The Detection Of T Lymphocyte Phenotype In Peripheral Blood Of Oral Carcinoma Patients Before And After Treatment

Objective: Oral and maxillofacial cancinomas are common malignant tumors in our country,such as tongue cancer,gum cancer,buccal cancer, and so on,but its incidence is caused by a variety of factors,such as environmental factors,psychological factors,immune factors and so on. Many scholars pay more attention to the immune factors. immune function of Patients with oral and maxillofacial malignant tumors is likely to be lower than the normal crowd in early or advanced period,particularly in advanced period. Therefore,the body's immune state in the process of occurrence and development of malignant tumors have a certain reference value. Fas - FasL system is an important way of apoptosis in our body. Moreover,apoptosis control is an important factor in the development of tumor,so Fas - FasL apoptosis has become a hot point in medical research in recent years,the abnormal expression of Fas and its ligand FasL system is involved not only in the process of abnormal regulation of the apoptosis of tumor,but also in tumor immune mechanisms escape from the kill of the immune system. The expression of Fas and FasL is an important part in the body between the struggle of immune system in our body and apoptosis systems of the tumor,this test is aimed at detecting the expression of the T-lymphocyte subtypes in the peripheral blood of patients with oral cancer by flow cytometry before and after the treatment,particularly at detecting the expression of CD4, Fas and FasL in the peripheral blood of patients with oral cancer,and at exploring the role of Fas and FasL in the pathogenesis of oral cancer patients,so that the pathogenesis of oral cancer may get further development from the point of view in immunology and provide comprehensive treatment of oral cancer with a theoretical basis. Methods: Fresh blood pleasing 100μl, add FITC-CD3 monoclonal antibody 10μl, FITC labeled CD4 monoclonal antibody 10μl, PE labeled CD8 monoclonal antibody 10μl and PE labeled CD56 monoclonal antibody 10μl, PE labeled Fas monoclonal antibody 10μl, FITC labeled CD4 monoclonal antibody and PE labeled Fas monoclonal antibody 10μl seprately, PE labeled FasL monoclonal antibody 10μl, FITC labeled CD4 monoclonal antibody and PE labeled FasL monoclonal antibody 10μl seprately, incubated at room temperature for one dark hour, to add red blood cells with lysis buffer, lysis of RBC 5 minutes, add PBS solution hemoglobin washed and re-suspended in Medium 1ml PBS to flow cytometry T lymphocyte phenotype accounted for the percentage of total lymphocytes and T lymphocyte cells of Fas+ and FasL+ T lymphocytes and the percentage of CD4+/Fas+ and CD4+/FasL+T lymphoid the proportion of cells and calculate the CD4+T cells/CD8 + T cells ratio.Results: 1 An oral cancer patients'changes in immune parameters before and after the treatment(invoved both surgery and chemotherapy). 1.1 The CD3 +,CD4 +,CD8 +,CD56 + T lymphocyte percentage in peripheral blood of oral carcinoma patients in a week after surgery were 63.23±4.96%,42.37±3.96%,24.46±3.86%,14.60±5.17%,CD4 + / CD8 + ratio ranged from1.77±0.34; the expression of CD3 compared with the preoperative data showed that although there was small-scale improvement in peripheral blood of oral carcinoma patients, but by the statistical analysis the difference between before surgery and after surgery group there were no significant difference(P﹥0.05 ),while the expression of CD3 was still lower than the control group (P﹤0.05) . The expression of CD4 in a week after surgery in peripheral blood of oral carcinoma patients was still lower than the normal control group, but compared with the preoperative data it was upgrade larger (P﹤0.05) , the expression of CD8 in peripheral blood of oral carcinoma patients was significantly lower than the preoperative group(P﹤0.05), but compared with the normal control group the expression of CD8 had no significant difference, CD4 / CD8 ratio also has been picked up than before surgery group, although its value has not yet to be resumed to the level of nomal control group (P <0.05),the expression of CD56 in peripheral blood of oral carcinoma patients was significantly lower than the state of the preoperative group(P﹤0.05),but compared with the normal control group it had no significant difference.1.2 oral cancer patients received chemotherapy by tegafur and oxaliplatin two weeks after surgery,then collected peripheral blood of the patients again after three weeks after chemotherapy, The CD3 +,CD4 +,CD8 +,CD56 + T lymphocyte percentage in peripheral blood of oral carcinoma patients after chemotherapy were 63.56±9.68%,37.55±9.13%,25.79±6.74%,18.61±8.23 %,CD4 + / CD8 + ratio ranged from 1.57±0.43; the expression of CD3 was similar to the patients of pre-chemotherapy group (P >0.05),the expression of CD4 is higher than the pre-chemotherapy group(P <0.05),CD8 expression in peripheral blood of oral carcinoma patients after chemotherapy was significantly decreased compared with pre-chemotherapy group (P <0.05),CD4 + / CD8 + ratio was significantly higher than before chemotherapy group (P <0.05),but the expression of CD56 had no significant change compared with before chemotherapy group.2 Oral cancer patients'Fas and FasL changes in peripheral blood before and after treatment(invoved both surgery and chemotherapy).2.1The CD4+/Fas+,Fas+,CD4+/FasL+,FasL+ T lymphocyte percentage in peripheral blood of oral carcinoma patients after surgery were 6.30±1.67%,1.98±0.93%,0.70±0.28%,0.99±0.28%,the expression of CD4+/Fas+ in peripheral blood of oral carcinoma patients was significantly lower than the preoperative state(P﹤0.05),but the expression of Fas+ in peripheral blood of oral carcinoma patients had no significant difference after surgery compared with the preoperative data (P >0.05 ); the expression of CD4+/FasL+ and FasL+ in peripheral blood of oral carcinoma patients were significantly increased compared with the preoperative data(P﹤0.05).2.2 oral cancer patients received chemotherapy by tegafur and oxaliplatin in two weeks after surgery,then collected the peripheral blood of the patients again after three weeks after chemotherapy, CD4+/Fas+,Fas+,CD4+/FasL+, FasL+ T lymphocyte percentage in peripheral blood of oral carcinoma patients after chemotherapy were 8.36±2.12% , 2.01±1.19%, 0.62±0.19%, 0.95±0.23%,the expression of the CD4+/Fas+ and Fas+ in peripheral blood of oral carcinoma patients were significantly higher than before chemotherapy group(P﹤0.05),but the expression of the CD4+/FasL+ and FasL+ in peripheral blood of oral carcinoma patients after chemotherapy had no significant change compared with before chemotherapy group(P﹥0.05).3 The relationship among different clinical factors and the expression of Fas and FasL.3.1 The expression of CD4+/Fas+ and CD4+/FasL+ in peripheral blood of oral carcinoma patients was significantly higher than the normal control group(P﹤0.05),the expression of CD4+/Fas+ and CD4+/FasL+ in theⅢandⅣstage in peripheral blood of oral carcinoma patients was significantly higher thanⅠandⅡstage group(P﹤0.05); the expression Fas in peripheral blood of oral carcinoma patients with lymph node metastasis was significantly higher than patients with no lymph node metastasis group (P <0.05),but the expression of CD4+/FasL+ in peripheral blood of oral carcinoma patients with the lymph node metastasis had no significant difference compared with no lymph node metastasis group. the expression of CD4+/Fas+ in the tumor's diameterin larger than three centimeter group was higher than in the tumor's diameterin not larger than three centimeter group(P﹤0.05),while the expression of CD4+/FasL+ had no significant change in both of them(P﹤0.05).3.2 The different age,sex,smoking factors had no significant relation with the expression of Fas and FasL in peripheral blood of patients with oral cancer.Conclusion: 1 There was a different degree immune suppression in patients with oral carcinoma compared with normal control group. Supporting T cells reduced, and suppressor T-cell increased,surgery and chemotherapy may relieve some of immune suppression. so surgery and chemotherapy was effective for the treatment of patients with oral cancer.2 The expression of Fas in oral cancer patients'peripheral blood may have relationship with the tumor stage, whether there was lymph node metastasis and tumor size,perhaps the expression of Fas could be made as an immunological marker of oral carcinoma diagnosis.3 Fas / FasL system is a double-edged sword in our body, the immune cells can remove tumor cells of the body, while tumor cells can also remove the activated T lymphocytes by the high expression of FasL in peripheral blood of oral carcinoma patients,but the pathogenesis of it was very complex and need further study.