目的：基于基因组高通量测序筛选前列腺癌患者血液中异常表达的miRNAs，并验证其结果。材料与方法：选取在宜春市人民医院泌尿外科的前列腺癌患者、前列腺增生患者及健康人群的血液标本，采用RNA测序（RNA sequencing，RNA-seq）进行测序分析，并运用荧光定量聚合酶链反应（RT-PCR）验证外周血标本中超过10倍以上差异表达基因。通过RT-PCR验证人前列腺癌细胞中miR-429被敲低后的表达水平；分别运用细胞计数试剂盒（CCK-8）实验和EdU细胞染色实验检测细胞的增殖功能。结果：根据测序结果，挑选出在前列腺癌患者外周血中明显高表达的4种miRNAs：miRNA-429、miR-23a、miR-193a和miR-216a-5p，构建相对应的引物，并通过RT-PCR在前列腺癌在患者进行验证发现仅miRNAs-429表达差异较为明显。构建的miRNAs-429质粒可显著上调的前列腺癌细胞增殖能力，并降低细胞凋亡。结论：miRNAs-429的异常表达对前列腺癌细胞增殖和细胞凋亡均有影响，这些结果表明miRNAs-429在前列腺癌的发生、发展中可能发挥重要的作用。

关键字：前列腺癌；miRNA；miR-429；细胞增殖；测序

Abstract：

Objective: To screen the abnormal expression of miRNAs in the blood of prostate cancer patients based on high-throughput genome sequencing and verify the results. Materials and Methods: Blood samples from prostate cancer patients, prostate hyperplasia patients and healthy people in the Department of Urology of Yichun People's Hospital were selected for analysis through RNA sequencing (RNA-SEQ). Fluorescence quantitative polymerase chain reaction (RT-PCR) was used to verify more than 10 times differentially expressed genes in peripheral blood samples. The expression level of miR-429 in human prostate cancer cells after knockdown was verified by RT-PCR. Cell counting kit (CCK-8) assay and EdU cell staining assay were used to detect cell proliferation. Results: According to the sequencing results, 4 miRNAs with high expression in peripheral blood of patients with prostate cancer were selected: The corresponding primers were constructed for miRNA-429, miR-23a, miR-193a and miR-216a-5p, and it was found that only miRNAs-429 expression was significantly different in patients with prostate cancer by RT-PCR. The constructed miRNAs-429 plasmid can significantly up-regulate the proliferation of prostate cancer cells and reduce cell apoptosis. Conclusion: The abnormal expression of miRNAs-429 may affect the proliferation and apoptosis of prostate cancer cells. These results suggest that miRNAs-429 may play an important role in the occurrence and development of prostate cancer.

Keywords: Prostate cancer; miRNA; miR-429; Cell proliferation; sequencing

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