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Fhit loss-associated initiation and progression of neoplasia in vitro
Jenna R Karras1, Morgan S Schrock1, Bahadir Batar1
1Department of Cancer Biology and Genetics, Ohio State University Wexner Medical Center, Columbus, Ohio, USA.
Abstract:
The FHIT gene, encompassing an active common fragile site, FRA3B, is frequently silenced in preneoplasia and cancer, through gene rearrangement or methylation of regulatory sequences. Silencing of Fhit protein expression causes thymidine kinase 1 downregulation, resulting in dNTP imbalance, and spontaneous replication stress that leads to chromosomal aberrations, allele copy number variations, insertions/deletions, and single-base substitutions. Thus, Fhit, which is reduced in expression in the majority of human cancers, is a genome "caretaker" whose loss initiates genome instability in preneoplastic lesions. To follow the early genetic alterations and functional changes induced by Fhit loss that may recapitulate the neoplastic process in vitro, we established epithelial cell lines from kidney tissues of Fhit-/- and +/+ mouse pups early after weaning, and subjected cell cultures to nutritional and carcinogen stress, which +/+ cells did not survive. Through transcriptome profiling and protein expression analysis, we observed changes in the Trp53/p21 and survivin apoptotic pathways in -/- cells, and in expression of proteins involved in epithelial-mesenchymal transition. Some Fhit-deficient cell lines showed anchorage-independent colony formation and increased invasive capacity in vitro. Furthermore, cells of stressed Fhit-/- cell lines formed s.c. and metastatic tumors in nude mice. Collectively, we show that Fhit loss and subsequent thymidine kinase 1 inactivation, combined with selective pressures, leads to neoplasia-associated alterations in genes and gene expression patterns in vitro and in vivo.
Insights
Loss of the FHIT gene, a genome caretaker, causes instability and initiates cancer. Fhit loss leads to DNA damage and mutations, promoting tumor development in mice.
Area of Science:
- Genetics
- Cancer Biology
- Molecular Oncology
Background:
- The FHIT gene, located at a common fragile site (FRA3B), is frequently silenced in preneoplasia and cancer.
- Loss of Fhit protein expression results in thymidine kinase 1 downregulation, leading to dNTP imbalance and replication stress.
- Fhit acts as a genome caretaker, and its loss initiates genome instability, contributing to early neoplastic lesions.
Purpose of the Study:
- To investigate early genetic and functional changes induced by Fhit loss in vitro.
- To model the neoplastic process following Fhit deficiency using mouse kidney epithelial cell lines.
- To understand how Fhit loss, combined with stress, contributes to tumor initiation and progression.
Main Methods:
- Established epithelial cell lines from Fhit-/- and +/+ mouse pups.
- Subjected cell cultures to nutritional and carcinogen stress.
- Utilized transcriptome profiling and protein expression analysis.
- Assessed anchorage-independent colony formation and invasive capacity in vitro.
- Evaluated tumor formation in nude mice models.
Main Results:
- Fhit loss induced changes in Trp53/p21 and survivin apoptotic pathways.
- Observed alterations in proteins involved in epithelial-mesenchymal transition.
- Fhit-deficient cells exhibited anchorage-independent growth and increased invasion in vitro.
- Stressed Fhit-/- cells formed tumors and metastatic lesions in vivo.
Conclusions:
- Fhit loss, coupled with thymidine kinase 1 inactivation and selective pressures, drives neoplasia-associated alterations.
- Fhit deficiency promotes genome instability and contributes to the initiation and progression of cancer.
- The study demonstrates a causal link between Fhit loss and the development of cancer phenotypes in vitro and in vivo.
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