Effect Of EGFR-modified Gold Nanoparticles On Hypopharyngeal Squamous Cell Carcinoma In Nude Mice

Objective(s):Hypopharyngeal squamous cell carcinoma is one of the malignant tumors of the head and neck,which is extremely prone to metastasis,resulting in a five-year survival rate of only 30%-35%,and the prognosis is extremely unfavorable.How to inhibit or even cure hypopharyngeal carcinoma has become an increasingly concerned issue in the medical field.The aim of this study is to investigate the effect and mechanism of gold nanoparticles(AuNRs)and EGFR-m Ab/AuNRs conjugate near red on hypopharyngeal carcinoma xenografts in nude mice.Methods:1.Firstly,the subcutaneous hypopharyngeal squamous cell carcinoma xenograft model was established in nude mice.2.Different treatment methods were used to treat the transplanted tumors: equal volume of normal saline(0.9%N.S.)was injected;Injection of gold nanoparticles solution(AuNRs.);EGFR-m Ab(EGFRm Ab.)was injected;The tumors were treated with NIR laser irradiation.EGFR-m Ab/Ausolution(EGFRm Ab/AuNRs.)was injected;NIR laser irradiation(AuNRs.+NIR)was performed 24 hours after injection of aun Rs solution.NIR laser irradiation(EGFRm Ab/AuNRs.+NIR)was performed24 h after EGFR-m Ab/Ausolution injection.The drug injection was divided into tumor direct injection(it)group and tail vein injection(iv)group.The model mice were randomly divided into different groups for different treatments,and the tumor tissues were removed after treatment.3.The daily volume of tumor and the weight of tumor in vitro were recorded,and the tumor inhibition of each treatment method was calculated.The expression of tumor pyroptosis-related proteins was detected by Q-PCR and Western blotting,and SPSS26.0 and Graph Pad 8.0 were used for drawing and data analysis.P<0.05 indicated that the difference was statistically significant.Results:1.The tumor inhibition of each group was detected by tumor volume and weight.The results suggested that compared with the 0.9%NS group,all the groups except AuNRs.it group had an inhibitory effect on tumor growth from the perspective of tumor mass.In terms of volume,all the groups except the NIR group did not produce significant inhibitory effect,and no trend of large reduction in tumor volume was found.2.Through Q-PCR,we found that the expression of pyroptosis gene GSDMD was significantly different.By statistical analysis,GSDMD gene expression was more in the EGFR-m Ab/Au+NIR group compared to the other groups.Compared with tail vein injection,direct tumor injection is more effective in increasing the expression of GSDMD gene.3.By Western Blotting,we found that the expressions of the pyroptosis-promoting proteins Caspase-1,4,5,11 and GSDMD were significantly different.(1)In the detection of Caspase-1 expression,compared with other groups,the EGFR-m Ab/Au+NIR group had more Caspase-1 expression.For the expression of Caspase-1,there was no significant difference between the AuNRs+NIR group and the aun Rs +NIR group,except that the aun Rs +NIR group was more effective than the caudal vein injection.(2)The expression of Caspase-4 in EGFR-m Ab/Au+NIR group was higher than that in other groups.For the expression of Caspase-4,the direct injection of EGFRm Ab/AuNRs group and AuNRs+NIR group was relatively more effective than tail vein injection,but there was no significant difference in other groups.(3)In the detection of Caspase-5 expression,compared with other groups,the expression of Caspase-5 in EGFR-m Ab/Au+NIR group increased.However,there was no significant difference in the expression of Caspase-5 between the direct injection and the tail vein injection.(4)The expression of Caspase-11 in EGFR-m Ab/Au+NIR group was higher than that in EGFR-mab /Au+NIR group.For the expression of Caspase-11,except for the AuNRs group and the EGFRm Ab group,the direct injection of tumors was more effective than the tail vein injection.(5)Compared with other groups,the expression of GSDMD was increased in EGFR-m Ab/Au+NIR group.For the expression of GSDMD,there was no significant difference between the direct injection route and the tail vein injection route.Conclusion(s):In this study,near infrared heat of EGFR-m Ab/AuNRs conjugate on the surface could significantly inhibit the activity of hypopharyngeal carcinoma grafts in nude mice.The effect of direct tumor injection may be better than that of caudal vein injection.The mechanism of inhibiting the activity of hypopharyngeal carcinoma grafts in nude mice may be related to the up-regulation of Caspase-1,4,5,11 and GSDMD proteins,which induce cell scorch death.