| Background&Objectives:Cancer has become a major public health problem that seriously threatens the health of Chinese residents.The Health China Action-Cancer Prevention and Control Implementation Plan(2019-2022)has proposed the core actions of controlling risk factors,application of cancer informational big data,improving cancer prevention and treatment capacity,promoting early diagnosis and treatment of cancer and other core action contents.China is a high incidence area for digestive system tumors,among which gastric cancer is the main point of cancer prevention and control.However,when gastric cancer is diagnosed,most of them have local infiltration and distant metastasis,and the mortality rate is high.Therefore,how to prevent and control gastric cancer is of great significance.Similar to many tumors,gastric mucosal lesions are usually preceded by a chronic inflammatory process,i.e.,the scientific problem of inflammation-to-cancer.The Correa’s Cascade hypothesis suggests that the development of intestinal gastric cancer(Lauren’s type)generally follows the following evolutionary pattern:normal gastric mucosa→ chronic non-atrophic gastritis(CNAG)→chronic atrophic gastritis(CAG)→intestinal IM→atypical hyperplasia of gastric mucosa(DYS)→carcinoma of gastric mucosa(ML),indicating that the development of gastric cancer is a multi-stage chronic evolutionary process,which provides a basis for the prevention and control of gastric cancer.Since inflammation is controllable and cancer is uncontrollable,the early management of gastric mucosal lesions related to the Correa’s Cascade hypothesis has become an important part of gastric cancer prevention and control.Early management of gastric mucosal lesions has become the focus of gastric cancer prevention and control.In China,not only the incidence of gastric cancer is high,but also the prevalence of H.pylori infection,which is a major cause of many upper gastrointestinal diseases,including gastric cancer,and is considered to be the Correa’s Cascade hypothesis.H.pylori infection is a major cause of many upper gastrointestinal diseases,including gastric cancer,and is considered to be the initiator of the Correa’s Cascade hypothesis of gastric mucosal lesions.Therefore,the eradication of H.pylori is an important starting point for the management of gastric mucosal lesions related to the Correa’s Cascade hypothesis,a concrete manifestation of controllable inflammation,and a theoretical basis for improving the ability to prevent and treat gastric cancer.It is also a theoretical basis for improving the prevention and treatment of gastric cancer.It is worth pointing out that the Correa’s Cascade hypothesis describes the step-by-step cross-linking and chronological evolution of the relevant gastric mucosal lesion stages.The progression pattern of Correa’s Cascade is a scientific hypothesis on the pattern of gastric carcinogenesis by observing the pathological changes of gastric mucosal lesions over time in a high-risk cohort of gastric cancer,which is a qualitative study.Therefore,the scientific validity of the Correa’s Cascade hypothesis needs to be supported by more real-world data.In addition,the magnitude of the risk of gastric cancer for each gastric mucosal lesion in the Correa’s Cascade hypothesis is not clear,and the risk of H.pylori triggering the Correa’s Cascade hypothesis is not clear.The targets and molecular events of H.pylori triggering the development and progression of gastric mucosal lesions related to the Correa’s Cascade hypothesis are still poorly understood.The ERM family is an important linking protein between the cytoplasmic membrane and the cytoskeleton and a major component of the cellular microvilli,including the following four members:ezrin,radixin,moesin,and merlin-like proteins.They are involved in the regulation of cell membrane structure and cell morphology by affecting the cytoskeleton,have important roles in maintaining cellular morphology and cell motility,and act as cellular cortical signaling integrators,participating in cell polarization,invasion,migration and adhesion processes by communicating membrane protein-mediated signaling.In recent years,it has been found that some ERM family(ezrin/radixin/moesin,ERM family)protein members are not only closely related to the pathogenesis of various pathogenic microbial infections,but also play an important role in the progression of digestive system tumorigenesis.Therefore,it is necessary to investigate the role of ERM family proteins in the inflammatory-cancerous transformation of gastric mucosal lesions associated with the Correa’s Cascade hypothesis caused by H.pylori Therefore,it is necessary to investigate the role of ERM family proteins in the process of inflammation-cancer transformation of gastric mucosal lesions related to the Correa’s Cascade hypothesis of H.pylori,in order to provide a reference for the prevention and control of gastric cancer and the molecular mechanism of gastric cancer progressionMethods:1.relationship between H.pylori and Correa’s Cascade(1)A cross-sectional survey study was designed to extract information on personal characteristics,clinical information,gastroscopy records and pathological descriptions of all those who underwent gastroscopy for various reasons at the Gastroenterology Endoscopy Center of the First Affiliated Hospital of Nanchang University during a consecutive 5-year survey interval from January 1,2015 to December 31,2019.(2)The data were cleaned and organized according to the screening criteria,and the total detection rate of gastric mucosal lesions related to Correa’s Cascade hypothesis was calculated based on the information of gastroscopic biopsy pathology,and the detection rate of gastric mucosal lesions related to Correa’s Cascade hypothesis was calculated in different age groups.(3)The co-occurrence(co-occurrence of two to two)distribution table of Correa’s Cascade hypothesis-associated gastric mucosal lesions detected in any one subject over 5 years was made,and the co-occurrence rate of gastric mucosal lesions was calculated to analyze the stepwise cross-linking and time-series evolution of Correa’s Cascade hypothesis-associated gastric mucosal lesions.(3)To analyze the stepwise cross-linking and temporal evolution of gastric mucosal lesions related to Correa’s Cascade hypothesis(4)To calculate the H.pylori positive rate based on the information of gastroscopic biopsy pathology detection,to analyze the difference of H.pylori positive rate in Correa’s Cascade associated gastric mucosal lesions,and to explore the association between the degree of H.pylori infection and the degree of Correa’s Cascade associated gastric mucosal lesions.(5)To analyze the evolving trend of H.pylori positivity rate and detection rate of gastric mucosal lesions associated with Correa’s Cascade hypothesis based on 5 consecutive years of gastroscopic data.2.Preliminary investigation of the mechanism of ERM family in Correa’s Cascade(1)Gene expression profile information data(GSE106656 and GSE11631)of Correa’s Cascade related gastric mucosal lesions were obtained from the GEO database,and the differences in ERM family expression in different gastric mucosal lesions were analyzed using the official website GEO2R(2)Obtaining transcriptome data and clinical information of gastric cancer from TCGA,and analyzing the expression differences of ERM family in gastric cancer and paraneoplasia using and related program package(edgR and DESeq2).(3)To analyze the differences of ERM family in different gastric cancer tumor staging(Lauren staging:3 types of intestinal gastric cancer,diffuse gastric cancer and mixed gastric cancer),different genders(male and female)and different tumor stage(Stage Ⅰ-Ⅳ)in the Oncomine database.(4)GEPIA was used to analyze the differences in the expression of ERM protein family genes in gastric cancer tissues,paraneoplastic tissues and normal gastric tissues,and to analyze the relationship between the expression levels of ERM protein family genes and the prognosis of gastric cancer.(5)The relationship between ERM protein family gene expression levels and gastric cancer prognosis was supplemented and validated by using Kaplan-Meier plotter.(6)The results of differential expression analysis of ERM protein family genes in gastric mucosal lesions based on Correa’s Cascade hypothesis were further analyzed by using R language to achieve gene function enrichment analysis(GSEA).(7)Based on the scientific problem of inflammation-cancer transformation,the immune infiltration analysis was further performed by TIMER database,combining the molecular mechanism of H.pylori pathogenesis,the molecular biological characteristics of ERM protein family and the gene function enrichment results.3.Expression and significance of moesin of ERM family in gastric mucosal lesions(1)26695,ATCC43504(i.e.NCTC11637),7.13,PMSS1,G27 and CCS9803 were selected as standard or model strains of H.pylori(cagAPMSSl>cagA7.13),and another immortalized human embryonic gastric mucosal epithelial cells(GES-1),and human gastric gland HGC-27 and BGC-823,low-differentiated human gastric gland cancer cells AGS and SUN-1,and moderately differentiated human gastric gland cancer cells SGC-7901 were selected as experimental cells.(2)Correa’s Cascade hypothesis-related gastric mucosal lesions were selected from tissue wax blocks preserved in the pathology department,and the lesions were CNAG(containing mild and severe,i.e.,MCNAG and SCNAG),CAG,IM,DYS,and ML,all containing information on H.pylori infection.(3)Twelve cases of human fresh gastric cancer tissues and corresponding paracancerous tissues were selected as validation supplements.(4)qRT-PCR assay to detect moesin mRNA expression in each experimental cell(5)Western blot assay to detect the changes of moesin expression levels in each cell line after incubation with H.pylori.(6)Western blot and immunohistochemical staining to detect the expression of moesin protein in gastric cancer and paraneoplastic tissues.(7)Immunohistochemical staining was performed to detect the expression of moesin protein in gastric mucosal lesions associated with Correa’s Cascade hypothesis of combined H.pylori infection.Results:1.Relationship between H.pylori and Correa’s Cascade(1)During the 5-year survey interval from 2015 to 2019,a total of 399,059 cases were initially screened for gastroscopy at the Gastroenterology Endoscopy Center of the First Affiliated Hospital of Nanchang University for various reasons.In combination with the established screening criteria,87,029 cases were further excluded from the data of 399,059 cases by R-language data cleaning.Finally,a total of 312030 cases of gastroscopy subjects meeting the screening criteria were obtained in this survey study.By year,the number of gastroscopy cases from 2015 to 2019 were 58172,61292,65661,63027,and 63878 cases,respectively(2)The results of data screening revealed that a total of 171,974 cases of gastroscopic mucosal biopsy pathology were performed in the data of 312,030 subjects,and the total biopsy rate of subjects during gastroscopy was about 55.1%(171,974/312030).If divided by year,the number of gastroscopy biopsy cases from 2015 to 2019 were 20387,29136,38556,41406,and 42489,corresponding to biopsy rates of 35%,48%,59%,66%,and 67%,respectively.(3)Among the 171974 patients who underwent gastric mucosal biopsy pathology,the youngest was less than 10 years old and the oldest was 96 years old,with an overall mean age of(50.9±12.9)years.The results of gastric mucosal biopsy pathology showed that two or more Correa’s Cascade hypothesis-related gastric mucosal lesions could exist simultaneously,and each related gastric mucosal lesion was not independent of the other,and some of them could be crossover or overlap The related gastric mucosal lesions are not independent,and some of them can be crossover,overlap or co-occurrence.(4)There was a trend in the detection rate of gastric mucosal lesions associated with Correa’s Cascade hypothesis in all age groups.Specifically,CNAG was commonly detected in all age groups,mainly in adolescents,and its detection rate tended to decrease with increasing age,while CAG+,DYS,and ML showed an increasing trend with increasing age.The trend of these gastric mucosal lesions with age is highly consistent with the sequence of associated gastric mucosal lesions described by Correa’s Cascade hypothesis.(5)In different gastric mucosal lesions,the degree of closeness of association among the gastric mucosal lesions is not completely uniform,and the lesions are not juxtaposed but may be sequential and have a preferential direction of development,and the development trend of CNAG,CAG+(ie,CAG+IM),DYS,and ML is progressive in a stepwise correlation,i.e.,the lesions are sequential(stepwise),and the existing lesions develop preferentially toward a particular gastric mucosal lesion(cross-linking),and in pathophysiological(without external intervention),this direction of progression is irreversible.(6)The probability of CNAG combined ML was 0.8%,CAG+combined ML was 1.1%,and DYS combined ML was 11.0%.(7)Among the 171,977 gastroscopic mucosal biopsies and pathological examinations,a total of 122,735 cases of H.pylori infection status were examined by special staining,and the detection rate was as high as 71.4%(122,735/17,977),which has raised the clinical awareness and attention to H.pylori infection.The overall positive rate of H.pylori in the last 5 years was 31.0%.(8)The positive rate of H.pylori infection in the Correa’s Cascade-associated gastric mucosal lesion group was higher than that in the group without Correa’s Cascade-associated gastric mucosal lesion(P<0.001),and the percentage of each Correa’s Cascade-associated gastric mucosal lesion increased with the increase of the grade or degree of H.pylori infection.The positive rate of H.pylori infection in the Correa’s Cascade-associated gastric mucosal lesion group was higher than that in the group without Correa’s Cascade-associated gastric mucosal lesion(all P<0.001),suggesting that the degree of gastric mucosal lesion was related to the degree of H.pylori infection.Further analysis showed that the differences in the degree of Correa’s Cascade associated gastric mucosal lesions were all statistically significant in relation to the degree of H.pylori infection status(grading and classification),except for the DYS lesions(all P<0.001).(9)There was an association between H.pylori infection and each Correa’s Cascade associated gastric mucosal lesion and degree of lesion,and the percentage of each Correa’s Cascade associated gastric mucosal lesion degree increased with the increase of the graded or graded degree of H.pylori infection,and the trend of change was basically the same in the last 5 years.Overall,the evolution of H.pylori positivity,DYS and ML showed a consistent trend of decreasing year by year.The trend changes in the detection rate of gastric mucosal lesions associated with Correa’s Cascade hypothesis in the last 3 years(2017-2019)all showed a slowing trend.2.Preliminary investigation of the mechanism of ERM family in Correa’s Cascade(1)In the GEO data analysis,it was found that the four members of ERM protein family genes were not different in CNAG vs CAG and CNAG vs IM,while only moesin gene expression was found to be upregulated in both CNAG vs ML and CAG vs ML,and the differences were statistically significant(all P<0.05).(2)In TCGA-STAD data analysis,only moesin gene was found to be differentially expressed among ERM protein family genes,and its expression level in cancer tissues was higher than that in gastric cancer parietal tissues,and the difference was statistically significant(P<0.05).(3)The combined analysis of TCGA-STAD and GTEx data in GEPIA revealed that only moesin and merlin genes were differentially expressed in the ERM protein family,and their expression levels were higher in cancer tissues than in gastric paraneoplastic tissues and normal gastric tissues,with statistically significant differences(P<0.05 for both).(4)Among the different clinical stages,only moesin gene was found to be statistically significant(P<0.05)among the ERM protein family genes,and the expression levels in stages II-III were significantly higher than those in stage I.(5)Among the different gastric cancer types(Lauren typing),the expression of moesin and merlin genes among ERM protein family genes was found to be different and stable,and the expression levels were higher than those of paraneoplastic tissues(both P<0.05).(6)High expression of moesin gene predicted poor overall prognosis(OS and DFS)of gastric cancer,while no correlation was found between the expression level of merlin gene and prognosis of gastric cancer.(7)The expression level of moesin gene was correlated with tumor grade and T stage of gastric cancer patients,but not with gender,age and N/M stage.(8)The expression level of Moesin gene in gastric cancer tissues was not only higher than that of paraneoplastic group,but also higher than that of normal gastric tissues.In addition,its expression level in gastric cancer tissues with combined H.pylori infection was also higher than that in gastric cancer tissues without H.pylori infection.(9)Gene enrichment analysis showed that moesin genes in gastric cancer were mainly associated with cytokine-cytokine receptor interaction pathway,Janus kinase-signal transducer and activator of transcription(JAK-STAT)pathway,cell adhesion molecule pathway,calcium ion signaling pathway,Toll-like receptor signaling pathway and extracellular matrix receptor(10)The moes in gastric cancer is related to the JAK-STAT pathway,cell adhesion molecule pathway,calcium signaling pathway,Toll-like receptor signaling pathway and extracellular matrix receptor pathway.(10)The moesin genes in gastric cancer were closely correlated with JAK1,STAT5,ZEB1 and ZEB2,and positively correlated with NF-κB and IL-6(11)In TIMER analysis,moesin genes were found to have positive correlation with most immune cells in gastric cancer tissues except B cells,and the correlation coefficients were in descending order of dendritic cells,macrophages,neutrophils,and T cells.3.Expression and significance of ERM family protein moesin in gastric mucosal lesions(1)The expression level of moesin protein was higher in both GES-1/AGS cells after strain addition compared to the no-strain group(P<0.01)(2)Compared with the GES-1/AGS group without strain addition,the moesin protein expression was higher in both the GES-1/AGS+PMSS1 and GES-1/AGS+7.13 groups,and the difference between the two was statistically significant(P<0.01).Moreover,the expression level of moesin protein was higher in the GES-1/AGS+PMSS1 group than in the GES-1/AGS+7.13 group,and the difference between the two was statistically significant(P<0.05),indicating that the expression of moesin protein may be partly through the cagA of the strain(cagAPMSS1>cagA7.13)(3)Compared with the GES-1/AGS group without 7.13,the moesin protein expression was higher in both the GES-1/AGS+7.13 group and the GES-1/AGS+cagA knockdown 7.13 group,and the difference between the two was statistically significant(P<0.01).Moreover,the expression level of moesin protein in the GES-1/AGS+cagA knockdown 7.13 group was lower than that in the GES-1/AGS+7.13 group but higher than that in the GES-1 group without 7.13,and the difference between the two groups was statistically significant(P<0.05),further confirming that cagA of the strain could stimulate GES-1/AGS cells to moesin protein expression level was increased.(4)The expression of moesin protein in 12 pairs of clinical gastric cancer and paired tissue samples was detected by Western blot method,and the results showed that 83.3%(10/12)of the moesin protein was expressed at a higher level in gastric cancer tissues than in paracancerous tissues,and the difference was statistically significant(P<0.01),and the detection using immunohistochemical staining also showed that The expression level of moesin protein in gastric cancer tissues was higher than that in paraneoplastic tissues,and the difference was statistically significant(P<0.01)(5)The expression of moesin protein in gastric mucosal lesions associated with Correa’s Cascade hypothesis was detected by immunohistochemical staining,and the results showed that moesin protein expression in gastric mucosal lesions associated with Correa’s Cascade hypothesis was statistically significant in both inflammatory cells and glandular cells.The difference in the expression of moesin protein in Correa’s Cascade hypothesis-related gastric mucosal lesions was statistically significant(P<0.01),and the expression in ML was much higher than that in other gastric mucosal lesion tissues,which was significant in inflammatory cells(6)Immunohistochemical staining was used to detect the expression of moesin protein in gastric mucosal lesions associated with Correa’s Cascade hypothesis of combined H.pylori infection,and the results showed that moesin protein was expressed in both inflammatory cells and glandular cells in Correa’s The difference in the expression of moesin in Correa’s Cascade hypothesis-associated gastric mucosal lesions was statistically significant,and the expression in ML tissues was much higher than that in other gastric mucosal lesions.Moreover,the expression of moesin protein in both inflammatory and glandular cells was higher in gastric mucosal lesions associated with combined H.pylori infection Correa’s Cascade hypothesis than in gastric mucosal lesions associated with Correa’s Cascade hypothesis without H.pylori infection lesions,as evidenced by inflammatory cellsConclusions:1.The progression of gastric mucosal lesions in the Correa’s Cascade hypothesis is consistent with real-world evidence,and this hypothesis of gastric carcinogenesis pattern is scientifically valid.The Correa’s Cascade hypothesis,in which the risk of combining different stages of gastric mucosal lesions with cancer varies,is a definite guide for gastric cancer prevention and control.2.H.pylori infection is related to the gastric mucosal lesions associated with the Correa’s Cascade hypothesis,and the trends of the two are basically the same.Therefore,the control of H.pylori infection under the guidance of the Correa’s Cascade hypothesis has positive significance for the prevention of gastric cancer.3.The ERM family protein moesin was highly expressed in gastric mucosal lesions related to the Correa’s Cascade hypothesis,especially at the stage of gastric mucosal carcinoma,which was associated with H.pylori and its cagA gene4.High expression of moesin is associated with gastric cancer progression and predicts poor prognosis of gastric cancer,so abnormally high expression of moesin may play a pro-cancer role and is expected to be a diagnostic biomarker or therapeutic intervention target for gastric cancer. |
PDF Full Text Request