Acta Veterinaria et Zootechnica Sinica ›› 2025, Vol. 56 ›› Issue (2): 582-590.doi: 10.11843/j.issn.0366-6964.2025.02.010

• Animal Genetics and Breeding • Previous Articles     Next Articles

miR-665 Targeting BCL2L11 Regulates the Myoblasts Proliferation of Wu'an Goat

FENG Jing1,2,3(), SHENG Hui1,2,3, ZHANG Xiaosheng1,2,3, GUO Xiaofei1,2,3, YAO Dawei1,2,3, LI Yupeng1,2,3, CHEN Longbin1,2,3, ZHANG Jinlong1,2,3,*()   

  1. 1. Institute of Animal Science and Veterinary, Tianjin Academy of Agricultural Sciences, Tianjin 300381, China
    2. Tianjin Key Laboratory of Animal Molecular Breeding and Biotechnology, Tianjin 300381, China
    3. Tianjin Engineering Research Center of Animal Healthy Farming, Tianjin 300381, China
  • Received:2024-08-09 Online:2025-02-23 Published:2025-02-26
  • Contact: ZHANG Jinlong E-mail:fengjing_2024@126.com;jlzhang1010@163.com

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Abstract:

This study aimed to use molecular biological methods to explore the effect of miR-665 targeting BCL2L11 on the myoblasts proliferation of goat. Three healthy male Wu'an goats aged 1 month and 9 months each, with similar body weights and feeding environments for each age group was selected.The expression of BCL2L11 and miRNA-665 in the longest dorsal muscle tissue of Wu'an goats at different ages was detected using RT-qPCR; The expression of BCL2L11 in different tissue of Wu'an goats was also detected by RT-qPCR; Wild-type and mutant vectors of BCL2L11 3'UTR, as well as miR-665 mimics and control groups, were constructed and cotransfected into HEK293T cells. The targeting relationship between miR-665 and BCL2L11 was identified using a dual luciferase reporter gene detection system. The goat myoblasts were transfected with miR-665 mimic and negative control, and the expression levels of apoptosis markers factors XIAP and Fas were detected by RT-qPCR, and the effect of miR-665 overexpression on the proliferation of goat myoblasts was analyzed by EdU test simultaneously. The results showed that BCL2L11 was highly expressed in the longissimus dorsi muscle tissue; There were differences in the expression of miR-665 and BCL2L11 in the longissimus dorsi muscle tissue of goats aged 9-months and 1-months, and the expression of miR-665 at 1-month was higher than that at 9-month, while the expression of BCL2L11 was opposite, and the two were negative regulated; The dual luciferase report analysis showed that, after transfecting overexpressed miR-665 mimic, the luciferase activity of BCL2L11 was significantly inhibited, and the mRNA expression level of the target gene BCL2L11 was also significantly reduced. In order to further verify the function of miR-665, the expression of apoptosis marker genes XIAP and Fas was also significantly reduced after transfection of miR-665 mimic; EdU test analysis showed that the proliferation of myoblasts increased significantly after overexpression of miR-665. To sum up, miR-665 as a regulatory element of BCL2L11 in goat longissimus dorsi muscle tissue, could significantly inhibit the expression level of BCL2L11 in goat myoblasts, when miR-665 was overexpressed, it could promote the proliferation of myoblasts.

Key words: miR-665, BCL2L11, muscle, Wu'an goat, myoblast

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