高级别浆液性卵巢癌组微生物失衡初步研究

发布时间：2018-05-07 00:45

本文选题：微生物 + 卵巢癌　；参考：《华中科技大学》2016年博士论文

【摘要】：目的：当前,微生物与疾病,尤其是与肿瘤的发生发展越来越受到科研工作者的重视和研究。卵巢癌的起源目前仍没有得到一致的结论,而且卵巢癌组织中的微生物组分目前几乎没有研究报道。本研究拟对高级别浆液性卵巢癌组织和正常输卵管组织中微生物群落组成和多样性进行研究。方法：收集来自武汉同济医院妇产科2013年10月到2015年10月71例新鲜高级别浆液性卵巢癌组织和66例新鲜正常输卵管伞端组织标本,利用16S rRNA高通量测序的方法对其中25例卵巢癌组织和25例正常输卵管组织进行测序。另外,利用实时荧光定量PCR方法46例新鲜卵巢癌组织和41例新鲜正常输卵管组织的部分差异微生物进行独立样本的验证以及受试者工作特征曲线分析。最后,通过人抗细菌反应的高通量PCR芯片检测抗细菌反应的相关基因和信号通路的表达情况。结果：(一)对新鲜组织样本16S rRNA高通量测序显示了人体卵巢癌组织和输卵管组织中微生物的组成的变化：(二)基于可操作的分类学单元(OTUs)卵巢癌组织中微生物数量相对正常输卵管组织微生物数量减少；(三)通过LEfSe联合统计学分析,采用实时荧光定量PCR证实部分显著差异微生物的丰富度和16S高通量测序结果相吻合。在门的水平上,变形菌门Proteobacteria的丰度在卵巢癌组织中相对正常输卵管组织增加,而厚壁菌门Firmicutes的丰度在卵巢癌组织中相对正常输卵管组织中下降,具有统计学意义,且与16S测序数据一致；在属的水平上,不动杆菌属Acinetobacter的丰度在卵巢癌组织中较高,具有统计学意义,乳球菌属Lactococcus的丰度在卵巢癌组织中减少,与16S测序结果相符合；在种的水平上,鲁菲不动杆菌种A cinetobacter_lwoffii的丰度在卵巢癌组织中增加,具有统计学意义且与16S测序结果一致,而Lactococcus_psicium在卵巢癌组织中明显下降,具有明显的统计学意义,与16S测序结果吻合；(四)基于人抗细菌反应的PCR芯片显示,相对于正常输卵管组织中的微生物的基因表达,卵巢癌组织中人抗细菌反应的基因表达大部分下调。结论：首次证实卵巢癌组织相对正常输卵管组织中微生物数量降低,呈现失衡状态,我们假设微生物失衡可能影响卵巢癌的发生和发展,其可能的机制是造成局部炎症反应,进而影响肿瘤局部免疫微环境。我们的研究提供了部分卵巢癌和正常输卵管组织中微生物的一个数据集,为卵巢癌的发生,进展,早期诊断和治疗提供了潜在的价值。
[Abstract]:Objective: at present, microorganism and disease, especially the occurrence and development of tumor, are paid more and more attention by researchers. The origin of ovarian cancer has not been unanimous, and the microbial components in ovarian cancer tissues have not been reported. This study was designed to study the composition and diversity of microbial communities in high grade serous ovarian cancer tissues and normal oviduct tissues. Methods: from October 2013 to October 2015, 71 samples of fresh high-grade serous ovarian cancer and 66 cases of fresh normal fallopian tubes were collected from gynecology and obstetrics department of Tongji Hospital, Wuhan. High throughput 16s rRNA sequencing was performed in 25 ovarian cancer tissues and 25 normal fallopian tube tissues. In addition, 46 fresh ovarian cancer tissues and 41 fresh normal fallopian tube tissues were tested by real-time fluorescence quantitative PCR method for the verification of partial differential microbes and the analysis of the operating characteristic curves. Finally, high throughput PCR microarray was used to detect the expression of genes and signaling pathways related to antimicrobial reactions. Results: (1) High throughput sequencing of 16s rRNA from fresh tissue samples showed changes in the composition of microbes in human ovarian cancer tissues and oviduct tissues. (2) the number of microbes in ovarian cancer tissues was based on an operable taxonomic unit. The amount of microorganism in the normal oviduct tissue is reduced (3) through LEfSe combined with statistical analysis, The results of 16s high throughput sequencing showed that the abundance of some significantly different microbes was consistent with the results of 16s high throughput sequencing by real-time fluorescence quantitative PCR. At the gate level, the abundance of Proteobacteria in ovarian cancer was higher than that in normal fallopian tube, while the abundance of Firmicutes was decreased in ovarian cancer, which was statistically significant. In accordance with 16s sequencing data, the abundance of Acinetobacter was higher in ovarian cancer tissues, and the abundance of Lactococcus Lactococcus decreased in ovarian cancer tissues, which was consistent with the results of 16s sequencing. At the seed level, the abundance of A cinetobacter_lwoffii of Acinetobacter rufii increased in ovarian cancer tissues, which was statistically significant and consistent with 16s sequencing results, while Lactococcus_psicium decreased significantly in ovarian cancer tissues. In accordance with 16s sequencing results (4) PCR microarray based on human anti-bacterial reaction showed that the gene expression of human anti-bacterial reaction was mostly down-regulated in ovarian cancer compared with that of microorganism in normal oviduct tissue. Conclusion: it is the first time to confirm that the number of microbes in ovarian cancer tissue is lower than that in normal oviduct tissue. We hypothesized that the microbial imbalance may affect the occurrence and development of ovarian cancer, and the possible mechanism is to cause local inflammatory reaction. And then affect the tumor local immune microenvironment. Our study provides a data set of microorganisms in some ovarian cancer and normal fallopian tube tissues, providing potential value for the occurrence, progression, early diagnosis and treatment of ovarian cancer.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R737.31

【相似文献】