| Background and objectiveAfter the human body is stimulated by internal or external factors for a long time,the carcinogen in the tissue cells will be activated,and then through a series of complex and interleaved pathways and mechanisms,the tissue cells will proliferate abnormally,and the body's systemic immune function will be accompanied.Eventually,tumor occurs.Under normal physiological conditions,lymphocytes and other immune cells in the body work together to maintain the normal operation of the body's immune function,which can resist the invasion of foreign antigens and prevent the occurrence of autoimmune diseases.T cells are the main components of the body's immune cells.Long-term exposure to tumor antigens can cause dysfunction of the T cells,and their effector functions can be inhibited,allowing the tumor cells and tissues to escape from the surveillance of the body's immune system and further develop.According to the 2018 National Epidemiological Survey,around 830,000people worldwide suffer from head and neck cancer each year,and the number of deaths due to head and neck cancer is as high as 430,000 in the same year.Head neck squamous cell carcinoma accounts for about 95%of the total number of head and neck cancer cases,of which oral squamous cell carcinoma(OSCC)is the most common type of disease in HNSCC.Its incidence rate has increased gradually in recent years.At present,the main treatment method is surgical treatment,which is supplemented by radiotherapy and chemotherapy in light of the location and malignancy of the tumor.But there is still a lack of effective targeted therapy,and the local and systemic factors of its outcome are complicated,so the survival rate and prognosis of OSCC are still not optimistic.With the development of immunology,researchers believe that the insignificant effect and poor prognosis of OSCC chemotherapy and targeted therapy may be related to the reason that the dysfunction of immune cells(especially T cells)in the systemic circulation and local microenvironment of the patient causes the suppression of immune function.Among them,PD-1,TIM-3,CTLA-4,etc.are common tumor negative costimulatory factors on T cells.There are many studies on the expression of PD-1 and TIM-3 in lung cancer,gastric cancer,small cell cancer and other tumors.And PD-1/PD-L1monoclonal antibody therapy has been used in clinical treatment of various tumors[2],such as breast ductal carcinoma,non-small cell carcinoma and so on.However,the expression status of PD-1 and TIM-3 in the field of oral squamous cell carcinoma,especially on T cells,is less studied.According to the previous research background,this paper intends to use flow cytometry to detect the expression of PD-1 and TIM-3on T cells in OSCC peripheral blood and cancer tissues of OSCC patients,analyze the relationship between the two immune checkpoint molecules and their correlation with different clinicopathological features of OSCC patients,and explore the relationship between local and systemic immune function and disease occurrence and progression in patients with oral squamous cell carcinoma,providing a theoretical basis for the future research and application of OSCC targeted therapy.Methods1.Chose untreated oral squamous cell carcinoma patients(41 cases)and healthy medical examiners(20 cases)from the First Affiliated Hospital of Zhengzhou University and the Department of Stomatology of Henan Provincial Stomatological Hospital,collected the peripheral blood specimens and recorded the characteristics of clinical cases corresponding to each patients.Used flow cytometry to detect the expression of PD-1 and TIM-3 on the peripheral blood T cells between the experimental group and the control group and applied SPSS to finish the data collection,collation and analysis.Studied and discussed the relationship between the the two on peripheral blood T cells and clinical pathological features such as TNM stage and pathological grade..2.Since some OSCC patients'lesions were small,only 25 of the 41 OSCC patients mentioned above were collected,and fresh tissues were collected during the corresponding operation too,namely cancer tissues(experimental group)and paracancerous tissues(control group).Used flow cytometry to detect the expression of PD-1 and TIM-3 on the peripheral blood T cells between the experimental group and the control group and applied SPSS to finish the data collection,collation and analysis.Studied and discussed the correlation between the two on peripheral blood T cells and clinical pathological features such as TNM stage and pathological grade.3.The data obtained in this experiment was analyzed by SPSS22.0 software.Quantitative data which passed the normality test was analyzed by t-test,and Spearman analysis method was used to correlation analysis.(?=0.05,P<0.05)Results1.The expression of PD-1 and TIM-3 on the peripheral blood T cells of oral squamous cell carcinoma group and healthy control group were significantly different.The expression rate of PD-1 on the peripheral blood CD4~+T cells of the OSCC group was 10.276±2.352%,higher than that of the healthy control group(3.740±0.862%);the expression rate of PD-1 on peripheral blood CD8~+T cells of oral squamous cell carcinoma group was 11.985±2.925%,higher than that of the healthy control group(4.050±0.954%);the expression rate of TIM-3 on the peripheral blood CD4~+T cells of the oral squamous cell carcinoma group was 4.024±0.680%,higher than that of the healthy control group(0.940±0.424%);the expression rate of TIM-3 on peripheral blood CD8~+T cells of oral squamous cell carcinoma group was 4.217±0.716%,higher than that of the healthy control group(1.240±0.652%).The differences were statistically significant.(P<0.05)2.The expression of PD-1 and TIM-3 on T cells in cancer tissues and adjacent tissues is significantly different.The expression rate of PD-1 on CD4~+T cells in cancer tissues was 30.284±5.292%,higher than that in adjacent tissues(10.680±4.731%);the expression rate of PD-1 on CD8~+T cells in cancer tissues was30.080±5.933%,higher than that in adjacent tissues(10.896±1.754%);the expression rate of TIM-3 on CD4~+T cells in cancer tissues was 23.032±3.606%,higher than that in adjacent tissues(10.844±2.034%);the expression rate of TIM-3 on CD8~+T cells in cancer tissues was 24.328±3.825%,higher than that in adjacent tissues(10.624±1.279%).The differences are statistically significant.(P<0.05)3.The expression of PD-1 on CD4~+T,CD8~+T in peripheral blood of OSCC patients was correlated with the patient's TNM classification and the presence or absence of lymph node metastasis.(P<0.05);the expression of TIM-3 on CD4~+T,CD8~+T in peripheral blood of OSCC patients was related to the presence or absence of lymph node metastasis.(P<0.05);the expression of PD-1 and TIM-3 on T cells in peripheral blood of OSCC patients was not significantly correlated with the patient's gender,age,primary tumor site,etc.(P>0.05);4.The expression of PD-1 on CD8~+T in cancer tissues of OSCC patients was related to the presence or absence of lymph node metastasis and TNM classification.(P<0.05);TIM-3 on T cells in cancer tissues was not significantly correlated with the patient'sto the presence or absence of lymph node metastasis or TNM classification,etc.(P>0.05)5.In peripheral blood of patients with oral squamous cell carcinoma,the expression of PD-1 on CD4~+T cells was positively correlated with TIM-3(r_s=0.461,P=0.002<0.05),and the expression of PD-1 on CD8~+T cells was positively correlated with TIM-3(r_s=0.337,P=0.031<0.05).6.In cancer tissues of patients,the expression of PD-1 on CD4~+T cells was positively correlated with TIM-3(r_s=0.388,P=0.012),and the expression of PD-1 on CD8~+T cells was positively correlated with TIM-3(r_s=0.401,P=0.003<0.05).Conclusions1.The expression levels of PD-1 and TIM-3 on the CD4~+T and CD8~+T cells in peripheral blood of OSCC patients were significantly higher than those in healthy control group,indicating that there may be immunosuppression in the systemic circulation of OSCC patients.2.The expression levels of PD-1 and TIM-3 on tne CD4~+T and CD8~+T cells of T in cancer tissues of OSCC patients were significantly higher than those in normal adjacent tissues,indicating that the local tumor microenvironment of patients with oral squamous cell carcinoma may show an immunosuppressive state,which promotes the tumor to escape from the immune surveillance and further develop.3.The expression of PD-1 and TIM-3 on T cells in peripheral blood and cancer tissues of OSCC patients was related to the presence or absence of lymph node metastasis and TNM stage of OSCC,indicating that the development of OSCC may be related to PD-1 and TIM-3.4.The expression of PD-1 on T cells in peripheral blood and cancer tissues of OSCC patients was positively correlated with the expression of TIM-3,indicating that PD-1 and TIM-3 are co-expressed in OSCC,and may have a synergistic effect on tumor progression.5.PD-1 and TIM-3 may provide new directions for the diagnosis,treatment and prognosis evaluation of OSCC. |
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