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Updated: Aug 15, 2025
Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
Published on: September 20, 2016
FAM111B dysregulation promotes malignancy in fibrosarcoma and POIKTMP and a low-cost method for its mutation
Cenza Rhoda1, Falone Sunda1, Elvis Kidzeru1
1Hair and Skin Research Laboratory, Division of Dermatology, Department of Medicine, Faculty of Health Sciences, University of Cape Town, South Africa.
Introduction:
Mutations in the uncharacterised human FAM111B gene are associated with POIKTMP, a rare multi-organ fibrosing disease. Recent studies also reported the overexpression of FAM111B in specific cancers. Moreover, FAM111B mutation screening may prove expensive in under-resourced facilities. Therefore, this study investigated its cellular function and dysfunction and described an inexpensive mutation screening method.
Materials And Methods:
FAM111B expression was assessed in silico and validated in vitro in cell lines and primary skin fibroblasts from a South African POIKTMP-patient with the heterozygous FAM111B gene mutation: NM_198947.4: c.1861T>G (p. Tyr621Asp or Y621D) by qPCR and western blot. The cellular function of FAM111B was studied in HT1080 using various cell-based functional assays, and the Y621D mutation was genotyped by PCR-RFLP.
Results:
Expression studies showed upregulated FAM111B mRNA and protein in the cancer cells. High FAM111B expression with robust nuclear localization occurred in HT1080. Additionally, expression data and cell-based assays indicated that FAM111B led to the upregulation of cell migration, decreased cell apoptosis, and modulatory effects on cell proliferation. Y621D mutation showed similar effects on cell migration but minimal impact on cell apoptosis. FAM111B mRNA and protein expression were markedly downregulated (p ≤ 0.05) in the POIKTMP-patient's fibroblasts. The PCR-RFLP method successfully genotyped Y621D gene mutation.
Discussion:
FAM111B is a cancer-associated nuclear protein: Its modulation by mutations or overexpression may contribute to the malignancy of cancers and POIKTMP/fibrosis and poor clinical outcomes and represents a viable prognostic marker or therapeutic target. Furthermore, the PCR-RFLP method could prove a valuable tool for FAM111B mutation validation or screening in resource-constrained laboratories.
Insights
FAM111B gene mutations are linked to POIKTMP and cancer. This study reveals FAM111B
Area of Science:
- Genetics
- Molecular Biology
- Oncology
Background:
- Mutations in the FAM111B gene are associated with POIKTMP, a rare fibrosing disease.
- FAM111B overexpression is observed in certain cancers.
- Current mutation screening methods for FAM111B can be costly.
Purpose of the Study:
- To investigate the cellular function and dysfunction of FAM111B.
- To describe an inexpensive method for FAM111B mutation screening.
Main Methods:
- FAM111B expression was analyzed in silico and validated in vitro using qPCR and western blot.
- Cellular functions were assessed using cell-based assays in HT1080 cells.
- The Y621D mutation was genotyped using PCR-RFLP.
Main Results:
- FAM111B expression was upregulated in cancer cells, promoting migration and affecting proliferation and apoptosis.
- The Y621D mutation impacted cell migration similarly to wild-type FAM111B but had minimal effect on apoptosis.
- FAM111B expression was downregulated in POIKTMP patient fibroblasts, and the PCR-RFLP method successfully identified the mutation.
Conclusions:
- FAM111B is a cancer-associated nuclear protein; its modulation may drive cancer and fibrosis, serving as a prognostic marker or therapeutic target.
- The developed PCR-RFLP method offers a cost-effective tool for FAM111B mutation screening in resource-limited settings.
