scRNA-seq reveals that origin recognition complex subunit 6 regulates mouse spermatogonial cell proliferation and apoptosis via activation of Wnt/β-catenin signaling  

作　　者：Shi-Wei Liu Jia-Qiang Luo Liang-Yu Zhao Ning-Jing Ou Chao-Yang Yu-Xiang Zhang Hao-Wei Bai Hong-Fang Sun Jian-Xiong Zhang Chen-Cheng Yao Peng Li Ru-Hui Tian Zheng Li Zi-Jue Zhu 

机构地区：[1]Department of Andrology,Center for Men's Health,Urologic Medical Center,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China [2]State Key Lab of Reproductive Medicine,Nanjing Medical University,Nanjing 211166,China [3]Shanghai Key Lab of Reproductive Medicine,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China [4]Department of Urology,Guangdong Provincial Key Laboratory of Biomedical Imaging,The Fifth Affiliated Hospital,Sun Yat-sen University,Zhuhai 519000,China [5]Department of Urology,First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China

出　　处：《Asian Journal of Andrology》2024年第1期46-56,共11页亚洲男性学杂志（英文版）

基　　金：This work was supported by the National Key Research and Development Program of China(No.2022YFC2702700);the National Natural Science Foundation of China(No.82171597);Clinical Research Plan of Shanghai Hospital Development Center(No.SHDC2020CR3077B).

摘　　要：The regulation of spermatogonial proliferation and apoptosis is of great significance for maintaining spermatogenesis.The single-cell RNA sequencing(scRNA-seq)analysis of the testis was performed to identify genes upregulated in spermatogonia.Using scRNAseq analysis,we identified the spermatogonia upregulated gene origin recognition complex subunit 6(Orc6),which is involved in DNA replication and cell cycle regulation;its protein expression in the human and mouse testis was detected by western blot and immunofluorescence.To explore the potential function of Orc6 in spermatogonia,the C18-4 cell line was transfected with control or Orc6 siRNA.Subsequently,5-ethynyl-2-deoxyuridine(EdU)and terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)assays,flow cytometry,and western blot were used to evaluate its effects on proliferation and apoptosis.It was revealed that ORC6 could promote proliferation and inhibit apoptosis of C18-4 cells.Bulk RNA sequencing and bioinformatics analysis indicated that Orc6 was involved in the activation of wingless/integrated(Wnt)/β-catenin signaling.Western blot revealed that the expression ofβ-catenin protein and its phosphorylation(Ser675)were significantly decreased when silencing the expression of ORC6.Our findings indicated that Orc6 was upregulated in spermatogonia,whereby it regulated proliferation and apoptosis by activating Wnt/β-catenin signaling.

关 键 词：cell proliferation ORCG scRNA-seq analysis SPERMATOGONIA Wnt/β-catenin signaling 

分 类 号：R697[医药卫生—泌尿科学]