Abstract
CircRNA is a type of closed circular non-coding RNA formed by reverse splicing and plays an important role in regulating the growth and development of plants and animals. To investigate the function of circ-FoxO3 in mouse myoblast cells’ (C2C12) differentiation and proliferation, we used RT-qPCR to detect the expression level of circ-FoxO3 in mouse myoblast cells at different densities and different differentiation stages, and the specific interference fragment was used to inhibit the expression level of circ-FoxO3 in myoblast cells to observe its effect on myoblast cells proliferation and differentiation. We found that the expression level of circ-FoxO3 in myoblast cells increased with the prolongation of myoblast cells differentiation time, and its expression level decreased with the proliferation of myoblast cells. At the same time, we found that the differentiation ability of the cells was significantly increased (p < 0.05), but the cell proliferation was unchanged (p > 0.05) after inhibiting the expression of circ-FoxO3 in myoblast cells. Combining the results of bioinformatics analysis and the dual luciferase reporter experiment, we found that circ-FoxO3 is a sponge of miR-138-5p, which regulates muscle differentiation. Our study shows that circ-FoxO3 can inhibit the differentiation of C2C12 myoblast cells and lay a scientific foundation for further study of skeletal muscle development at circRNA levels.
Highlights
Skeletal muscle is one of the most important components of animals, is involved in exercise and energy metabolism, and is directly related to livestock growth and meat production [1,2]
Our study reveals that silencing circ-foxO3 promotes myoblast cells (C2C12) differentiation, which further reveals the regulation mechanism of muscle development and provides a new perspective for increasing muscle mass
The circRNA junction site sequence of circ-FoxO3 was verified by RT-polymerase chain reaction (PCR)
Summary
Skeletal muscle is one of the most important components of animals (constitutes 40% to 50% of the animal body), is involved in exercise and energy metabolism, and is directly related to livestock growth and meat production [1,2]. Myogenesis is the process by which a somatic cell undergoes a series of proliferation, migration, and differentiation to form muscle tissue during development of the embryo [9,10]. Recent studies have shown that circular RNAs (circRNAs) have been identified as emerging non-coding RNAs during skeletal muscle development and play a crucial role in muscle development [11,12,13,14,15,16,17,18,19], but the specific functions in the proliferation and differentiation of myoblast cells are still unclear. CircRNAs are non-coding RNA molecules ubiquitous in eukaryotes that are formed by reverse splicing by non-canonical splicing [20].
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