Expression And Clinical Significance Of FOXP3 And BTLA In Esophageal Squamous Cell Carcinoma

Objective:Esophageal cancer is a common gastrointestinal cancer.China is a high incidence area.The vast majority is squamous cell carcinoma.The occurrence, development,invasion and metastasis of esophageal cancer is a multi-gene,multi-step process of working together,but the specific mechanism is not yet clear.In recent years scholars have noted that immunity played an important role for the diagnosis and treatment of esophageal cancer.The role in tumor immunity of FOXP3 gene found in 2001 and BTLA gene found in 2003 is increasing more and more people's attention.They play immunosuppressive effects by different but interrelated mechanisms.Our purpose is to establish a real-time quantitative reverse transcription-polymerase chain reaction(RT-PCR) method for detecting the expression of FOXP3(forkhead box P3) gene and BTLA(B and T lymphocyte attenuator,BTLA ) gene in Esophageal squamous cell carcinoma and explore the relationship between mRNA expression and the occurrence,development of esophagus squamous carcinoma to provide a new direction for tumor immunotherapy of esophageal cancer.Methods:We have collected 42 cases of surgical resection of esophageal cancer samples and 30 samples of control organizations.Based on fluorescent TaqMan methodology,a real-time quantitative RT-PCR was set up.In this method,a cloning vector pMD 18-T-FOXP3 and pMD 18-T-BTLA were constructed as standard plasmids.The specific expression of FOXP3 and BTLA in 42 patients with ESSA and 30 healthy controls were measured by using GeneAmp 7000 Sequence Detection Systems.We analyzed the relationship between expression of FOXP3 and BTLA in 42 cases and clinical pathological information of esophageal cancer.All statistical analyses were conducted by SPSS for windows 11.0 statistical software system.That measurement data used 11 Comparison between the two groups using t-test.The relationships between FOXP3 and BTLA's expression were analysed by using linear statistics.The results are P＜0.05 for the difference. Results:The FOXP3 mRNA copy number ranged from 3.4×10~4～5.3×10~6copy/μg RNA in 42 esophageal cancer patients and 5.6×102～1.2×104copy/μg RNA in 30 healthy controls.The mean FOXP3 mRNA copy number in esophageal cancer group was significantly higher than that in controls[(7.2±2.6)×10~5vs(6.8±3.4)×10~3; P＜0.01].At the same time,the BTLA mRNA copy number ranged from 5.8×10~3～2.7×10~5 copy/μg RNA in 42 esophageal cancer patients and 2.5×10~2～6.1×10~4copy/μg RNA in 30 healthy controls.Also,The mean BTLA mRNA copy number in esophageal cancer group was significantly higher than that in controls[(8.8±2.6)×10~4vs(9.2±3.4)×10~3;P＜0.01].There were no significant differences in the mean FOXP3 mRNA and BTLA mRNA copy number between the groups of age,sex,tumor size and level of differentiation lipids(P＞0.05).But dependablity was found in TNM staging and region lymph node metastasis significantly(P＜0.01).Conclusion:1.A real-time quantitative RT-PCR method for detecting the expression of FOXP3 gene and BTLA gene have been successfully established.2.The mean FOXP3 mRNA copy number in esophageal cancer group was significantly higher than that in controls.3.There were no significant differences in the mean FOXP3 mRNA copy number between the groups of age,sex,tumor size and level of differentiation lipids.But dependablity was found in TNM staging and region lymph node metastasis significantly.The mean FOXP3 mRNA copy number in the groups of region lymph node metastasis or TNM stagingⅡB-Ⅲwas significantly higher than that in the groups of non-region lymph node metastasis or TNM stagingⅠ-ⅡA.4.The mean BTLA mRNA copy number in esophageal cancer group was significantly higher than that in controls.5.There were no significant differences in the mean BTLA mRNA copy number between the groups of age,sex,tumor size and level of differentiation lipids.But dependablity was found in TNM staging and region lymph node metastasis significantly.The mean BTLA mRNA copy number in the groups of region lymph node metastasis or TNM stagingⅡB-Ⅲwas significantly higher than that in the groups of non-region lymph node metastasis or TNM stagingⅠ-ⅡA.6.The mean FOXP3 and BTLA mRNA copy number in esophageal cancer have a linear relationship.But the correlation is not strong since there are some other factors between the interference.