在分子水平上研究BOLL表現與精母細胞分化的關聯性

林宗彥^1.2、鄭詠璇²、陳幸儀²、侯琇瓊²、林世杰³、林永明²、鄭裕生²

¹國立成功大學附設醫院斗六分院泌尿部、²國立成功大學附設醫院泌尿部、³國立成功大學基礎醫學研究所

The association between BOLL expression and the spermatocyte cell differentiation at the molecular level  

Tsung-Yen Lin^1,2, Yung-Hsuan Cheng², Hsing-Yi Chen², Hsiu-Chiung Hou², Shih-ChiehLin², Yung-Ming Lin³, Yu-Sheng Cheng³

¹Division of Urology, Department of Surgery, National Cheng Kung University Hospital Dou-Liou Branch, Yunlin, Taiwan, ²Department of Urology, Medical College and Hospital, National Cheng-Kung University, Tainan, Taiwan, and ³Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan

Abstract

Purpose:

To investigate whether BOLL would improve the spermatocyte cell differentiation at the molecular level

Materials and Methods:

Mouse spermatocyte cell line (GC2-spd) and human embryonic kidney cells 293 (Hek293T) were used in this study. BOLL expression was assessed after adding Aza-deoxycytidine in GC2 cell and transfection of whole BOLL sequence in Hek293T cell. In both cell lines, spermatid markers including PRM2, GAPDHS, TNP1, TNP2, CATSPER1, and CATSPER3 were evaluated. In addition, spermatogonia makers, including ID4 and PIWI4, were assessed in Hek293T cells. 

Results:

In GC2 cells, BOLL expression significantly increased after adding Aza-deoxycytidine with 10% FBS. In Hek293T cell, BOLL expression significantly increased after transfection of the whole BOLL sequence at different concentrations of FBS. At FBS 10%, spermatid markers, including PRM2, GAPDHS, TNP2, and CATSPER3, was significantly increased in demethylated GC2 cell compared to control. In Hek293T BOLL transfected cells, the increased spermatid markers were PRM2, GAPDHS, TNP2, and CATSPER1. Both ID4 and PIWI4 were significantly decreased in Hek293T BOLL transfected cells.

Conclusions:

Demethylation can increase BOLL expression in GC2 cells at higher concentrations of FBS. Hek293T BOLL transfected cells can effectively increase BOLL expression. Increased BOLL expression will induce a situation similar to spermatocyte cell differentiation.