Effect And Mechanism Of Sevoflurane On The Growth,Invasion And Metastasis Of Osteosarcoma Cells

Objective(s):Osteosarcoma is one of the common malignancies in orthopedics.Most patients are young and have poor prognosis.How to improve the prognosis of patients is a major problem.Surgical resection as the main treatment means that each step in the surgery will have a different impact on the treatment of osteosarcoma.Sevoflurane is widely used for general anesthesia because it has good analgesia,good controllability,and easy patient tolerance.More and more evidence shows that sevoflurane shows different effects on different tumor cells.The proliferation and invasion of tumor cells are the key links in tumor metastasis.It is necessary to understand the effect of sevoflurane on the proliferation and invasion of tumor cells.The effect of sevoflurane on tumors is a hot spot in current research.There have been many reports on the effects of sevoflurane anesthesia on some tumors such as lung cancer and breast cancer.The study found that genes P53,MMP2 play an important role in the occurrence and metastasis of tumors,TGF? gene also has an impact on the formation of osteosarcoma.However,the effect of sevoflurane on osteosarcoma has rarely been reported.With the increase in the proportion of sevoflurane general anesthesia in patients with osteosarcoma,it is more meaningful to study the effect of sevoflurane on osteosarcoma cells.This article aims to study effects of the same time on proliferation,apoptosis,migration and invasion of osteosarcoma Saos2 cells with the treatment of different concentrations of sevoflurane(1.7%sevoflurane,3.4%sevoflurane,5.1%sevoflurane)by MTT cell proliferation assay,flow cytometry assay,cell scratch assay and Transwell invasion assay.Different concentrations of sevoflurane on P53,MMP2,and TGFp were studied by RT-PCR and Western blot experiments.The effect of gene expression in Saos2 cells hopes to make sevoflurane more rational and scientifically applicable to patients with osteosarcoma.Methods:Saos2 cells were purchased,cultured,passaged,cryopreserved,resuscitated,and seeded on different cell culture plates or vials for culture according to experimental requirements.Logarithmic phase Saos2 cells were seeded into 96-well plates,and Saos2 cells adherently grown with different concentrations of sevoflurane were treated for 2 h and divided into 1.7%sevoflurane group(s1)and 3.4%sevoflurane group(s2).5.1%sevoflurane group(s3),control group(c).Each group was subjected to MTT cell proliferation experiments,and SPSS software was used to analyze and calculate the inhibition rate of each group.Logarithmic growth of Saos2 cells was inoculated into six-well plates.Fluorescent dyes were added after treatment with different concentrations of sevoflurane.Flow cytometry was used to determine the apoptotic rate of Saos2 cells in each group to observe the apoptosis of Saos2 cells induced by sevoflurane.The effect of logarithmic growth of Saos2 cells inoculated into six-well plates,different concentrations of sevoflurane treatment,through the cell scratch test to understand the migration of each group,to study the effects of sevoflurane on Saos2 cell migration;Logarithmic growth of Saos2 cells was inoculated into 24-well plates.After treatment with different concentrations of sevoflurane,the invasiveness of each group was investigated by Transwell invasion experiment to study the effect of sevoflurane on Saos2 cell invasiveness;Logarithmic growth was used.Saos2 cells were inoculated into cell culture vials and treated with different concentrations of sevoflurane.Proteins and gene expression of P53,MMP2,TGF? in Saos2 cells were detected by Western blotting and RT-PCR experiments.The effect of sevoflurane on P53,MMP2 and TGF?was observed.Results:Compared with the control group and different concentrations of sevoflurane group,low concentration(? 1.7%)sevoflurane inhibited Saos2 cell proliferation,high concentration(? 3.4%)sevoflurane showed a role in promoting Saos2 cell proliferation,the difference has statistics Significance(P<0.05);The proportion of apoptotic Saos2 cells in the sevoflurane group was higher than that of the control group.Sevoflurane could promote the apoptosis of Saos2 cells;Compared with the control group,the sevoflurane group's ability to migrate significantly decreased.In the Transwell invasion assay,Saos2 cells showed decreased invasive ability compared with the test group.By RT-PCR experiments,the target band of P53 gene was not detected in agarose gel electrophoresis analysis,and the target band of MMP2 gene was clearly detected on the image.By Western blotting experiments,no target band was found in the image exposure of gene P53;in the Western blot image of MMP2 gene,the target band of sevoflurane treatment group was increased compared with the control group;Western blot of TGF-? gene was performed.In the image,the band of the target gene increases after sevoflurane treatment.Conclusion(s):Sevoflurane has the effect of promoting or inhibiting the proliferation,migration and apoptosis of Saos2 osteosarcoma cells,and is related to the concentration.Low concentrations(?1.7%)of sevoflurane inhibit Saos2 cell proliferation,high concentrations(?3.4%)of sevoflurane promote Saos2 cell proliferation,sevoflurane promotes Saos2 cell apoptosis,inhibits Saos2 cell migration,and promotes Saos2 cells Invasion ability.In Saos2 cells,there is no expression of the P53 gene.Sevoflurane promotes TGF-? gene expression in Saos2 and inhibits MMP2 gene protein expression.