The Change of Podocyte β-Catenin by Puromycin Aminonucleoside

Choi, Ji-Young; Ahn, Eun-Mi; Park, Hye-Young; Shin, Jae-Il; Ha, Tae-Sun

doi:2011.15.2.138

J Korean Soc Pediatr Nephrol > Volume 15(2); 2011 > Article

J Korean Soc Pediatr Nephrol 2011;15(2): 138-145. doi: https://doi.org/10.3339/jkspn.2011.15.2.138

Puromycin aminonucleoside 투여에 따른 사구체 족세포 β-catenin의 변화

최지영, 안은미, 박혜영, 신재일, 하태선

¹충북대학교 의과대학 소아과학교실
²충북대학교 의과대학 소아과학교실
³충북대학교 의과대학 소아과학교실
⁴연세대학교 의과대학 소아과학교실
⁵충북대학교 의과대학 소아과학교실

The Change of Podocyte β-Catenin by Puromycin Aminonucleoside

Ji-Young Choi, Eun-Mi Ahn, Hye-Young Park, Jae-Il Shin, Tae-Sun Ha

¹Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
²Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
³Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease
⁴Department of Pediatrics, Yonsei University College of Medicine, Severance Children's Hospital
⁵Department of Pediatrics, College of Medicine, Chungbuk National University Cheongju, The Institute of Kidney Disease

Received: September 1, 2011; Accepted: October 4, 2011.

ABSTRACT

Purpose : To test whether the expression of ${beta}$-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro.
Methods : We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of ${beta}$-catenin by confocal microscope and measured the change of ${beta}$-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR).
Results :We found that ${beta}$-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN ($50{mu}g/mL$) decreased ${beta}$-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations ($50{mu}g/mL$) of PAN also decreased ${beta}$-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05).
Conclusion : Exposure of podocytes to PAN in vitro relocates ${beta}$-catenin internally and reduces ${beta}$-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.

Key words: ${\beta}$-Catenin | Puromycin aminonucleoside (PAN)-induced nephropathy | Glomerular epithelial cells (GEpC) | Podocyte