Establishment and characterization of stable cell-based model expressing Gfp/ SQSTM1/p62 for autophagy modulator screening

  • wipawan sirigulpanit faculty of pharmaceutical sciences, Burapha University
  • Pathompong Bowornruangrit
Keywords: Autophagy, Autophagy modulator, Stable cell-based model expressing Gfp/SQSTM1/p62

Abstract

Autophagy plays a key role in a number of diseases, including cancer. This study aimed to generate a stable BHK-21 cell line containing a green fluorescent protein gene (Gfp) and sequestosome 1 gene (SQSTM1/p62) for a cell-based model to screen autophagy modulator. Stable BHK-21 (Gfp/SQSTM1/p62) cell line was established in this study then examined for effectiveness in an autophagy modulator screening assay using rapamycin and chloroquine known autophagy modulators. The SQSTM1/p62 protein turnover has been measured as autophagy activity. The results showed that SQSTM1/p62 protein is active in normal growing conditions at basal level. An induction of autophagy, however, increase the degradation of SQSTM1/p62 protein causes SQSTM1/p62 protein reduction in cells while a decrease or block of this pathway causes SQSTM1/p62 protein accumulation which increases SQSTM1/p62 protein level in cells. This study indicated that stable BHK-21 (Gfp/SQSTM1/p62) cell line is available and efficient for use in an autophagy activity analysis.

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Published
2021-04-30
How to Cite
1.
sirigulpanit wipawan, Bowornruangrit P. Establishment and characterization of stable cell-based model expressing Gfp/ SQSTM1/p62 for autophagy modulator screening . Naresaun Phayao J [Internet]. 2021Apr.30 [cited 2021Sep.23];14(1):33-6. Available from: https://li01.tci-thaijo.org/index.php/journalup/article/view/247080
Section
Research articles