[Screening of drugs that selectively inhibit uveal melanoma cells with SF3B1 mutations]
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Updated: Oct 7, 2025
Implantation and Evaluation of Melanoma in the Murine Choroid via Optical Coherence Tomography
Published on: December 2, 2022
1Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Objective:
To screen compounds that can selectively inhibit uveal melanoma cells with splicing factor 3B subunit 1 (SF3B1) mutations in comparison with isogenic SF3B1 wild-type counterparts in a cell model of SF3B1 mutant allele knockout.
Methods:
Principal component analysis was used to analyze transcriptome alternative splicing in TCGA cohorts of uveal melanoma with wild-type SF3B1 and SF3B1 mutations, and abnormal alternative splicing events derived from SF3B1 mutations were identified. The SF3B1 mutant allele in Mel202 cells was knocked out using CRISPR-Cas9 technology, and Sanger sequencing was used to verify the edited sequence. MTT and colony formation assays were used to assess the proliferation of Mel202 and Mut-KO cells. RT-PCR agarose electrophoresis combined with Sanger sequencing was used to determine alternative splicing events in Mel202 and Mut-KO cells. MTT assay was performed to screen the compounds that showed selective inhibitory effect against Mel202 cells with SF3B1 mutation.
Results:
Specific knockout of SF3B1 mutant allele in Mel202 cells obviously promoted the cell proliferation and caused changes in alternative splicing of ZDHHC16 and DYNLL1 transcripts. The screening data showed that 13 compounds had selective inhibitory activity against Mel202 cells with SF3B1 mutation (Fold change≥2), and among them, tetrandrine and lapatinib showed good dose-effect curves.
Conclusion:
This study provides a cell screening model for identification of potential individualized treatment drugs for patients with uveal melanoma with SF3B1 mutation.
Insights
Researchers screened compounds to target uveal melanoma cells with SF3B1 mutations. Tetrandrine and lapatinib showed selective inhibitory activity, offering potential for individualized treatments in SF3B1-mutated uveal melanoma.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Uveal melanoma (UM) is the most common primary intraocular cancer in adults.
- Mutations in splicing factor 3B subunit 1 (SF3B1) are common in UM and impact alternative splicing.
- Targeting SF3B1 mutations offers a potential therapeutic strategy for UM.
Purpose of the Study:
- To develop and utilize a cell model for screening compounds selectively inhibiting UM cells with SF3B1 mutations.
- To identify novel therapeutic agents for UM patients harboring SF3B1 mutations.
Main Methods:
- Transcriptome alternative splicing analysis using TCGA UM cohorts.
- CRISPR-Cas9 technology to create an SF3B1 mutant allele knockout cell model (Mel202 vs. Mut-KO).
- MTT and colony formation assays to assess cell proliferation and compound efficacy.
- RT-PCR and Sanger sequencing to analyze alternative splicing events.
Main Results:
- SF3B1 mutation knockout in Mel202 cells promoted proliferation and altered alternative splicing of ZDHHC16 and DYNLL1.
- Thirteen compounds demonstrated selective inhibitory activity against SF3B1-mutated UM cells.
- Tetrandrine and lapatinib exhibited significant dose-dependent inhibitory effects.
Conclusions:
- A novel cell screening model was established for identifying targeted therapies for SF3B1-mutated UM.
- Tetrandrine and lapatinib are promising candidates for further investigation as individualized treatments for UM with SF3B1 mutations.
