肠道菌群与乳腺癌患者预后不良的关系

Relationship between intestinal flora and poor prognosis of breast cancer patients

目的分析肠道菌群与乳腺癌患者预后不良的关系。方法选取2020年1月至2022年12月深圳市第二人民医院收治的225例乳腺癌患者(疾病组)和225例健康对照人群(健康组)作为研究对象。采用聚合酶链反应(PCR)及高通量测序测定两组粪便标本的肠道菌群。统计并比较两组丰富度(Chao指数)、多样性(Shannon指数)及属水平相对丰度。疾病组患者实施常规抗肿瘤治疗,定期随访,根据预后结果分为预后不良组和预后良好组,统计预后不良发生率。比较预后不良组与预后良好组一般资料及肠道菌群Chao指数、Shannon指数、属水平相对丰度;采用受试者工作特征(ROC)曲线分析差异有统计学意义的肠道菌群指标对乳腺癌患者预后不良的预测价值;采用Kaplan-Meier生存曲线分析筛选变量,并采用多因素Cox回归分析探讨乳腺癌患者预后不良的影响因素。结果疾病组Chao指数、Shannon指数及双歧杆菌属、肠球菌属、乳杆菌属、链球菌属、普氏菌属相对丰度均低于健康组(P<0.05),瘤胃球菌属、布劳特菌属相对丰度均高于健康组(P<0.05)。乳腺癌患者中预后不良发生率为30.56%(66/216)。预后不良组Ⅲ~Ⅳ期、三阴性乳腺癌、未/低分化比例及肠道菌群瘤胃球菌属、布劳特菌属相对丰度均高于预后良好组(P<0.05),Chao指数、Shannon指数及双歧杆菌属、乳杆菌属、普氏菌属相对丰度均低于预后良好组(P<0.05)。ROC曲线分析结果显示:Chao指数、Shannon指数及乳杆菌属、普氏菌属、双歧杆菌属、瘤胃球菌属、布劳特菌属相对丰度预测乳腺癌患者预后不良的cut-off值分别为44.01、0.69、13.78%、3.97%、6.77%、7.92%、9.74%,曲线下面积(AUC)分别为0.893(95%CI:0.844~0.931)、0.949(95%CI:0.911~0.974)、0.862(95%CI:0.809~0.905)、0.852(95%CI:0.797~0.897)、0.782(95%CI:0.721~0.835)、0.744(95%CI:0.680~0.800)、0.721(95%CI:0.656~0.780)。经Kaplan-Meier生存曲线分析,临床分期、分化程度及双歧杆菌属、瘤胃球菌属相对丰度的Kaplan-Meier生存曲线无交叉,满足比例风险假定,故作为自变量,进行乳腺癌患者预后不良影响因素的多因素Cox回归分析,结果显示:Ⅲ~Ⅳ期、未/低分化、瘤胃球菌属相对丰度升高均是乳腺癌患者预后不良的危险因素(P<0.05),双歧杆菌属相对丰度升高是乳腺癌患者预后不良的保护因素(P<0.05)。结论乳腺癌患者肠道菌群丰富度及多样性下降,双歧杆菌属、肠球菌属、乳杆菌属、链球菌属、普氏菌属相对丰度降低,而瘤胃球菌属、布劳特菌相对丰度升高。瘤胃球菌属相对丰度升高、Ⅲ~Ⅳ期、三阴性乳腺癌、未/低分化均是乳腺癌患者预后不良的危险因素,双歧杆菌属相对丰度升高是乳腺癌患者预后不良的保护因素。

Objective To analyze the relationship between intestinal flora and poor prognosis of breast cancer patients.Methods Two hundred and twenty-five patients with breast cancer(disease group)and 225 healthy control subjects(health group)admitted to Shenzhen Municipal Second People′s Hospital from January 2020 to December 2022 were selected as the study subjects.The polymerase chain reaction(PCR)and high-throughput sequencing were used to determine the intestinal flora of fecal specimens in the two groups.The richness(Chao index),diversity(Shannon index)and relative abundance at the genus level were statistically compared between two groups.All patients in the disease group were treated with conventional anti-tumor therapy and followed up regularly.They were divided into the poor prognosis group and good prognosis group.The incidence rate of poor prognosis was calculated;the general data,Chao index,Shannon index,and relative abundance of intestinal flora at the genus level were compared between the poor prognosis group and good prognosis group;the predictive value of statistically different indicators of intestinal flora for poor prognosis of breast cancer patients was analyzed by the receiver operating characteristic(ROC)curve;the Kaplan-Meier survival curve analysis was used to screen the variables,and the multivariate Cox regression analysis was used to analyze the influencing factors of poor prognosis in breast cancer patients.Results The Chao index,Shannon index and relative abundances of Bifidobacterium,Enterococcus,Lactobacillus,Streptococcus,and Prevotella in the disease group all were lower than those the healthy group(P<0.05),while the relative abundances of Ruminococcus and Brucella were higher than those in the healthy group(P<0.05).The incidence rate of poor prognosis in breast cancer patients was 30.56%(66/216).The proportions of the stageⅢ-Ⅳ,triple negative breast cancer and undifferential/poorl differentiation,and the relative abundances of the intestinal flora Ruminococcus and Brucella in the poor prognosis group all were higher than those in the good prognosis group(P<0.05);the Chao index,Shannon index,and the relative abundances of Bifidobacterium,Lactobacillus and Proctor were lower than those in the good prognosis group(P<0.05).The ROC curve analysis results showed that the cut-off values of Chao index,Shannon index,and the relative abundances of Lactobacillus,Prevotella,Bifidobacterium,Ruminococcus,and Brucella for predicting poor prognosis of breast cancer patients were 44.01,0.69,13.78%,3.97%,6.77%,7.92%,and 9.74%respectively,and the areas under the curve(AUC)were 0.893(95%CI:0.844-0.931),0.949(95%CI:0.911-0.974),0.862(95%CI:0.809-0.905),0.852(95%CI:0.797-0.897),and 0.782(95%CI:0.721-0.835),0.744(95%CI:0.680-0.800),0.721(95%CI:0.656-0.780)respectively.The Kaplan-Meier survival curve analysis showed that the clinical stage,degree of differentiation,no cross of Kaplan-Meier survival curve of the relative abundances of Bifidobacterium and Ruminococcus,which met the proportional risk assumption,therefore served as the independent variables,the Cox regression analysis of the influencing factors of the poor prognosis in breast cancer patients was conducted.The results showed that the stageⅢ-Ⅳ,undifferentiation/low differentiation and Ruminococcus relative abundance increase all were the risk factors for poor prognosis in breast cancer patients(P<0.05),and the Bifidobacterium relative abundance increase was the protective factor for poor prognosis in breast cancer patients(P<0.05).Conclusion The richness and diversity of intestinal flora in the patients with breast cancer are decreased,and the relative abundance of Bifidobacterium,Enterococcus,Lactobacillus,Streptococcus and Prevotella are decreased,while the relative abundance of Rumen Coccus and Brucella are increased.The Ruminococcus genus relative abundance increase,stageⅢ-Ⅳ,triple negative breast cancer and undifferentiation/poor differentiation are the risk factors for poor prognosis in breast cancer patients.The Bifidobacterium relative abundance increase is a protective factor for poor prognosis in breast cancer patients.