Study On The Expression Of Trop-2 In Differentiated Oral Squamous Cell Carcinoma And Efficacy Of A Novel Adenovirus With Human IL-13 In Oral Cancer

Research 1:Correlation between the expression of Trop-2 in differented oral squamous cell carcinoma and its mechanical stiffnessBackground and Objective: Human trophoblast cell surface antigen 2(Trop-2)is highly expressed in many squamous cell carcinomas and plays an important role in promoting tumor occurrence,development and invasion,and is closely related to the survival rate of patients.However,there are few studies on the relationship between the expression of Trop-2 and the occurrence and development of in oral squamous cell carcinoma(OSCC).At present,most of the research on the diagnosis of OSCC are mainly based on the molecular biology level,focusing on the study and discovery of cancer-specific markers.However,more and more studies have shown that the mechanical properties of tissue interface are closely involved in the biological process of proliferation,differentiation,movement and migration of cancer cells.Therefore,it is necessary to explore the correlation between the expression of specific molecular marker Trop-2 and the interfacial mechanical properties of OSCC with differented degrees of differentiation.Based on this,the purpose of this study was to observe the relationship between the expression of specific marker Trop-2 and the changes of mechanical stiffness of OSCC.In order to explore the biomechanical coupling mechanism between the changes of mechanical properties and the participation of specific target molecules in the carcinogenesis and development of OSCC,which provides a new perspective and useful supplement for the early accurate diagnosis and curative effect evaluation of OSCC.Material and Methods: A total of 108 OSCC specimens from the Second Affiliated Hospital of Lanzhou University from January 2012 to March 2018 were selected as experimental groups.HE staining was used to determine the degree of OSCC differentiation,and immunohistochemical staining was used to detect different differentiation.The extent of expression of Trop-2 protein in OSCC tissues,Kaplan-Meier calculation of patient survival,analysis of the relationship between Trop-2 expression and clinicopathological features of OSCC,evaluation of the effect of Trop-2 expression on tumor prognosis;by atomic force microscopy(AFM)The micro-nano morphology and tissue stiffness of OSCC tissue samples with different degrees of differentiation were observed.Results: In different degrees of OSCC tissues,the expression of Trop-2 was significantly increased in poorly differentiated oral squamous cell carcinoma.Kaplan-Meier curves showed that with the increase of Trop-2 expression,the overall survival rate of patients decreased gradually.There was a negative correlation.AFM results showed that there were significant differences in the mechanical stiffness of OSCC with different degrees of differentiation.The OSCC of the poorly differentiated OSCC had higher stiffness and the stiffness of the OSCC was smaller,that is,the poorly differentiated OSCC tissue was more highly differentiated than the OSCC tissue."Soft".Pearson correlation analysis showed that there was a negative correlation between the expression of Trop-2 and the interfacial stiffness of OSCC tissues with different degrees of OSCC tissues.Conclusion: High expression of Trop-2 in oral squamous cell carcinoma is closely related to patient survival.There was a significant negative correlation between the changes in tissue mechanical stiffness and the difference in Trop-2 expression in OSCC with low,medium and high differentiation.Therefore,we speculate that the specific marker Trop-2 expression in OSCC significantly affects the prognosis of patients,and can be used as a potential target for the treatment of oral squamous cell carcinoma;the expression of Trop-2 can regulate the mechanical stiffness of cancer tissue interface and participate in oral squamous cell carcinoma.The process of development,invasion and transfer occurs.Compared with traditional molecular biology research,this provides a new perspective for accurate diagnosis and evaluation of oral squamous cell carcinoma.Research 2 :A new oncolytic adenovirus armed with IL-13 could improve the efficacy in oral cancer modelBackground and Objective:Oral carcinoma(OC)is the most common cancer of squamous cell carcinoma.The traditional therapies effect is always limited.Oncolytic virus is a novel strategy for cancer therapy and show potential to use in OC treatment.Even the oncolytic adenovirus has been studied for a long time,but clinical application of adenovirus was limited by pre-existing neutralizing antibodies.So the purpose of this study was to test a new modified adenovirus potency in OC in vitro and in vivo.Material and Methods: A multiple modified adenovirus(MMAD)armed with IL-13 was constructed and tested function in Cal-27.The potency of MMAD-IL-13 were tested in vitro and in vivo.In vitro experiment used CCK-8 kit to detect the IC50 in varies OC cell lines.Cal-27 xenograft model was used to determine the anti-tumor effect of MMAd-IL-13 in vivo.Apoptosis was measured in Cal-27 by WB assay.Immunity response was detected in Cal-27 xenograft model received intratumor injection after 7 d.The potency of MMAD and MMAD-IL-13 were further compared in Cal-27 PBMC model.Results: MMAD-IL-13 was successfully constructed,the virus harvested could replicated and overexpress human IL-13 in Cal-27.Armed with IL-13 was able to enhance the anti-tumor effect in vitro by induce apoptosis and reduced M2 percent in tumor environment in vivo.The MMAD-IL-13 also showed great anti-tumor effect in Cal-27,SCC-4,Tca8113 in vitro and Cal-27 xenograft model in vivo.Meanwhile,MMAD-IL13 didn`t harm normal human oral epithelial cell in vitro and it was no effect on body weight in Cal-27 xenograft model.In Cal-27 PBMC model,MMAD-IL-13 show stronger anti-tumor effect than MMAD.Conclusion: A new oncolytic adenovirus carry the human IL-13 gene was constructed.This virus can effective remission the tumor development and kill the cancer cell in vivo and in vitro,has the potential for clinical therapy.