| Objective:Endometrial carcinoma（EC）is a common malignant tumor of the female reproductive system.Domestic and foreign research data show that the morbidity and mortality of EC continue to rise,seriously endangering the health of women.Our previous experiments found that visfatin can regulate the proliferation and apoptosis of EC cells through PI3K/MAPK pathway,and it has a correlation with the infiltration of CD4+T cells.Helper T cell 9（Th9）,a new type of CD4+T cell subset,with PU.1 as its specific transcription,can affect the tumor cells directly or indirectly influence the proliferation and apoptosis of tumor cells by regulating innate immune response and adaptive immune response,but the specific role of Th9 cells in EC has not been clarified.Therefore,we detect the expression of visfatin,Th9 cells as well as PU.1 in peripheral blood（PB）before operation and expression of Th9 cells and PU.1in endometrium tissue from normal controls（NC）,patients with atypical hyperplasia（AH）and EC in operation in order to explore whether there are differences in the level of visfatin,the proportion of Th9 cells and the expression of PU.1 between EC patients and the other two groups,whether there is a correlation between visfatin and Th9 cells,and the relationship between them and the clinicopathological features of EC patients.This research is helpful to improve the understanding of the mechanism of EC progression and provide a theoretical basis for EC immunotherapy.Methods:The PB samples before operation,endometrium tissue in operation and related case data of 32 patients with EC and 13 patients with AH treated in the Department of Gynecology and Obstetrics of General Hospital of Tianjin Medical University from January 28,2019 to January 31,2020,as well as and 12 matched NC objectives were collected.ELISA was used to detect the level of visfatin in PB.Mass spectrometry flow cytometry（Cy TOF）was used to detect the proportion of Th9 cells.The m RNA expression levels of PU.1,key transcription factor of Th9 cells,were also detected by quantitative real-time reverse transcription polymerase chain reaction（q RT-PCR）.All the experimental data are regarded as measurement data and expressed by mean±standard deviation.Graph Pad Prism 6 software was used for statistical analysis,and the differences among the three groups and between the two groups were tested by ANOVA analysis and independent sample t-test,respectively.Spearman correlation coefficient was used to evaluate the correlation between visfatin and Th9 cells to explore whether visfatin and Th9 cells complement or antagonize each other in the occurrence and development of EC.Results:1.The results of ELISA showed that the level of visfatin in PB of NC group（6.92±0.48 ng/ml）,AH group（10.26±0.28 ng/ml）and EC group（20.53±0.68 ng/ml）was gradually increasing,and there was significant statistical difference（P<0.05）.2.The results of Cy TOF flow showed that the proportion of CD4+T cells in PB and endometrium tissue of NC group,AH group and EC group was gradually decreasing without significant statistical difference（P>0.05）.The proportion of Th9 cells in PB of NC group（5.65±0.63%）,AH group（3.29±0.53%）and EC group（2.43±0.44%）was also gradually decreasing with significant statistical difference（P<0.05）.Moreover,the proportion of Th9 cells in PB was significantly different in EC patients with different histological grades（P<0.05）.The proportion of Th9 cells in PB of EC patients with the histological grade of G2（2.60±0.56%）was significantly lower than that of G1（2.49±0.99%）with P value lower than 0.05.The proportion of Th9 cells in the intimal tissue also showed a gradually decreasing trend in the NC group（9.01±0.98%）,AH group（7.38±1.73%）and EC group（4.43±0.85%）,and the difference was statistically significant（P<0.05）.The results of q RT-PCR detection showed that the expression level of PU.1 in PB（0.137±0.015）and endometrium tissue（0.340±0.057）of patients from EC group was lower than PB（1.002±0.022）and endometrium tissue（1.004±0.018）of NC group with statistically significant difference（P<0.05）.3.The results of correlation analysis showed The expression level of visfatin in PB was negatively correlated with the proportion of Th9 cells in the overall study subjects（r=-0.800,P<0.05）.However,no association was found between the two indices in NC group,AH group and EC group with r NC=-0.619,r AH=0.15,and r EC=-0.064.And there was no statistical difference.Conclusion:1.The proportion of Th9 cells in PB and endometrium tissue from NC,AH,and EC group showed a decreasing trend,suggesting that Th9 cells may play an anti-tumor role in the carcinogenesis of EC.In addition,proportion of Th9 cells in PB has some implications for the clinicopathological characteristics of EC patients.2.The expression level of transcription factor PU.1 was significantly lower in PB and tissues of patients with EC than those in NC group,which is consistent with the trend of Th9 cells.This further confirmed the decrease in the number of Th9 cells and speculated that its function might also be restricted.3.Visfatin played a promoting role in the occurrence of EC.There was a negative correlation between the expression of visfatin and the proportion of Th9 cells in the PB of overall objectives indicating that there might was a relationship and interaction between them in the occurrence and process of EC. |
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