Abstract
We have recently found that primary rat embryonic fibroblasts (REFs) could be immortalized by overexpression of the human mitochondrial ribosomal protein MRPS18-2 (S18-2). A derived cell line, designated 18IM, expressed the embryonic stem cell markers SSEA-1 and Sox2. Upon inoculation into severe combined immunodeficiency mice, 18IM cells differentiated to express pan-keratin. They were not tumorigenic. Here we report the gene profiling of 18IM, compared with REF cells. Pathways involved in oxidative phosphorylation, ubiquinone (Coenzyme Q 10) biosynthesis, fatty acid elongation in mitochondria, PI3K/AKT signaling, a characteristic of rapidly proliferating cells, were upregulated in 18IM. Genes involved in the transcription/translation machinery and redox reactions, like elongation factors, ATP synthases, NADH dehydrogenases, mitogen activated kinases were upregulated as well. 18IM cells produced more pyruvate, indicating enhanced ATP synthesis. The expression of Oct4, Sox2, and Nanog that can contribute to the experimental induction of pluripotency in primary fibroblasts was also elevated, in contrast to Klf4 and C-myc that were downregulated. Subsequently, three new immortalized cell lines were produced by S18-2 overexpression in order to check the representativeness of 18IM. All of them showed anchorage-independent growth pattern. Two of three clones lost vimentin and smooth muscle actin, and expressed Sox2 and Oct4. We suggest that S18-2 is involved in the developmental regulation.
Highlights
OCT4 and SOX2 could induce pluripotency in human fibroblasts by themselves, in conjunction with valproic acid, a histone deacetylase inhibitor.[7]
We have accidentally discovered that overexpression of the human mitochondrial ribosomal protein MRPS18-2 (S18-2, NP_054765) immortalized primary rat embryonic fibroblasts (REFs).[10]
The MRPS18-2 (MRSPS18B, S18-2) protein is encoded by a gene on human chromosome 6p21. 3, adjacent to the MHC class II gene cluster
Summary
OCT4 and SOX2 could induce pluripotency in human fibroblasts by themselves, in conjunction with valproic acid, a histone deacetylase inhibitor.[7]. We have accidentally discovered that overexpression of the human mitochondrial ribosomal protein MRPS18-2 (S18-2, NP_054765) immortalized primary rat embryonic fibroblasts (REFs).[10] The derived cell line, designated 18IM, lost its sensitivity to contact inhibition and acquired the ability for anchorage-independent growth in soft agar with a cloning efficiency of 94%. It expressed the embryonic stem cell markers SSEA-1 and Sox[2] that were not detected in the original REFs, nor in C-MYC- and HA-RAS-transformed. We report the gene expression pattern of 18IM cells and three new S18-2-immortalized cell lines, in comparison with control REFs
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