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Reduced Expression of PEDF and ALDH1A1 during Spheroid Transition of Lung Cancer Cells: An In Vitro Study

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Abstract

“Cancer stem cells” (CSCs) can initiate tumorigenesis and metastasis and show resistance against chemotherapy owing to the expression of prominent stem cell markers. CSCs are a subpopulation of highly heterogenic cancer spheroid cells. Pigment epithelium-derived factor (PEDF) is a neurotrophic, anti-tumorigenic, and anti-metastatic protein and its gene expression levels in A549 spheroids is still unknown. We aimed to compare clonogenicity and mRNA levels of PEDF, Oct4, and ALDH1A1 between A549 and spheroid cells. Spheroid and colony formation assays were performed with spheroid and A549 cells. We performed quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for gene expression analysis. The clonogenic ratios for A549 and spheroids were ~60% and ~1% respectively. During spheroid formation, Oct4 mRNA level did not change but PEDF and ALDH1A1 levels decreased significantly. CSCs are characterized by elevated stem cell markers but spheroid cells consist of heterogeneous population including CSCs. In spheroid population, no increase in stem cell markers was observed. The reduced PEDF levels during spheroid transition can be a suppression mechanism of spheroid cells.

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ACKNOWLEDGMENTS

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Funding

This research was funded by the Scientific Research Project Funds of Hatay Mustafa Kemal University (Project no. 18.M. 102).

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Authors and Affiliations

  1. Department of Medical Biology, Faculty of Medicine, Hatay Mustafa Kemal University, Hatay, Turkey

    M. Y. Terzi, G. Gülbol-Duran & M. Urhan-Küçük

  2. Department of Molecular Biochemistry and Genetics, Graduate School of Health Sciences, Hatay Mustafa Kemal University, Hatay, Turkey

    M. Y. Terzi, G. Gülbol-Duran & M. Urhan-Küçük

  3. Department of Medical Services and Techniques, Vocational School of Health Services, Hatay Mustafa Kemal University, Hatay, Turkey

    H. M. Okuyan

  4. Department of Physiotherapy and Rehabilitation, Faculty of Health Sciences, Sakarya University of Applied Sciences, Sakarya, Turkey

    H. M. Okuyan

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  1. M. Y. Terzi
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Correspondence to M. Y. Terzi.

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The authors declare that they have no conflicts of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

AUTHOR CONTRIBUTION

MYT designed the study. MYT and HMO performed the experiments. M.Y.T., H.M.O., G.G.D., and M.U.K. analyzed the data statistically and M.Y.T. prepared all figures. M.Y.T. prepared tables. All authors wrote/drafted/edited/revised the manuscript and interpreted the results. All authors read, edited, and gave an approval for the final version of the present manuscript. All authors are aware of the order of authorship and that no further changes in authorship will be performed after submission.

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Terzi, M.Y., Okuyan, H.M., Gülbol-Duran, G. et al. Reduced Expression of PEDF and ALDH1A1 during Spheroid Transition of Lung Cancer Cells: An In Vitro Study. Cytol. Genet. 56, 172–178 (2022). https://doi.org/10.3103/S0095452722020104

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