Department of Clinical laboratory, Children's Hospital of Soochow University, Suzhou, P. R. China.
Section of Pediatric Urology, Children's Hospital of Soochow University, Suzhou, P. R. China.
Birth Defects Res. 2021 Mar 15;113(5):399-408. doi: 10.1002/bdr2.1863. Epub 2021 Jan 15.
Hypospadias is a common birth defect that might be caused by inadequate fusion of the urethral folds in the process of male external genital development. We intended to discover the crucial long noncoding RNAs (lncRNAs) regulating autophagy from the gene expression profile of the genital tubercle (GT) of dibutyl phthalate (DBP) induced hypospadiac rats.
Whole transcriptome resequencing was used to determine the expression of the total RNA in GTs and cultured fibroblasts obtained from GTs of DBP-induced hypospadiac male rat fetuses. Autophagosomes and autolysosomes were examined under a transmission electron microscope after overexpression of lncRNA NONRATT008453.2 in the fibroblasts by adenovirus transfection. Finally, the protein expression levels of Atg5, Beclin-1, Atg7, and the LC3A/B-II:LC3A/B-I ratio were detected in the fibroblasts by western blotting.
NONRATT008453.2 suppressed autophagy by promoting the expression of Atg7, but inhibited the expressions of Atg5, Beclin-1, and the LC3A/B-II:LC3A/B-I ratio in the GT fibroblasts.
NONRATT008453.2 may have an influence on autophagy in the fibroblasts of the GT in DBP-induced hypospadiac rats.
尿道下裂是一种常见的出生缺陷，可能是男性外生殖器发育过程中尿道褶融合不全引起的。我们旨在从邻苯二甲酸二丁酯（DBP）诱导的尿道下裂大鼠生殖结节（GT）的基因表达谱中发现调节自噬的关键长非编码 RNA（lncRNA）。
全转录组重测序用于确定 GT 和从 DBP 诱导的尿道下裂雄性胎鼠 GT 中分离的成纤维细胞中总 RNA 的表达。用腺病毒转染使成纤维细胞中 lncRNA NONRATT008453.2 过表达后，在透射电子显微镜下观察自噬体和自溶酶体。最后，通过 Western blot 检测成纤维细胞中 Atg5、Beclin-1、Atg7 和 LC3A/B-II：LC3A/B-I 比值的蛋白表达水平。
NONRATT008453.2 通过促进 Atg7 的表达抑制自噬，但抑制 GT 成纤维细胞中 Atg5、Beclin-1 和 LC3A/B-II：LC3A/B-I 比值的表达。
NONRATT008453.2 可能对 DBP 诱导的尿道下裂大鼠 GT 成纤维细胞中的自噬有影响。
