Comparison Of PLCE1 Protein Expression And Its Effect On Survival In High And Low Incidence Esophageal Squamous Cell Carcinoma

1 Research background and purposeEsophageal cancer(EC)is one of the most common malignant tumors in the world.It is located in the fourth place of cancer death in China,ESCC(esophageal squamous cell carcinoma)and esophageal adenocarcinoma(EAC).In China,97%of esophageal cancers are esophageal squamous cell carcinoma.The two epidemiological characteristics of esophageal cancer are the high and low incidence areas and the family aggregation phenomenon with obvious regional distribution difference.The pathogenesis of esophageal cancer,especially the molecular mechanism of esophageal cancer,has attracted great attention in recent years.It has been proved that esophageal cancer is the result of the interaction of environmental factors and genetic factors.However,some studies show that in the high incidence area,the exposure time of esophageal cancer patients to carcinogenic environment is much longer than that in the low incidence area,and the positive rate of family history is also higher than that in the low incidence area.The patients in high incidence areas are greatly affected by environmental carcinogenic factors,while the patients in low incidence areas are relatively less affected by environmental carcinogenic factors,so why do people in low incidence areas also have esophageal cancer?Is it because the proportion of genetic factors in patients with esophageal cancer in high and low incidence areas is different?Or because the pathogenesis of esophageal cancer patients in high and low incidence areas is different?PLCE1 is the first reported susceptibility gene.In this study,we compared the expression of PLCE1 protein in patients with esophageal cancer in high and low risk areas to further determine whether the genetic factors of high and low risk esophageal cancer patients are consistent.PLCE1 gene is located on chromosome 10q23,with a length of 334.3kb and a protein molecular weight of 258000,including 2302 amino acids.PLCE1 encodes phospholipase C epsilon-1(PLCE1),which hydrolyzes phosphatidylinositol 4,5-diphosphate to 1,2-diacylglycerol and inositol 1,4,5-triphosphate,thus promoting intracellular signal transduction.The PLCE1 gene has been shown to be associated with susceptibility to esophageal and other cancers.Our team applied genome wide association study(GWAS)technology to report that PLCE1 gene is the susceptible gene of esophageal squamous cell carcinoma for the first time in the world.In this study,we compared the expression of PLCE1 protein in patients with esophageal cancer in high and low risk areas,and explored the molecular mechanism of environment genetic interaction and the influence of prognosis.2 Materials and methods2.1 Research objectThe data of this study were selected from the 500000 cases of esophageal and cardiac cancer clinical information database of the State Key Laboratory for prevention and treatment of esophageal cancer jointly established by the provincial and Ministry of the First Affiliated Hospital of Zhengzhou University.According to the inclusion criteria,the final subjects included were 117186 cases of esophageal squamous cancer patients,with the diagnosis time between March 1970 and November 2016,including 73646 cases in high incidence area,43540 cases in low incidence area,and 73 men,916 cases were 22-90(60.2±9.2)years old;43270 women were 25-90(61.1 ± 9.2)years old;the ratio of male to female was 1.7:1.2.2 Clinical diagnosis and pathological information collectionThe clinical diagnosis,treatment and pathological information of 117186 patients with esophageal squamous cell carcinoma in this study are all from the inpatient medical records of 500000 cases of esophageal and cardiac cancer in the State Key Laboratory of esophageal cancer prevention and control jointly established by the province and Ministry of the First Affiliated Hospital of Zhengzhou University,including the basic information of patients,such as name,age,gender,detailed family address,contact person,contact information,pathology and follow-up Information pathology is also clear,and check and supplement the patient's pathology and clinical diagnosis and treatment information in each relevant hospital to ensure the accuracy of the patient's information.2.3 issue sample collection,treatment and PLCE1 immunohistochemical stainingMost of the tissue samples collected by the research team come from:the remaining samples after routine collection and diagnosis of the resected samples from various hospitals.Firstly,the case of esophageal squamous cell carcinoma was confirmed.Then,the patients and their families were communicated before the operation,and the informed consent was signed for specimen collection.Then,the remaining samples were transferred to the laboratory for storage in the liquid nitrogen tank after the diagnosis of the routine samples collected in the hospital.Finally,the specimens were fixed,taken,embedded in paraffin and stained with immunohi stochemistry.2.4 follow-upIn this study,all ESCC patients were followed up by telephone inquiry,household survey and village medical consultation,and their living or death and other detailed living conditions,time of death,last follow-up time,cause of death,follow-up person,follow-up time were recorded.The patients were followed up every 3 months in the first year after discharge and every year thereafter.The end point of follow-up was the time of death or until November 2019.Survival time is the date of death or last follow-up minus the date of diagnosis.2.5 Method?The clinical and pathological information of 73646 and 43540 patients with esophageal cancer from high and low incidence areas was analyzed,including basic information such as sex,age,family history,high and low hair area,etc.,and pathological information,such as tumor site,tumor length,degree of separation,Cut-edge net,lymph node transfer negative positive,TNM staging,etc.?Immunized tissue and cancer tissue samples of 375 patients with esophageal cancer were immuno-grouped,and by detecting positive expression of PLCE1,the relationship between PLCE1 expression and clinical pathological characteristics of esophageal cancer patients was analyzed,and the final analysis was high,The difference of PLCE1 expression in cancerous tissue in patients with esophageal cancer in low-hair region is of clinical significance to patients.?This study data is analyzed using SPSS 21.0.Sex,age,high and low hair region,smoking history,history of alcohol consumption,family history of tumors,tumor site,tumor length,cut-off net,lymph node metastasis negative positive distribution using the test,degree of separation,TNM phased and treatment application of rank and test;The effect of kaplan-Meier curve and Log-rank test is compared on survival,and the main factors of survival are analyzed by using the multi-factor Cox risk regression regression model method,and the test level was ?=0.05.3 Result3.1 Clinicopathological features of esophageal squamous cell carcinoma in high and low incidence areasAmong the 117186 ESCC patients,736 were in high incidence area,46(75.0%),435 in low incidence area,40(25.0%),and the ratio of high and low incidence area was 1.7:1.There were significant differences in the distribution of sex,age,drinking history,smoking history,cancer family history,tumor location,tumor length,differentiation degree,cutting edge,lymph node metastasis,TNM stage and treatment mode between the high and low incidence areas(P<0.05).3.2 Single factor analysis of the influence of high and low incidence areas on the survival of patients with esophageal squamous cell carcinomaIn the high incidence area,the survival curves of sex,age,smoking,drinking,family history,tumor location,tumor length,clean margin,differentiation degree,lymph node metastasis,TNM stage and treatment were statistically significant(P<0.05);In the low incidence area,sex,age,smoking,drinking,tumor family history,tumor location,tumor length,differentiation degree,lymph node metastasis,TNM stage and treatment mode of survival curve were statistically significant(P<0.05).3.3 Analysis of Cox risk ratio regression model for esophageal cancer patients in high and low risk areasCox risk proportional regression model was used to analyze the significant variables of single factor analysis results in high and low incidence areas.In high incidence areas,age,gender,smoking history,tumor family history,tumor location,tumor length,differentiation degree,lymph node metastasis,yin and Yang,TNM stage were independent factors affecting survival of esophageal squamous cell carcinoma patients;in low incidence areas,age,gender,tumor family history Tumor location,tumor length and diameter,differentiation degree,lymph node metastasis and TNM stage are independent influencing factors for survival of ESCC patients.3.4 Comparison of PLCE1 protein expression and its effect on survival in high and low incidence esophageal squamous cell carcinomaThe positive expression rate of PLCE1 in the adjacent and cancer tissues of esophageal cancer patients in the high and low incidence areas showed an upward trend.In high and low incidence areas,PLCE1 positive and negative esophageal cancer patients had similar survival,and weak and strong positive esophageal cancer patients had similar survival.4 Conclusion4.1 The overall survival of esophageal squamous cell carcinoma patients in high incidence area is better than that in low incidence area.4.2 Low incidence area,male,?60,smoking,negative family history,neck+upper thoracic segment,tumor length>4.0,positive lymph node metastasis,low differentiation,TNM stage are independent risk factors for poor prognosis of esophageal cancer patients.4.3 The positive expression rate of PLCE1 in the high and low incidence areas,adjacent to esophageal cancer and cancer tissues all showed an upward trend,suggesting that PLCE1 may play an important role in the carcinogenesis of esophageal cancer.4.4 In the high and low incidence areas,the expression of PLCE1 was not related to the survival of ESCC patients.