Effect of CYP17A1 Gene Knockdown on Expression of Genes Related to Testosterone Synthesis and Testicular Development in Goat (Capra hircus) Leydig Cells
ZHANG Yuan1,2, CHEN Xiang1,2,*, ZHANG Yan1,2, FU Kai-Bin1,2, TANG Wen1,2, CHEN Jia-Jing1,2, JI Tao-Tao1,2, YANG Pei-Fang1,2
1 College of Animal Science, Guizhou University/Key Laboratory of Animal Geneties, Breeding and Reproduction in the Plateau Mountain Region, Ministry of Education, Guiyang 550025, China; 2 Guizhou Key Laboratory of Animal Genetics and Breeding, Guiyang 550025, China
Abstract：Cytochrome P450c17 (CYP17A1 ) is a rate-limiting enzyme in the pathway of cholesterol synthesis and plays a key role in androgen biosynthesis. It is of great significance to clarify the effect of CYP17A1 on testosterone synthesis and testicular development in goats (Capra hircus) for improving the fertility of rams. In this study, RNA interference vector of CYP17A1 was constructed and transfected into Leydig cells of Qianbei Ma goat. The knockdown effect of CYP17A1 gene was detected by qRT-PCR and Western blot. The effect of CYP17A1 knockdown on the proliferation of Leydig cells was detected by CCK-8 (Cell Counting Kit-8). The expression levels of proliferation-related genes, testosterone synthesis-related genes and testicular development-related genes were detected by qRT-PCR. The results showed that CYP17A1 was expressed in the testis and epididymis of each month old, and its gene knockdown effect was obvious, in which the expression of CYP17A1 gene and protein was significantly down-regulated (P<0.01), and the proliferation of Leydig cells was significantly inhibited (P<0.01). CYP17A1 gene knockdown significantly down-regulated the expression of proliferating cell nuclear antigen (PCNA) and cyclin E, CYP11A1, 3β-hydroxysteroid dehydrogenase type 1 (HSD3B1), HSD17B3 and CYP19A1, deleted in azoospermia like (DAZL) and insulin like growth factor 2 (IGF2)(P<0.01). This study provides basic data for further exploring the effect of CYP17A1 gene on the reproductive traits of Qianbei Ma goat.
张远, 陈祥, 张艳, 付开斌, 唐文, 陈家靖, 嵇桃桃, 杨沛方. CYP17A1基因敲低对山羊睾丸间质细胞睾酮合成和睾丸发育相关基因表达的影响[J]. 农业生物技术学报, 2023, 31(12): 2535-2544.
ZHANG Yuan, CHEN Xiang, ZHANG Yan, FU Kai-Bin, TANG Wen, CHEN Jia-Jing, JI Tao-Tao, YANG Pei-Fang. Effect of CYP17A1 Gene Knockdown on Expression of Genes Related to Testosterone Synthesis and Testicular Development in Goat (Capra hircus) Leydig Cells. 农业生物技术学报, 2023, 31(12): 2535-2544.
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