Biopolym. Cell. 2020; 36(2):99-109.
http://dx.doi.org/10.7124/bc.000A24
Structure and Function of Biopolymers
The biological properties of HEK293T cell line transfected with mCD150 and nCD150 isoforms of CD150/SLAMF1 receptor
1Shlapatska L. M., 1Gordiienko I. M., 1Kovalevska L. M., 1Sidorenko S. P.
  1. R. E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology, NAS of Ukraine
    45, Vasilkivska Str., Kyiv, Ukraine, 01022

Abstract

Aim. To elucidate the role of CD150/SLAMF1 receptor isoforms mCD150 and nCD150 in regulation of HEK293T cells proliferation activity and clonogenicity. Methods. Cell culture, transfection, qPCR, flow cytometry, cells proliferationandcell cycle analysis, CFU assay and statistical analysis were used. Results. As a result of transfection of HEK293T cell line with mCD150 or nCD150 cDNA, two cell clones were obtained with stable and exclusively cytoplasmic expression of CD150 isoforms – HEK293T-pBABE-mCD150 and HEK293T-pBABE-nCD150. HEK293T cells transfected with mCD150 and nCD150 isoforms are characterized by higher proliferation supported by an elevated level of the IPO38 antigen expression compared to HEK293T and HEK293T-pBABE-puro. The analysis of cell cycle revealed a significantly higher percentage of HEK293T-pBABE-mCD150 and HEK293T-pBABE-nCD150 cells in S phase and lower in G0/G1 in contrast to HEK293T and HEK293T-pBABE-puro. Compared to the control cell line, a significantly elevated percentage of HEK293Tstablytransfected with the nCD150, but not with the mCD150 isoform, were observed in G2/M phase of cell cycle. Both HEK293T-pBABE-mCD150 and HEK293T-pBABE-nCD150 cells were able to form colonies at a low cell density, contrary to HEK293T and HEK293-pBABE-puro. Conclusion. Both mCD150 and nCD150 isoforms possess cell-growth promoting properties stimulating cell proliferation, cell cycle progression and clonogenic potential with more profound effect of the nCD150 isoform.
Keywords: CD150/SLAMF1 receptor, mCD150 isoform, nCD150 isoform, HEK293T cell line, cell proliferation, clonogenicity
Full text: (PDF, in English)

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