Research Of Low-dose Extracorporeal Shock Wave Through ATP Assisted With Methotrexate On Apoptosis In Osteosarcoma U2OS Cells

Low doses extracorporeal shock wave can increase the permeability of the cellmembrane and induce extracellular macromolecules into cells. But can also cause the releaseof intracellular ATP extracellularly. To release high concentrations of extracellular ATPcauses changes in cell function. In vitro experiments showed that extracorporeal shock wavesynergistically with chemotherapy drugs can induce apoptosis.This suggests thatextracorporeal shock wave can be used to assist in the treatment of cancer chemotherapeuticdrugs. However, these studies lack vitro experiments to support. Ignoring the fact that theextracorporeal shock wave can cause the release of intracellular ATP to the extracellular.Based on this, we assume that Extracorporeal shock wave by varying the permeability ofthe cell membrane, on the one hand directed to the increase in intracellular ATP is releasedoutside the cell by cell surface receptors P2X7, osteosarcoma promote apoptosis; the otherhand, the chemotherapy drug into the targeted increase in the intracellular content, so thatyou can to reduce the amount of chemotherapeutic drugs, while increasing apoptosis inosteosarcoma purpose. Thus this study will be how to increase the sensitivity of osteosarcomacells to chemotherapeutic drugs, reducing drug dosage of chemotherapy, targeted therapy ofsolid tumors to enhance the effect, providing a new means and theoretical basis.Objective Studies of extracorporeal shock wave synergistic low doses of methotrexateinduce mechanism of human osteosarcoma U2OS cells apoptosis. Study whether low-doseextracorporeal shock wave caused by an increase in the permeability of cell membranes.Onthe one hand to increase which is not easily through the cell membrane chemotherapydrug-methotrexate into the cell membrane and enhance apoptosis; on the other hand,topromote releasing ATP to the extracellular, activating the P2X7receptor of cell surface, byinternal or external means apoptosis pathway to promote apoptosis in osteosarcoma cells.Methods This study culture osteosarcoma U2OS cells in vitro, using differentfrequency, capacitance0.3μF, operating voltage7kV of LDESW effect U2OS cells and detecting on the cell activity affecting LDESW by trypan blue staining.Differentconcentrations of MTX were applied and effects the United LDESW U2OS cells24h afterapplication of CCK-8assay monotherapy group and the combination group on U2OS cellproliferation inhibition.Different concentrations of MTX alone and combined LDESW impacton U2OS cell apoptosis by flow cytometry; this study, ELISA was used to detect single-druggroup and the joint group U2OS cells MTX content;Finally,we quantitatively analyzed thecontent of U2OS extracellular ATP in the control group and the experimental group byHPLC.Results The experimental study showed that: capacitance0.3μF, operating voltage7KV of LDESW effects on U2OS cells less than300times does not affect cell viability.Monotherapy group compared to the combination group significantly inhibit cell proliferationand induce apoptosis (P<0.05).ELISA was used to detect single-drug group than in thecombination group compared to U2OS cells MTX content is low, compared to the two groupsthere was a significant difference (P<0.05).HPLC detection in U2OS extracellular ATP levels,the combined group U2OS cells outside the ATP content higher,compared to the two groupsthere was a significant difference (P<0.05).Conclusion This study demonstrated that LDESW U2OS osteosarcoma can transientlyincreases the permeability of the cell membrane, on the one hand by the MTX is not easy toenter the cell membrane; on the other hand that the release of intracellular ATP to the cell, thehigh concentration of extracellular ATP formation.But did not confirm whether the highconcentrations of extracellular ATP is coordinated through the activation of P2X7receptorpathway MTX-induced apoptosis in osteosarcoma cells. Further study need follow-up tests.