Cloning of a new testis-enriched gene C4orf22 and its role in cell cycle and apoptosis in mouse spermatogenic cells
Cloning of a new testis-enriched gene C4orf22 and its role in cell cycle and apoptosis in mouse spermatogenic cells
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新睾丸富集基因C4orf22的克隆及其在小鼠生精细胞细胞周期和凋亡中的作用
DOI：
10.1007/s11033-019-04651-8
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发表时间：
2019-04-01
影响因子：
2.8
通讯作者：
Nie, Dongsong
中科院分区：
文献类型：
--
作者：
Li, Hui;Dai, Yanfa;Nie, Dongsong
精子发生是一个复杂的、动态的细胞分化过程，主要受基因、类固醇激素和环境因素的调控。虽然已经确定了一些与精子发生有关的基因，但仍有许多与精子发生有关的基因尚未得到解释。在这里，一种新的基因C4orf22，也被称为1700007G11Rik或Cfap299，从小鼠睾丸中鉴定出来。C4orf22蛋白含有233个氨基酸残基，在后生动物中高度保守。通过RT-PCR和qRT-PCR分析，C4orf22 mRNA主要在小鼠睾丸中表达，在发育过程中，C4orf22 mRNA在2周龄至8周龄的小鼠睾丸中表达增加。免疫组化分析显示，C4orf22蛋白主要分布在精原细胞和原代精母细胞的细胞质中，GC-1和GC-2细胞中标记GFP的C4orf22进一步证实了这一点。过表达pEGFP-C3-C4orf22可显著抑制GC-1细胞凋亡，促进细胞周期进程，S期和G2期细胞数量增加。相反，小干扰RNA （siRNA）沉默GC-1细胞中的C4orf22可导致凋亡细胞数量增加，细胞周期停留在G2/M期。Western blot分析和qRT-PCR结果显示，C4orf22过表达显著增加抗凋亡bcl-2的表达，降低caspase-3、caspase-8和Bax的表达。我们的研究结果提示C4orf22可能参与精子发生，并首次揭示了其在GC-1细胞中通过bcl-2调控通路调控细胞凋亡的潜在作用。
Spermatogenesis is a complicated and dynamic cellular differentiation process mainly regulated by genes, steroid hormones and environmental factors. Although a number of genes involved in spermatogenesis have been identified, there are still a lot of genes underlying spermatogenesis remained unexplained. Here, a novel gene C4orf22, also known as 1700007G11Rik or Cfap299 was identified from mouse testis. C4orf22 protein contains 233 amino acid residues and is highly conserved in metazoan species. C4orf22 mRNA was predominantly expressed in mouse testis and increased from 2-week-old testes to 8-week-old testes during the developing testes by RT-PCR and qRT-PCR. Immunohistochemical analysis indicated that C4orf22 protein was mainly distributed in the cytoplasm of spermatogonia and primary spermatocytes, which was further confirmed by C4orf22-tagged with GFP in the GC-1 and GC-2 cells. Over-expression of pEGFP-C3-C4orf22 significantly inhibited GC-1 cells apoptosis and promoted cell cycle progression with an increase in the cell number of S and G2 phase. Conversely, small interfering RNA (siRNA) silencing C4orf22 in GC-1 cells could cause an increase in the number of apoptosis cells and the cell cycle was arrested at G2/M phase. Western blot analysis and qRT-PCR results showed that C4orf22 over-expression significantly increased the expressions of anti-apoptotic bcl-2 and decreased the expression of caspase-3, caspase-8 and Bax. Our results suggest that C4orf22 may be involved in spermatogenesis, and for the first time, unravels its potential role in regulating cell apoptosis through bcl-2 regulatory pathway in GC-1 cells.