Construction For Prognostic Assessment Of Triple-negative Breast Cancer,Expressions And Clinical Significance Of Immune Checkpoint-related Proteins

PART1 Predictive Value of SII and CRP in Prognosis of Triple-negative Breast CancerObjective:Analyze the prognostic value of inflammatory parameters in the peripheral blood of patients with primary TNBC before surgery,and build a risk assessment model to explore convenient and effective clinical prognostic indicators,and provide a theoretical basis for timely predicting the risk of recurrence and metastasis of TNBC patients after surgery and improving the prognosis.Methods:The clinical data of 164 patients with primary TNBC treated from January2011 to September 2019 were retrospectively analyzed.Calculate the SII value,analyze the patient’s general data and peripheral blood parameters,and determine independent risk factors,the best clinical cut-off value and efficiencies of the variables in predicting the recurrence and metastasis were evaluated by receiver operating characteristic curve(ROC),Incorporate the independent risk factors established in Logistic regression into the Cox risk model to establish a risk prediction model.Results:1.Among the 164 patients with primary TNBC,48 had recurrence and metastasis within 5 years after surgery,accounting for 29.3%.Age ranges from 34 to 78 years old,median age is 54 years old,average age is(55.2±0.8)years old.SII value is(445.54±16.29)×10~9/L,CRP value is(2.0±0.1)mg/L.100 cases of menopause,64 cases of non-menopausal.120 cases of poorly differentiated tumors,44 cases of moderately differentiated,and 0 cases of highly differentiated.64 cases of lymph node metastases and 100 cases of non-metastatic tumors.16 cases of T1,124 cases of T2 and 24 cases of T3 stage.100 cases of N0,32 cases of N1,20 cases of N2,12 cases of N3 stage.16 cases ofⅠ,104 cases ofⅡ,44 cases ofⅢstage.132 cases of Ki-67>30%,32 cases of≤30%.2.TNM stage(OR=17.722,95%CI=2.587～121.392,P<0.05),SII(OR=1.005,95%CI=1.002～1.008,P<0.05)and CRP(OR=2.137,95%CI=1.385～3.298,P<0.05)were independent risk factors for postoperative recurrence and metastasis in TNBC patients by Logistic regression.3.SII predicts the prognosis AUC of 0.783(95%CI:0.706～0.859,P<0.05),the corresponding best cut-off value is 447.08,the sensitivity is 0.765,the specificity is0.752;The AUC of CRP is 0.753(95%CI:0.673～0.833,P<0.05),the corresponding best cut-off value is 1.95,the sensitivity is 0.804,the specificity is0.611.The AUC of SII-CRP is 0.832(95%CI:0.764～0.899,P<0.05),the corresponding best cut-off value is 0.27,the sensitivity is 0.843,the specificity is 0.726.4.Risk scoring model was created by incorporating TNM,SII and CRP into the Cox regression model,according to the HR values,a code of 1 was assigned to CRP>1.95(HR=2.297,95%CI=1.113～4.739,P<0.05),2 was assigned to SII>447.08(HR=5.490,95%CI=2.674～11.273,P<0.05),and 3 was assigned to TNMⅢstage(HR=15.546,95%CI=7.537～32.063,P<0.05),otherwise 0 was assigned.According to this model,the 1,3 and 5-year DFS rates were 77.1%、44.0%、32.4%in patients in high risk group(score≥3),and were 100.0%、96.6%、94.0%in patients in low risk group(score<3),the mean DFS were 44.2 and 97.8 months,respectively.The difference had statistical significance(P<0.05).Conclusion:1.TNM stage,preoperative SII value and preoperative CRP value are independent risk factors for postoperative recurrence and metastasis of primary triple-negative breast cancer.2.The predictive value of preoperative SII value combined with preoperative CRP value is better than that of a single index.3.The constructed risk model can stratify the prognosis of patients with triple-negative breast cancer,and provide help for clinical diagnosis and treatment and predicting the risk of recurrence and metastasis.PART2 Expression and Clinical Significance of LAG-3,PD-1/PD-L1 and TIL in Triple-negative Breast CancerObjective:To investigate the expressions of LAG-3,PD-1/PD-L1 and TIL in triple negative breast cancer(TNBC)and their clinical significance.Methods:Immunohistochemical method was used to detect the expression of LAG-3,PD-1 and PD-L1 in 62 cases of primary TNBC tissues and 30 cases of adjacent tissues.Hematoxylin-eosin method was used to evaluate TIL expression levels.χ~2 method was used to analyze the relationship between LAG-3 and various clinical factors and pathological characteristics.The influence of the four and their co-expression on the prognosis of TNBC was analyzed by single factor and Cox multivariate method,and the survival curve was drawn by the Kaplan-Meier method.Results:1.The positive expression rates of LAG-3 in TNBC and adjacent tissues are respectively 21.0%(13/62)and 0,P=0.008.The positive expression rates of PD-1 in TNBC and adjacent tissues are respectively 32.3%(20/62)and 0,P<0.001.The positive expression rates of PD-L1 in TNBC and adjacent tissues are respectively 25.8%(16/62)and 0,P=0.001.2.LAG-3 expression is related to lymph node metastasis(χ~2=4.219,P=0.040),Ki-67(χ~2=6.076,P=0.014),PD-1(χ~2=8.260,P=0.004),PD-L1(χ~2=8.735,P=0.003),TIL expressions(χ~2=8.269,P=0.004)in TNBC patients.3.Univariate analysis showed that DFS in TNBC patients was related to lymph node metastasis(χ~2=6.587,P=0.010),TNM stage(χ~2=16.291,P<0.001),Ki-67(χ~2=5.993,P=0.014),TIL(χ~2=17.040,P < 0.001),LAG-3/PD-1(χ~2=10.794,P=0.001),LAG-3/PD-L1(χ~2=13.612,P<0.001),LAG-3/TIL(χ~2=9.018,P=0.003).4.Multivariate analysis showed that TNM stage(HR=4.523,95%CI=1.571～13.022,P=0.005),LAG-3/PD-L1(HR=3.769,95%CI=1.368～10.386,P=0.010)and TIL(HR=4.389,95%CI=1.391～13.848,P=0.012)are independent risk factors for the prognosis of TNBC patients.Conclusion:1.The expressions of LAG-3,PD-1 and PD-L1 in TNBC tissue were significantly higher than those in adjacent tissues.2.LAG-3 expression is related to lymph node metastasis,Ki-67,PD-1,PD-L1,TIL expressions in TNBC patients.3.The prognosis of TNBC patients was related to lymph node metastasis,TNM stage,Ki-67,TIL,LAG-3/PD-1,LAG-3/PD-L1,LAG-3/TIL.4.TNM stage,LAG-3/PD-L1 and TIL are independent risk factors for the prognosis of TNBC patients,Although LAG-3 is not an independent risk factor,the co-expression of LAG-3 combined with PD-L1 is an independent factor.High TNM stage,LAG-3(+)PD-L1(+)and low TIL expression are related to the poor prognosis of triple-negative breast cancer patients.5.The combined co-expression of LAG-3 and PD-L1 can provide more theoretical basis for the clinical application of immune checkpoint double block in patients with triple-negative breast cancer.Anti-LAG-3 and anti-PD-1/PD-L1 dual blockade may become a breakthrough in triple-negative breast cancer immunotherapy.