| Summary: | 碩士 === 國立成功大學 === 生命科學系碩博士班 === 100 === Palladin, a cytoskeleton-associated protein, plays an important role in regulating cell adhesion and the formation of stress fibers. Palladin has three major isoforms: 200kDa, 140kDa, 90~92 kDa, and the expression of different isoforms has a tissue specific feature,suggesting that different palladin isoforms may be specialized for different functions.Palladin is important in embryonic development, palladin knockout mice caused embryonic lethal, but its role in skeletal muscle development is still unclear. In my dissertation, I tried to futher analyze the role of palladin in myoblast differentiation by using small interfering RNA (siRNA) to knockdown all palladin isoforms in C2C12 myoblasts. The transfection of palladin siRNAs successfully decreased the palladin expression in the C2C12 cells. I also found knockdown of palladin decreased cell density and delayed the formation of myotubes during myoblast differentiation. The result indicated that knockdown of palladin may induce cell death or decrease cell proliferation. The MTT assay and the BrdU incorporation assay results showed that cell proliferation rate was decreased by knockdown of palladin in C2C12. The down- regulation of palladin also induced cell apoptosis by measuring caspase3/7 activity in C2C12 cells. The real-time PCR results showed that all palladin isoforms were down-regulated after transfecting with anti-pan-palladin siRNA#1. The down-regulation of myogenin in palladin knockdown C2C12 cells indicated the myoblast differentiation was affected. The expression of the proliferation marker, Ki67, was increased in palladin-knockdown C2C12 cells after inducing to differentiate. In conclusion, this study demonstrated that knockdown of all palladin isoforms in C2C12 myoblast cells can increase the apoptosis and decrease cell proliferation rate.
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