Effect of BCAS2 Gene on Apoptosis of Porcine (Sus scrofa) Granulosa Cells
LI Ming-Na1,*, LI Ying-Ying1, SUN Jing-Hua1, QIN Shi-Zhen1, ZHU Cai-Ye1, WANG Xin-Rong1, ZHAO Fang-Fang1, YANG Yao2
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 Gannan Animal Disease Prevention and Control Center, Gannan 747000, China
Abstract：Breast carcinoma amplified sequence 2 (BCAS2) can participate in DNA damage responses, protein stability changes and pre-mRNA splicing processes in a variety of cells. There are many studies on the role of BCAS2 gene in germ cell formation, early embryonic development, and ovarian granulosa cells in Mus musculus, but whether this gene is involved in the regulation of granulosa cell proliferation or apoptosis in livestock follicles has not been reported. In this study, follicle stimulating hormone (FSH) was added to porcine granulosa cells cultured in vitro to observe whether it affected the expression of BCAS2; then BCAS2 was knocked down using RNAi technology to detect cell apoptosis and the expression levels of related genes. The results showed that both BCAS2 mRNA and protein expression were significantly increased after 24 h of treatment with 100 ng/mL FSH (P<0.05). After BCAS2 knockdown, the apoptosis rate of granulosa cells was significantly increased (P<0.05), the expression of B-cell lymphoma-2 gene (Bcl-2) was significantly decreased (P<0.05), the expression of proliferating cell nuclear antigen gene (PCNA) was extremely significantly decreased (P<0.01), and the expression level of BCL2-associated X protein gene (Bax) was significantly increased (P<0.05). In addition, the mRNA expression of tubulin β gene (TUBB), TUBA1A and estrogen receptor 2 gene (ESR2) were extremely significantly decreased (P<0.01), ESR1 expression was significantly decreased (P<0.05), and tumor protein p53 gene (p53) expression was significantly increased (P<0.05). It showed that BCAS2 gene knockdown can induce granulosa cells apoptosis in Sus scrofa, and the regulatory mechanism may be involved in the possible pre-mRNA splicing, estrogen receptor (ER) transcriptional regulation, or p53 pathways. This study provides a theoretical basis for further revealing the potential regulatory mechanism of follicular development in domestic animals.
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