Effect Of EGFR-mAb/AuNRs Targeted Photothermal Induces Pyroptosis Of Hypopharyngeal Cancer Cells In Nude Mice

Objective(s):Currently,the treatment of hypopharyngeal cancer remains a challenge for clinicians.This study aims to explore the effect of EGFR-mAb/AuNRs targeted photothermal therapy on the occurrence of pyroptosis in nude mice with hypopharyngeal cancer xenografts,with a view to providing new ideas for the treatment of hypopharyngeal cancer.Methods:1.The human hypopharyngeal squamous cell carcinoma Fadu cell line was inoculated subcutaneously into the back of nude mice to construct a subcutaneous transplanted tumor model of hypopharyngeal carcinoma in nude mice and then these model mice were randomly divided into 13 groups for different treatments,respectively:(1)Intratumoral injection of the same volume of normal saline group(0.9% NS.it group);(2)In the group injected with an equal volume of saline via the tail vein(0.9% NS.iv group);(3)The tumor body was directly injected with gold nanorod solution group(AuNRs.it group);(4)Tail vein injection of nanogold rod solution group(AuNRs.iv group);(5)The tumor body was directly injected with anti epidermal growth factor receptor monoclonal antibody group(EGFRmAb.it group);(6)Tail vein injection of anti epidermal growth factor receptor monoclonal antibody group(EGFRmAb.iv group);(7)Near infrared laser irradiation group only for local tumor body(NIR group);(8)Intratumoral injection of epidermal growth factor receptor monoclonal antibody modified gold solution group(EGFRmAb/Au.it group);(9)Tail vein injection of epidermal growth factor receptor monoclonal antibody modified gold solution group(EGFRmAb/Au.iv group);(10)NIR laser irradiation group of tumor site after 24 hours of direct injection of nano-gold rod solution into tumor body(AuNRs.it+NIR group);(11)NIR laser irradiation group at tumor site 24 hours after Caudal vein injection of nano-gold rod solution(AuNRs.iv+NIR group);(12)NIR laser irradiation group of tumor site after 24 hours of direct injection of nano-gold solution modified by epidermal growth factor receptor monoclonal antibody(EGFR-mAb/Au.it+NIR group);(13)NIR laser irradiation group at tumorsite 24 hours after Caudal vein injection of nano-gold solution modified by epidermal growth factor receptor monoclonal antibody(EGFR-mAb/Au.iv+NIR group).The tumor tissue was removed after treatment;2.The tumor tissue was prepared into cell suspension,and the cell morphology of different treatment groups was observed by scanning electron microscope;3.The tumor tissues of different treatment groups were prepared into pathological sections.The expression of GSDMD in different treatment groups was detected by immunofluorescence technology,and the expression of IL-1β and IL-18 in each group was detected by immunohistochemical staining.Results:1.A subcutaneous tumor model of hypopharyngeal carcinoma in nude mice was successfully established by subcutaneous injection of Fadu cells into the back of the nude mice.2.The scanning electron microscope observation results showed that compared with 0.9% NS group,a certain amount of damage could be seen on the surface of cell membrane in all other treatment groups.Among which,EGFR-mAb/Au+NIR group had significantly more holes on the surface of cell membranes,and cell membranes were fragmented and bulbous,and a large amount of cell contents were released into the extracellular space,and the pseudopodium was largely reduced,degraded and broken.3.The results of immunofluorescence detection showed that in the direct injection treatment group The mean fluorescence intensity(MFI)of GSDMD in EGFR-mAb/Au.it+NIR was higher than that in other treatment groups,but there was no significant difference compared with EGFR-mAb/Au.it,AuNRs.it+NIR(P > 0.05).In the tail vein injection treatment group,the MFI of GSDMD in EGFR-mAb/Au.iv+NIR was significantly higher than that in other treatment groups(P < 0.0001).The results of different injection methods of the same drug showed that the MFI of GSDMD in the tumor body direct injection group was higher than that in the tail vein injection group(P < 0.05).4.Immunohistochemical staining results showed that for IL-1β,the mean optical density(MOD)of EGFR-mAb/Au.it+NIR was significantly higher than that of other treatment groups(P < 0.0001).In the tail vein injection treatment group,the MOD value of EGFR-mAb/Au.iv+NIR was higher than that of other treatment groups(P <0.05),but there was no significant difference compared with AuNRs.iv+NIR and EGFR-mAb/Au.iv+NIR(P > 0.05).Immunohistochemical staining results showed that for IL-18,the MOD value of EGFR-mAb/Au+NIR under two different injection modes was significantly higher than that of other treatment groups(P < 0.0001).The analysis of the results of different injection methods of the same drug showed that the MOD values of IL-1β and IL-18 in the tumor body direct injection group of EGFR-mAb/Au+NIR were higher than those in the tail vein injection group.Conclusion(s):1.EGFR-mAb/Au+NIR irradiation treatment can promote the destruction of Fadu cell membrane integrity,and then induce cell pyroptosis.2.Targeted photothermal effect of EGFR-mAb/AuNRs can induce the occurrence of pyrosis in hypopharyngeal squamous cell carcinoma cells in nude mice,promoting increased expression levels of pyrolytic protein GSDMD and inflammatory factor IL-1β and IL-18.3.The treatment effect of EGFR-mAb/Au+NIR by direct injection of tumor body for the treatment of hypopharyngeal cancer in nude mice is better than that by tail vein injection.4.These results indicate that EGFR-mAb/AuNRs targeted photothermal action is expected to be a novel approach for the treatment of hypopharyngeal cancer,providing a new decision for clinicians.