The Value Of Carcinoma-associated Fibroblast Factors Connexin 43 And ?-SMA In Pathological Diagnosis Of Early Invasive Carcinoma Of Oral Squamous Cells

BackgroundOral squamous cell carcinoma(OSCC)is a common malignant tumor in oral and maxillofacial region,which seriously affects human health and quality of life.The incidence of oral squamous cell carcinoma has increased by 50%in the past ten years.Mostly,it occurs in males over the age of 40.Most of the OSCC diagnosis progresses to?~?stage,5-year survival rate was only 53%,in order to improve the quality of life of patients,the early diagnosis of OSCC is particularly important.The higher the degree of dysplasia of oral mucosal squamous epithelium,the higher the potential malignancy.Therefore,a clear and early diagnosis of oral squamous epithelial severe dysplasia(OSESD),oral squamous cell carcinoma in situ(OSCCIS),and oral squamous cell carcinoma in situ with micro-infiltration(OSCCISM)are closely related to the treatment and prognosis of patients.In recent years,more and more researches have made on tumor microenvi-ronment,and the study of tumor-associated fibroblasts,the main component of tumor microenvironment,have also made a great progress.It can activate transforming growth factor-?to regulate keratinocyte adhesion and secrete a wide range of cytokines,such as growth factors,chemokines,hormones,and play an important role in tumor growth,metastasis,invasion and tumor-induced inflammation and immune evasion.At the same time,the increase of CAFs indicates poor prognosis.Many studies suggest that CAFs are transformed by the normal fibroblasts in the tumor stroma stimulated by the relevant factors secreted by tumor cells.some studies have shown that CAFs express?-SMA,while normal oral fibroblasts do not express?-SMA,and CAFs also express Connexin 43.In theory,normal fibrous tissue is converted to CAFs only when cancerous cells infiltrate subepithelial fibrous tissue from the epithelium in situ.Based on this,we speculate that the detection of relevant indicators of CAFs can help determine the early infiltration of cancer.This subject has collected different types of oral mucosal squamous intraepithelial lesions,early invasive carcinoma and invasive carcinoma.Immunohistochemistry was used to analyze the single staining and double staining of CAFs factors Connexin 43 and?-SMA to judge whether these indexes have the value of assisting in judging the early invasion of squamous cell carcinoma.ObjectiveThe use of immunohistochemical staining and double staining methods to detect the expression of Connexin 43 and?-SMA in different types of squamous epithelial lesions of the oral mucosa,and to explore whether they have the value of assisting in judging the early invasion of squamous cell carcinoma.Methods1.Collected 197 cases of oral mucosal lesion resection in patients from 2011 to2014 in the First Affiliated Hospital of Zhengzhou University.We selected 54 OSESD,58 OSCCIS,and 75 OSCCISM/OSCC.In addition,10 chronic inflammation of oral mucosa(CIOM)as the control group;2.The immunohistochemical single staining(Connexin 43),double-staining(?-SMA and ERG,which can remove vascular smooth muscle)and double-staining(Connexin 43 and?-SMA,which can improve the detection rate of CAFs)were used in 10 CIOM,54 OSESD,58 OSCCIS,and 75 OSCCISM/OSCC to detect the expression of Connexin 43 protein and?-SMA protein,as well as the co-expression of Connexin 43 and?-SMA protein,to analyze the relationship among Connexin 43,?-SMA,and the CAFs they represent with the clinicopathological diagnosis of oral squamous epitheliopathy.3.Follow-up:Follow-up by phone to patients with OSESD and OSCCIS and observe the progress of the disease.4.Statistical analysis:SPSS21.0 statistical software was used for data analysis and?~2 test,P<0.05 was considered statistically significant differences.Results1.Connexin 43 single stain:The positive rates of Connexin 43 in the stroma of CIOM,OSESD,OSCCIS and OSCCISM/OSCC were 10.00%,11.11%,43.10%and61.33%respectively.Except for the difference between CIOM and OSESD was not statistically significant,the differences between the other groups were statistically significant(P<0.05).2.?-SMA and ERG double staining:Except for the vascular area,the positive rates of?-SMA were 0.00%,9.30%,34.48%,68.00%in the stroma of CIOM,OSESD,OSCCIS and OSCCISM/OSCC.Except for the difference between CIOM and OSESD was not statistically significant,the differences between the other groups were statistically significant(P<0.05).3.Connexin 43 and?-SMA double staining:The positive rates of CAFs in the stroma of CIOM,OSESD,OSCCIS and OSCCISM/OSCC were 10.00%,16.67%,55.17%and 85.33%.Except for the difference between CIOM and OSESD was not statistically significant,the differences between the other groups were statistically significant(P<0.05).4.In OSCCISM/OSCC,stroma cells surrounding infiltrating carcinoma were clearly positive for CAFs factors.Among OSESD and OSCCIS follow-up results,Patients with positive CAFs in stroma are more likely to progress to OSCC than patients with negative CAFs.Conclusions1.This study demonstrated the significant differences in the expression of CAFs,which expressed in Connexin 43 and?-SMA,in different degrees of oral mucosal squamous epithelial lesions.2.According to the follow-up results of OSESD and OSCCIS patients,the increase of CAFs is a poor prognostic indicator,suggesting an increased probability of tumor progression.3.In determining whether there is partial infiltration of OSCCIS,if the expression of CAFs were positive in the surrounding stroma of the suspicious cancer,the diagnosis is more supportive of invasive cancer.Therefore,Connexin 43 and?-SMA have some value in the diagnosis of oral squamous cell carcinoma and early invasion of squamous cell carcinoma.