Previous studies have shown that four and a half LIM domain protein 2 (FHL2) plays an essential role in the regulation of follicular development in mammals. Although the FHL2 genes of human and mouse have been well characterized, the expression and location of FHL2 in ovary and the biological functions of FHL2 on granulosa cells (GCs) of ovine are still not clear. In this study, full‐length complementary DNA (cDNA) of FHL2 from ovine follicular GCs was amplified by real‐time PCR (RT‐PCR). The expression and location of FHL2 in ovary and GCs of ovine were studied by immunohistochemistry and immunofluorescence, and the biological effects of FHL2 on the cell proliferation, cell apoptosis, cell cycles and expression level of related genes of ovine GCs were also explored by overexpression or knockdown of FHL2. The results indicated that FHL2 was expressed in ovine follicular GCs and the sequence of the FHL2 cDNA was consistent with that predicted in GenBank, which did not cause an amino acid change. According to the results, FHL2 was expressed in ovine ovary and mainly located in the cytoplasm and nucleus of GCs. In addition, overexpression of FHL2 significantly reduced the cell viability, promoted the cell apoptosis and decreased the percentage of G0/G1 and S phase cells. RT‐PCR showed that overexpression of FHL2 significantly increased the mRNA expression level of Bax and decreased the expression of Bcl‐2 and the Bcl‐2/Bax mRNA ratio compared with the control group. Besides, the knockdown of FHL2 gene in ovine GCs significantly improved the cell viability, suppressed the cell apoptosis, decreased the mRNA expression level of Caspase‐3 gene, increased the Bcl‐2/Bax mRNA ratio and increased the percentage of S and G2/M phase cells. Our results suggest that FHL2 may play an important role in the biological functions of GCs in ovine.

中文翻译：

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四个LIM域蛋白2（FHL2）在绵羊颗粒细胞中的表达，定位和生物学作用。

先前的研究表明，四个半LIM域蛋白2（FHL2）在调节哺乳动物的卵泡发育中起着至关重要的作用。尽管已经很好地表征了人和小鼠的FHL2基因，但仍不清楚FHL2在卵巢中的表达和定位以及FHL2对绵羊颗粒细胞（GCs）的生物学功能。在这项研究中，通过实时PCR（RT-PCR）扩增了来自卵泡GC的FHL2的全长互补DNA（cDNA）。通过免疫组织化学和免疫荧光技术研究了FHL2在绵羊卵巢和胃GC中的表达和定位，并通过过度表达或FLC2研究了FHL2对绵羊GC细胞增殖，细胞凋亡，细胞周期及相关基因表达水平的生物学影响。击倒FHL2。结果表明FHL2在绵羊卵泡GC中表达，FHL2 cDNA的序列与GenBank预测的一致，没有引起氨基酸变化。根据结果​​，FHL2在绵羊卵巢中表达，并且主要位于GC的细胞质和细胞核中。此外，FHL2的过表达显着降低了细胞活力，促进了细胞凋亡并降低了G0 / G1和S期细胞的百分比。RT‐PCR显示FHL2的过表达显着增加了FHL2的mRNA表达水平 FHL2的过表达显着降低了细胞活力，促进了细胞凋亡并降低了G0 / G1和S期细胞的百分比。RT‐PCR显示FHL2的过表达显着增加了FHL2的mRNA表达水平 FHL2的过表达显着降低了细胞活力，促进了细胞凋亡并降低了G0 / G1和S期细胞的百分比。RT‐PCR显示FHL2的过表达显着增加了FHL2的mRNA表达水平Bax表达和减少的表达的Bcl-2和Bcl-2的/ Bax蛋白与对照组相比，mRNA比值。此外，绵羊GC中FHL2基因的敲低显着提高了细胞活力，抑制了细胞凋亡，降低了Caspase-3基因的mRNA表达水平，增加了Bcl-2 / Bax mRNA的比率，并增加了S和G2 /的百分比M期细胞。我们的结果表明FHL2可能在绵羊GC中的生物学功能中起重要作用。