| Objective: To investigate the effect of scorpion venomous polypeptide(PESV)on the activation pathway CIRC?0016760 /HIF-1 ? in non-small cell lung cancer(NSCLC)and its effect on the growth,proliferation and invasion of NSCLC.Methods:1.Human lung cancer A549 cells,H1299 cells and human normal bronchial epithelial 16 HBE cell models were prepared and the expression levels of Circ?0016760(Circ?0016760)were detected by real-time quantitative reverse transcription polymerase chain reaction(q RT-PCR).The expression level of Circ?0016760 was determined by q RT-PCR in two lung cancer cells treated with high,medium and low doses of PESV and blank control,respectively.2.The blank control group,hypoxia treatment group,Circ?0016760 group combined with hypoxia treatment and other circ RNA knockout groups were set up in the two types of lung cancer cells respectively;The expression level of Circ?0016760 was determined by q RT-PCR.Transwell invasion assay was used to test the ability of cell migration and invasion.Cell colony formation and MTT assay were used to determine cell proliferation ability.3.The blank control group,hypoxia treatment group,hypoxia combined with PESV treatment group,hypoxia combined with PESV treatment and transfected with other circ RNA groups and hypoxia combined with PESV treatment and transfected with Circ?0016760 group were set up in the two kinds of lung cancer cells respectively,and the expression level of Circ?0016760 was measured again.Transwell assay,colony formation and MTT assay were used to determine the ability of cell migration,invasion and proliferation.4.One of the two types of lung cancer cells with high expression of CIRC?0016760 was selected to make subcutaneous xenograft tumor model of human lung cancer nude mice.A total of 24 male BALB/c nude mice,aged 5 weeks,were randomly divided into 4 groups.They were blank control group,Circ?0016760 knocked out group,PESV treatment group and Circ?0016760 overexpressed combined with PESV treatment group,respectively.A total of 28 days were observed.Finally,the volume of transplanted tumor in the 4 groups was observed and the weight of transplanted tumor was measured.The expression of CIRC?0016760 and HIF-1? were determined by q RT-PCR and W-B method.Results:1.The expression of CIRC?0016760 in tissues and cells of non-small cell lung cancer(NSCLC)was significantly higher than that in normal bronchial epithelium(P<0.05).The expression of CIRC?0016760 in NSCLC group was significantly down-regulated after PESV treatment(P<0.05).2.The migration,invasion and proliferation of NSCLC cells were significantly increased in the hypoxia group,which was statistically significant compared with the normoxia group(P<0.05).The migration,invasion and proliferation of NSCLC cells in Circ?0016760 knockout group were significantly decreased compared with the control group(P<0.05).3.The migration,invasion and proliferation of NSCLC cells in the PESV group were significantly lower than those in the control group,with statistical significance(P<0.05).The over-expression of Circ?0016760group showed that the effect of PESV treatment on NSCLC under hypoxia condition was significantly inhibited,which was statistically significant compared with the over-expression of PCD5-CIR group(P<0.05).4.The expression of CIRC?0016760 and HIF-1? in PESV group were significantly decreased compared with the control group(P<0.05).Compared with the control group,the volume and weight of subcutaneous tumor transplantation in nude mice were lower(P<0.05).The expression of HIF-1? in Circ?0016760 group was decreased,and the volume and weight of subcutaneous tumor transplantation in nude mice were lower,which was statistically significant compared with the control group(P<0.05).The expression of HIF-1? in Circ?0016760 group combined with PESV was higher than that in PESV group alone,and the volume and weight of subcutaneous xenograft in nude mice were increased,with statistical significance(P<0.05).Conclusions: The expression of CIRC?0016760 and HIF-1? were significantly down-regulated after PESV treatment,and the migration,invasion and proliferation ability of NSCLC cells were decreased,which inhibited the growth of subcutaneous xenograft of human lung cancer in nude mice.Overexpression of Circ?0016760 increased the migration,invasion and proliferation of NSCLC cells,and partially reversed the inhibitory effect of PESV on the migration,invasion and proliferation of NSCLC cells.It is suggested that Circ?0016760 may be an important central link in the treatment of NSCLC by PESV.This finding provides a possible molecular basis for the treatment of NSCLC by PESV. |
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