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Updated: Dec 6, 2025
A Robust Discovery Platform for the Identification of Novel Mediators of Melanoma Metastasis
Published on: March 8, 2022
Analysis of candidate genes expected to be essential for melanoma surviving
Irina A Krivosheeva1, Alexandra Yu Filatova1, Sergei A Moshkovskii2
1Laboratory of Functional Genomics, Research Centre of Medical Genetics, Erevanskaya Street, 10 building 2, Floor 44, Moscow, 115304 Russia.
Introduction:
Cancers may be treated by selective targeting of the genes vital for their survival. A number of attempts have led to discovery of several genes essential for surviving of tumor cells of different types. In this work, we tried to analyze genes that were previously predicted to be essential for melanoma surviving. Here we present the results of transient siRNA-mediated knockdown of the four of such genes, namely, UNC45A, STK11IP, RHPN2 and ZNFX1, in melanoma cell line A375, then assayed the cells for their viability, proliferation and ability to migrate in vitro. In our study, the knockdown of the genes predicted as essential for melanoma survival does not lead to statistically significant changes in cell viability. On the other hand, for each of the studied genes, mobility assays showed that the knockdown of each of the target genes accelerates the speed of cells migrating. Possible explanation for such counterintuitive results may include insufficiency of the predicting computational models or the necessity of a multiplex knockdown of the genes.
Aims:
To examine the hypothesis of essentiality of hypomutated genes for melanoma surviving we have performed knockdown of several genes in melanoma cell line and analyzed cell viability and their ability to migrate.
Methods:
Knockdown was performed by siRNAs transfected by Metafectene PRO. The levels of mRNAs before and after knockdown were evaluated by RT-qPCR analysis. Cell viability and proliferation were assessed by MTT assay. Cell migration was assessed by wound healing assay.
Results:
The knockdown of the genes predicted as essential for melanoma survival does not lead to statistically significant changes in cell viability. On the other hand, for each of the studied genes, mobility assays showed that the knockdown of each of the target genes accelerates the speed of cells migrating.
Conclusion:
Our results do not confirm initial hypothesis that the genes predicted essential for melanoma survival as a matter of fact support the survival of melanoma cells.
Insights
Targeting genes essential for melanoma survival did not impact cell viability. However, gene knockdown accelerated melanoma cell migration, challenging current predictive models for cancer gene essentiality.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Cancer treatment strategies focus on targeting genes crucial for tumor cell survival.
- Previous computational models predicted several genes essential for melanoma survival.
Purpose of the Study:
- To investigate the essentiality of predicted genes for melanoma survival.
- To analyze the effect of gene knockdown on melanoma cell viability and migration.
Main Methods:
- Transient siRNA-mediated knockdown of UNC45A, STK11IP, RHPN2, and ZNFX1 in A375 melanoma cells.
- Assessment of cell viability, proliferation (MTT assay), and migration (wound healing assay).
- mRNA level evaluation using RT-qPCR.
Main Results:
- Knockdown of predicted essential genes did not significantly alter melanoma cell viability or proliferation.
- Migration assays revealed accelerated cell movement upon knockdown of each target gene.
Conclusions:
- The study challenges the hypothesis that the investigated genes are essential for melanoma cell survival.
- Observed increase in cell migration suggests potential limitations in predictive models or the need for multiplex gene targeting.
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