Expression And Significance Of Immune Checkpoint TIM-3,LAG-3,TIGIT,VISTA And PD-1 In Cervical Squamous Cell Carcinoma

Objective:Analysis of the difference in expression of T cell immunoglobulin mucin 3(TIM-3)、lymphocyte activation gene 3(LAG-3),T-cell immunoglobulin and immune receptor tyrosine-based inhibitory motif domain(TIGIT),V-domain Ig suppressor of T-cell activation(VISTA)and Programmed Death receptor-1(PD-1)in normal cervix,cervical intraepithelial neoplasia and cervical squamous cell carcinoma,To explore its role in the occurrence and development of cervical cancer and its influence on the prognosis of cervical cancer,and provide theoretical basis for combined immunotherapy of cervical squamous cell carcinoma.Methods:Ninety paraffin specimens of Cervical tissue were collected,including 40 patients with Cervical squamous cell carcinoma who underwent surgical treatment in the Department of Gynecology,Affiliated Hospital of Qingdao University from February2013 to September 2016 were selected as the experimental group,and 20 patients with Cervical intraepithelial neoplasia(CIN)were selected.Postoperative pathology confirmed grade CINⅡ-Ⅲ and 30 normal cervical tissues negative for human papilloma virus(HPV)infection as controls.Immunohistochemical staining was used to detect the expression of TIM-3,LAG-3,TIGIT,VISTA and PD-1 in 40 cases of cervical neoplasm,20 cases of cervical squamous epithelial lesion and 30 cases of normal cervical tissue.Statistical indicators were analyzed according to the clinicopathological characteristics of the patients.The patients were followed up closely for a long time,and the long-term prognosis was statistically evaluated.Results:1.There were significant differences in immune checkpoint between the experimental group and the control group.The positive rates of TIM-3,LAG-3,TIGIT,VISTA and PD-1 in cervical squamous cell carcinoma tissues were 57.5%(23/40),65%(26/40),60%(24/40),60%(24/40)和65%(26/40)respectively;The expression rates in CIN group were 25%(5/20),30%(6/20),25%(5/20),25%(5/20)和25%(5/20)respectively;The positive expression rates in normal cervical tissues were3.3%(1/30),3.3%(1/30),3.3%(1/30),3.3%(1/30)and 3.3%(1/30).The positive expression rate of immune checkpoint in cervical squamous cell carcinoma was higher than CIN group and normal group,CIN group was higher than normal cervical group,the difference was statistically significant(P<0.05).2.In the clinicopathological analysis of 40 cases of cervical squamous cell carcinoma,the significant positive expression of TIM-3 was mainly related to the depth of tumor cell invasion(P < 0.05),and the positive expression of TIGIT was also related to the depth of tumor invasion(P < 0.05).However,the high expression of TIM-3 and TIGIT may not be associated with clinicopathological characteristics such as patient age,tumor diameter,FIGO stage,differentiation degree,lymph node metastasis and vascular invasion depth(ALL P >0.05).The positive expression of LAG-3 、VISTA and PD-1was not associated with clinicopathological factors(ALL P >0.05).3.The median follow-up time of 40 patients with cervical squamous cell carcinoma was 30 months(1-62 months).Kaplan-meier survival results showed that TIM-3,LAG-3,VISTA and PD-1 of Progression free survival with negative expression were significantly longer than those with positive expression,and the difference was statistically significant,the difference was statistically significant(P<0.05),and TIGIT positive expression was not associated with survival prognosis(P>0.05).Single factor analysis showed that TIM-3,VISTA and PD-1 proteins were positively expressed,and the survival time of patients with lymph node metastasis and poorly differentiated cervical squamous cell carcinoma was significantly shorter than that of patients with negative expression of TIM-3,VISTA and PD-1 proteins,no lymph node metastasis,and medium and high differentiation degree.Multivariate regression showed that TIM-3 positive and lymph node metastasis were independent potential risk factors for long-term survival of patients with cervical squamous cell carcinoma(P < 0.05).Spearman rank correlation regression analysis showed that there was a positive linear correlation between VISTA and PD-1 immune protein expression(r = 0.364,P < 0.05),and the difference was statistically significant.Conclusion:1.The positive expression rates of TIM-3,LAG-3,TIGIT,VISTA and PD-1 in cervical squamous cell carcinoma were higher than CIN group and normal group,and CIN group was higher than normal cervical group,the differences were statistically significant,suggesting that these five immune regulatory proteins may be related to the occurrence and development of cervical squamous cell carcinoma and play a role in the immune escape of cervical lesions.It provides experimental basis for immunotherapy of cervical cancer and is a potential immunotherapy target.2 The expressions of TIM-3 and TIGIT were correlated with clinicopathological features,and TIM-3,LAG-3,VISTA and PD-1 were correlated with PFS,with statistically significant differences.3 There was a positive correlation between VISTA and PD-1 expression in cervical squamous cell carcinoma,and the difference was statistically significant.