Research On Proliferation Of Mouse C2C12 And Satellite Cells Regulated By Proliferin

The mechanism of skeletal muscle growth, development and regeneration, one of the hot spots in the research of animal genetics and medical science, is significant for animal muscle growth and regeneration. Proliferin(PLF), belonging to Prolactin/Growth hormone superfamily, is involved in angiogenesis, migration of endothelial cells and keratinocytes in hair follicle, and in the proliferation of tumor cells. However, the role of PLF in muscle has not been reported.Our study took mouse C2C12 myoblasts and satellite cells as models to explore expression regularity and regulatory mechanism of PLF in myoblast proliferation and differentiation, which can lay the foundation for further research of skeletal muscle growth and development. The main results were as follows:(1) The temporal and spatial expression pattern of PLF in C2C12 cells was identified, and Prl2c2 gene was confirmed as a dominant one. RT-PCR and Q-PCR results showed that PLF expression was higher in proliferating C2C12 cells than in differentiating ones(p<0.01). Digest reaction and monoclonal sequencing showed that Prl2c2 played a dominant role.(2) The biological characteristics of PLF, promoting C2C12 cell proliferation, while inhibiting cell differentiation, were identified. After C2C12 cells were transfected with si-PLF, the results of flow cytometry analysis showed that the proportion of cells was significantly increased in the G0/G1 phase(p<0.01), while decreased in the S phase(p<0.01). x CELLigence monitoring system showed that cells transfected with si-PLF had a lower growth rate. The Ed U staining experiment showed that cells transfected with si-PLF had a lower Ed U positive rate(p<0.01). After being induced to differentiate, cells transfected with si-PLF were detected by Q-PCR. It was turned out that Myo G, muscle differentiation marker, was significantly upregulated(p<0.05).(3) PLF promoted C2C12 cells proliferation through MAPK and Wnt signaling pathways.(4) The gene networks, interacting with PLF, were confirmed by si RNA and Q-PCR. Q-PCR results showed that Xirp1, Rrm2 b, Ywhaz and Pole4 m RNA were highly significantly reduced, and Ttn m RNA was significantly reduced after C2C12 cells were transfected with si-PLF.(5) The temporal and spatial expression pattern of PLF in mouse satellite cells was identified. RT-PCR results showed that PLF expression was higher in proliferating satellite cells than in differentiating ones.In summary, PLF promoted myoblasts and satellite cells proliferation, while inhibited cell differentiation. In addition, PLF played an important role in skeletal muscle growth and development through MAPK and Wnt signal transduction pathway. These findings provided a theoretical basis for elucidating the function of PLF in muscle-derived cells.