The Correlation Of Margin Status,Pattern Of Invasion And Tumor-associated Neutrophils And Their Influence On Prognosis Of Oral Squamous Cell Carcinoma

BackgroundThorough removal of rumor during initial surgical treatment is crucial for oral squamous cell carcinoma(OSCC)local control.To access the thoroughness of surgery,margin status,including margins at the epithelium and basement,is a valuable indicator.However,the criteria of surgical margins have not yet been uniformed.For epithelial margin,margins with medium-to-severe dysplasia and/or residual cancer are recognized prognostic factor OSCC,but the treatment of mild dysplasia margin is controversial.The management of margins at the invasion front is difficult during OSCC operation.At present margin more than 5mm from tumor is defined as negative margin.However,even for such negative margins good prognosis is not guaranteed.This may be related to the biological behavior of the tumor.The ability of tumor to infiltrate into deep normal tissue represents the invasiveness of tumor and is directly related to invasion pattern,which is also the result of interaction between tumor and tumor microenvironment.This study mainly focuses on margin status,worst pattern of invasion,and their prognostic value of OSCC,as well as the relation between infiltrating tumor-associated neutrophils(TANs)and WPOI.OneThe influence of margin status on the prognosis of oral squamous cell carcinoma[Objective]To evaluate the clinical relevance between different margin statuses,especially mild dysplasia,and prognosis in patients undergoing primary surgical treatment for oral squamous cell carcinoma(OSCC).[Methods]Five hundred and thirty-nine patients with T1-4NOMQ OSCC,who underwent surgery alone at a stomatology hospital in Nanjing,China during the years 2005—2014,were included.Tumor and margin status were assessed.Overall survival(OS),disease-free survival(DFS),and recurrence-free survival(RFS)were calculated by Kaplan-Meier method.Predictors of RFS,OS,and DFS were analyzed.[Results]Positive or dysplastic margins were found in the initial specimen in 20.0%of the cases.On multivariate analysis,there was no significant difference between RFS(hazard ratio(HR)1.379,p = 0.361)or DFS(HR 1.452,p = 0,183)of those with mild dysplasia and those with negative margins.However,patients with mild dysplasia who did not undergo re-excision demonstrated significantly worse RFS(HR 2.286,p = 0.010)and DFS(HR 2.070,p = 0.014)than those with negative margins.[Conclusions]There appears to be a correlation between initial mild dysplastic margins that are not subjected to re-excision and inferior RFS and DFS.Additional attention should be drawn to mild dysplasia at the initial margin in OSCC,and extended excision is suggested.TwoThe influence of pattern of invasion on the prognosis of oral squamous cell carcinoma[Objective]To evaluate the prognostic value of worst pattern of tumour invasion(WPOI)and biopsy pattern of invasion(BPOI)of patients with early-stage oral squamous cell carcinoma(OSCC),To assess the ability of CD1a+DC and BPOI to predict WPOI preoperationly.To evaluate OSCC patients with only micrometastasis and inferior worst pattern of invasion(WPOI),as well as those with ?/?levels of micrometastasis,need postoperative radiotherapy is unclear.[Methods]1.This retrospective study included 444 patients with pTl-2N0M0 OSCC who had undergone surgical treatment only in the Department of Oral and Maxillofacial Surgery,Nanjing Stomatology Hospital from 2005 to 2014.The value of WPOI and BPOI in predicting prognosis and the ability of CD1a+DC in predicting the WPOI preoperationly were assessed.Statistical analyses were performed using the ?2 test,Kaplan-Meier method,and Cox regression model.Predictors of outcome were identified using multivariate analysis.2.OSCC patients(n=311)with negative(n=247),only micrometastasis(n=44)and macrometastasis(n=20)were detected and selected by HE staining.Micrometastasis was re-assessed using immunohistochemical staining of cytokeratin(CK)in HE negative patients to find out the false negative cases.The incidence and clinical features of the primary tumour in relation to the only micrometastatic status,and the value of the postoperative radiotherapy on the only micrometastasis patients were evaluated.[Results]1.The outcome(including overall survival,recurrence-free survival and disease-free survival 1)of patients with WPOI 4-5 is worse than WPOI 1-3(p=0.002,p=0.022,p=0.002,respectively).However,BPOI was not significantly associated with OS,RFS,or DFS(p=0.979,p=0.422,p=0.440,respectively).But among the 80 BPOI type 4 cases,their matching WPOI were all type 4-5.And WPOI 4-5 was significantly associated with a deeper invasion which was also significantly associated with OS,RFS,or DFS.The areas under ROC were 0.769 for CD1a+DC/HP,the sensitivity and specificity were 73.0%and 74.4%,the optimal threshold values was 21.833.When combined the two methods the sensitivity and specificity were 88.2%and 84.6%.2.Among the negative lymph node cases(n=247),the positive rate of CK was 4.94%(n=12).Patients with only micrometastasis had higher T stage and inferior WPOI than patients without micrometastasis,but they had longer overall survival(OS),metastasis-free survival(MFS)and Disease-free survival(DFS)than macrometastasis patients.Micrometastasis is an independent diagnostic indicator for MFS.However,the survival time of only micrometastasis patients with or without postoperative radiotherapy was comparable,even in patients with inferior WPOI.Radiotherapy,however,may only benefit patients with IV/V levels of micrometastasis.[Conclusions]The WPOI was significant indicators of prognosis.BPOI type 4 indicates WPOI type 4-5 invasion and is predictive of a poor outcome,thus more extensive resection should be considered for such patients.The number of CD1a +DC cells infiltrating the front of tumor biopsy tissue as an observational index which can improve the accuracy of assessing WPOI when combined with BPOI.There appears to be a correlation between WPOI and micrometasitasis.Postoperative radiotherapy is dispensable for micrometastasis.ThreeEffects of Different WPOI on Recruitment of Tumor-Associated Neutrophils[Objectives]To study infiltrating tumor-associated neutrophils(TANs)in OSCC microenvironment and their prognostic value;the distribution of TANs in different WPOI;effects of different types of WPOI on recruitment of TANs.[Methods]OSCC patients with complete follow-up data who received primary surgical treatment in Affiliated Stomatological Hospital of Nanjing University Medical School from 2005 to 2010 were chosen for the study.Difference of numbers of neutrophils in OSCC tissue between WPOI 1-3 and WPOI 4-5 were determined with immunohistochemistry and flow cytometry.The ability of IL-7 and CXCL5 to recruit neutrophils in different types of WPOI was accessed with immunohistochemistry.[Results]Univariate analysis showed that neutrophil density has significant influence in OS(p=0.0003),RFS(p=0.017)and DFS(p=0.003);CD15+ neutrophils at the invasion front were significantly different between WPOI 1-3 and WPOI 4-5(p= 0.0132,p=0.0027).Flow cytometry showed the same result.This difference may be related to high expression of IL-17 and CXCL5 at WPOI 4-5 invasion front.[Conclusions]Though the density of CD15+ neutrophils was not an independent prognostic factor of OSCC,it influenced the survival rate of OSCC patients.In WPOI 4-5,TANs increased and mainly distributed in the invasion front,especially in worse pattern of invasion.CXCL5 and IL-17 participated in recruitment of TANs.