cancer fibroblast activation. This finding offers potential for novel ovarian cancer therapies.
Area of Science:
- Oncology
- Cell Biology
- Molecular Medicine
Background:
- Tumor-associated myofibroblasts promote cancer invasion.
- Chloride Intracellular Channel 4 (CLIC4) and reactive oxygen species (ROS) are implicated in stromal cell transdifferentiation.
- The roles of CLIC4 and ROS in ovarian cancer remain largely unknown.
Purpose of the Study:
- To investigate the functional roles of CLIC4 and ROS in ovarian cancer fibroblast-to-myofibroblast transdifferentiation.
- To explore the potential for reversing myofibroblast activation for novel ovarian cancer therapies.
Main Methods:
- Induction of fibroblast-to-myofibroblast transdifferentiation using TGF-beta1 or cancer cell-conditioned media.
- Assessment of CLIC4 expression and ROS production.
- Inhibition of ROS using antioxidants.
- Down-regulation of CLIC4 using generic agents and siRNA.
- Analysis of myofibroblast markers (e.g., alpha-SMA) and functional factors (e.g., HGF, VEGF).
Main Results:
- TGF-beta1 or cancer cell-conditioned media induced myofibroblast activation with increased CLIC4 and ROS.
- ROS inhibition reduced CLIC4 expression and alpha-SMA, indicating ROS signaling in transdifferentiation.
- CLIC4 down-regulation reversed myofibroblast phenotype by reducing alpha-SMA, HGF, and VEGF.
- Targeting ROS and CLIC4 effectively blocked myofibroblast activation.
Conclusions:
- ROS and CLIC4 are key drivers of fibroblast-to-myofibroblast transdifferentiation in ovarian cancer.
- Targeting ROS and CLIC4 presents a promising therapeutic strategy for ovarian cancer.
