| Purpose:In this study,we used 16 S rRNA full-length sequencing technology to analyze the differences in the diversity and composition of intra-tumor flora in cervical,endometrial and ovarian cancers,to initially explore the effect of age on the distribution of intra-tumor flora,and to analyze the correlation between the diversity of intra-tumor flora and clinical biochemical indices and tumor markers,to provide a new protocol for the diagnosis and treatment of gynecological malignancies.Methods:1.Inclusion of study subjects and collection of tumor tissues: with the informed consent of patients,tumor samples were collected from patients who underwent gynecological malignant tumor resection at the Second Affiliated Hospital of Nanchang University hospital from September 2020 to November 2022,and were divided into three groups according to the type of pathology.cervical cancer group(Group C),endometrial cancer group(Group E),and ovarian cancer group(Group O),and the collected tumor tissues were stored in-80℃ environment for backup.2.Collection of data from enrolled patients: values of hemoglobin,leukocytes,platelets,Neutrophil ratio,Lymphocyte ratio,albumin and glycoantigen-125 were collected from the three study groups.3.16 S r RNA full-length sequencing of bacterial DNA within tumors: A total of90 patients undergoing gynecologic malignancy resection were included in this study,divided into three groups: 30 cases in group C,30 cases in group E,and 30 cases in group O.The collected tumor tissue specimens were matched with clinical biochemical indexes and tumor markers,and it was found that some indexes were not completely included in the electronic medical record system,so 5 cases in group C,3cases in group E,and 3 cases in group O were excluded,and the final number of included studies was 25 cases in group C,27 cases in group E,and 27 cases in group O,totaling 79 cases.The DNA of the final 79 included subjects was sequenced by16 S r RNA full-length sequencing on the Pac Bio platform,and the sequencing results were used for bioinformatics analysis.Results:1.Comparison of clinical indices among the three groups: the differences in hemoglobin concentration,leukocyte count,platelet count,Neutrophil ratio and Lymphocyte ratio,in the three groups were not statistically significant(P>0.05);while the differences in albumin and CA-125 were significant(P<0.05).2.Comparison of intra-tumor flora in three groups of patients:(1)The diversity of the intra-tumor flora in the three groups of patients was analyzed: the Chao 1 index and Shannon index were slightly higher in groups C and O than in group E,but the differences were not statistically significant.the PCOA results showed significant differences in the intra-tumor flora among individuals in the three groups.In addition,the VENN plot results indicated that the species abundance of the intra-tumor flora was higher in group C than in groups E and O.(2)Differences in intra-tumor flora at the phylum level: there was no significant trend in the relative abundance of the Proteobacteria,Firmicutes,and Actinobacteria at the phylum level,and the differences were not statistically significant.(3)Differences in intra-tumor flora at the genus level: at the genus level,the relative abundance of Pseudomonas was significantly increased in group E compared with groups C and O(P<0.05).The relative abundance of Porphyromonas was significantly increased in group C compared with groups E and O(P<0.05).Compared with groups E and O,group C The relative abundance of Lactobacillus increased significantly compared with groups E and O.3.Effect of age on the distribution of intra-tumoral flora:(1)Effect of age on the distribution of intra-tumor flora in cervical cancer tissues: 25 patients with cervical cancer were divided into two groups according to age,including the low age group(group A,age ≤ 50 years,N=11)and the high age group(group B,age > 50 years,N=14),compared with the low age group(group A),the relative abundance of Bradyrhizobium,Dialister and Lactobacillus differed in the high age group(group B)(P<0.05).(2)Effect of age on the distribution of intratumor flora in endometrial cancer:27 patients with endometrial cancer were divided into two groups according to age,including the low age group(group A,age ≤ 50 years,N=9)and the high age group(group B,age > 50 years,N=18).compared with those in the low age group(group A),the relative abundance of the intra-tumor pathogens Escherichia,Bradyrhizobium and Dialister differed in the high age group(group B)(P<0.05).(3)Effect of age on the distribution of intra-tumor flora in ovarian cancer tissues: 27 ovarian cancer patients were divided into two groups according to their age,including the low age group(group A,age ≤ 50 years,N=13)and the high age group(group B,age > 50 years,N=14).the relative abundance of Staphylococcus in the tumor of patients in the high age group(group B)compared with the lower age group(group A)was different(P<0.05),while the differences of Pseudomonas and Bradyrhizobium were not statistically significant(P>0.05).4.Correlation of intra-tumor flora diversity with clinical biochemical indices and tumor markers:(1)In cervical cancer tissues,the relative abundance of Campylobacter was positively correlated with Leukocyte count(r = 0.402,P = 0.046);the relative abundance of Lactobacillus was negatively correlated with SCCA(r =-0.542,P =0.017).(2)In endometrial cancer tissues,the relative abundance of Finegoldia was not significantly correlated with the lymphocyte ratio(r = 0.373,P = 0.056);the relative abundance of Dialister was negatively correlated with CA-125(r =-0.394,P= 0.042).(3)In ovarian cancer tissues,the relative abundance of Moraxella was negatively correlated with the lymphocyte ratio(r =-0.395,P = 0.042);the relative abundance of Lactobacillus was negatively correlated with CEA(r =-0.518,P =0.016).Conclusions:1.There were significant differences in the diversity and composition of the intra-tumor flora of cervical,endometrial and ovarian cancers,with the highest species abundance in cervical cancer,followed by endometrial cancer and the lowest in ovarian cancer.2.Age is one of the factors affecting the composition of bacteria within the tumor,and the abundance of some pathogenic bacteria in the tumor tissue increases with age.3.Intratumoral flora was associated with some inflammatory indicators and tumor markers,but its specific mechanism of action needs to be confirmed by further studies. |
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