A Study Of PI3K/AKT Signal Pathway In The Antler Stem Cells

In recent years, studies show that the PI3K/AKT signaling pathway is closely related withtumorigenesis, the regulating mechanism is becoming a breakthrough point in studying cancer.PI3K/AKT signaling pathway has a role in the regulation of cell cycle and proliferation, inhibition ofapoptosis, promotion of cell invasion and migration. Deer antlers a new research model for thetreatment of tumors are renewed in yearly cycles, whose regeneration are based on the proliferation anddifferentiation of cells. The stem cells of deer antlers are proliferating rapidly but not showing cancerous,which has not yet clearly revealed the regulate mechanism.In this paper, we studied the affection of antler stem cells’ physiology when PI3K/AKT signalingpathway is inhibited or up-regulated by LY294002or insulin. We use Antlerogenic periosteumcells(APCs), pedicle periosteum cells(PPCs) and Face periosteum cells(FPCs) as the experimentalsubjects.(1) To detect cells’ proliferate inhibition rate at the time of12h,24h,48h by MTT staining whenLY294002’s concentration was0,5,15,25,35,50;(2) To observe cytoskeleton staining with CoomassieBrilliant Blue R250in the above-mentioned concentration gradient;(3) To detect cells’ adhesion in the96-well plates treated by Matrigel basement membrane(low growth factor)when AP, PP and FP cells arecultured in presence of insulin(10-6mol/L) or LY294002(50μmol/L);(4) To detect cell cycle with flowcytometry when AP, PP and FP cells are cultured with different stimulate factors.The results showed that (1) inhibition rate of three types of cells’ proliferation increased with theincrease in time and inhibitor concentration. The difference on the inhibition rate reflects that the degreeof regulation of the PI3K/AKT signaling pathway is not the same, which is worthy of further study.(2)AP and PP cells’ cytoskeleton were influenced obviously by insulin and LY294002, which can reflectthat PI3K/AKT signaling pathway has an important role in the AP and PP cells; on the contrary, FP cells’result was not so obvious.(3)AP and PP cells’ adhesion rate were significantly in the process of thedifferent conditions; The results of FP cells between the experimental group and the control group wasnot significantly different, but there was a significant difference compared with the insulin experimentalgroup.(4)PP1/3cells cycle was absolutely changed between experimental group and control group, butthe same with PP2/3cell cycle, there was no significantly difference for AP and PP cells in each group.This experiment was set up the PI3K/AKT signaling path as the object of study, measured cellphysiological characteristics and verified the position of PI3K/AKT signaling pathway in the antler stemcells. At the same time, this experiment provides a reference for studying subsequent downstreamfactors and fills the research gaps in the process of antler regeneration of cell physiologicalcharacteristics, and has important implication for the study of the antler regeneration and treatment ofcancer.