不同临床分期前列腺癌患者精浆microRNA谱鉴定

Cell-free microRNA (miRNA) is found to be actively secreted by living cells and passively released via necrotic cells, they can provide the miRNA expression information of derived cell and be used for the diagnosis, outcome prediction and treatment monitoring of clinical disease. Our previous result showed that the concentration of cell-free RNA was high in prostatic fluid, and their miRNA was stable in room temperature, which is accordant with our reports about the seminal plasma. Considering the prostatic fluid accounts for about one third of seminal plasma, then we suppose that seminal plasma may be a noninvasive way of reflecting the prostate-specific miRNA expression information . Moreover, the miRNA of seminal plasma has the potential to be the ideal substitute of prostate biopsy for studying the expression level of prostate-specific miRNA in prostate cancer (PCa). The project will research the miRNA profiles in the seminal plasma of PCa patients with different clinical stages, then screening and validate the prostate-specific miRNAs of different clinical stages in small PCa patients by qRT-PCR. These verified miRNAs can provide the noninvasive candidate markers for the early diagnosis, outcome prediction and treatment monitoring of PCa. At the same time, we also confirm some prostate-specific miRNAs of clinical stages which are highly consistent with the clinical stages in some PCa patients. Therefore, these miRNAs can be used for the noninvasive diagnosis of clinical stages in PCa patients.

游离microRNA (miRNA)是活细胞主动分泌或死亡细胞被动释放miRNA，可提供所来源细胞miRNA表达信息而被用于疾病诊断、预后评估和疗效监测。前期研究结果显示，前列腺液游离总RNA浓度高且其miRNA体外稳定，与我们精浆报道的结果一致。而前列腺液约占精浆1/3，推测精浆可能成为反映前列腺特异性miRNA表达信息的无创性途径，并可望成为研究前列腺癌 （PCa）患者前列腺组织特异性miRNA表达水平的理想前列腺穿刺替代物。本课题研究不同临床分期PCa患者精浆miRNA谱，并筛选和验证PCa临床分期特异性miRNAs，为PCa早期诊断、预后评估和疗效监测提供无创候选标记物。并在不同PCa个体验证出部分与PCa临床分期符合率高的特异性miRNAs，为PCa临床分期无创诊断提供标记物。

前列腺癌（prostate cancer, PCa）治疗及预后评估取决于其准确的临床分期，目前PCa的临床分期主要依靠前列腺组织穿刺及影像学检查，而前列腺组织穿刺为有创性检查且漏诊率达(30-50)％，影像学检查无法发现PCa微小的原发及转移病灶，这些因素限制了PCa临床分期的准确性。考虑到前列腺液约占精液体积的1/3，推测精浆中前列腺液来源的细胞游离miRNA能全面反映前列腺组织miRNA表达信息，从而代替前列腺组织miRNA用于PCa临床分期的无创诊断。为了发现精浆中PCa不同临床分期特异性miRNAs，我们对前列腺正常、前列腺增生、原位PCa、局部进展期PCa和转移性PCa患者精浆混合总RNA进行solexa测序，在原位PCa、局部进展期PCa及转移性PCa精浆中分别发现了13个、9个、15个高表达的miRNAs及10个、6个、11个低表达的miRNAs。并通过qRT-PCR方法对这些特异性miRNA进行验证，发现有70.31%(45/64) miRNA与solexa测序结果一致。根据精浆中前列腺癌不同临床分期特异性miRNAs靶基因功能的分析，分别选择了在原位PCa、局部进展期PCa及转移性PCa精浆中高表达的5个、4个、5个miRNAs及低表达的5个、3个、5个miRNAs，对前列腺正常 (15例)、前列腺增生 (15例)、原位PCa (14例)、局部进展期PCa (12例)及远处转移PCa (15例)受试者的精浆表达水平进行了检测，分别在原位PCa、局部进展期PCa、远处转移PCa发现了5个、3个、6个特异性高的miRNAs，这些miRNAs有望用于PCa患者的早期诊断、临床分期、预后评估、疗效监测等。