【摘要】：目的：MicroRNAs(miRNAs)是一類長約20～22個核苷酸的非編碼單鏈RNA分子,可通過與其相關的mRNA分子3’端非編碼區(qū)(untranslated region, UR)互補配對,參與基因表達的轉錄后調控。大量證據證明miRNA在腫瘤的發(fā)展,分化,凋亡及增殖過程中發(fā)揮著關鍵作用。在前列腺癌中,miR-154的表達及具體功能目前研究甚少,本課題擬驗證miR-154在前列腺癌中的表達趨勢及研究其對前列腺癌細胞生物學功能的影響。 材料和方法：采用qRT-PCR方法檢測27例前列腺癌組織和9例前列腺正常組織的樣本中miR-154的表達水平。運用細胞增殖實驗、集落形成實驗、細胞周期實驗及Western blot分析評估m(xù)iR-154和CCND2的表達變化對前列腺細胞系PC-3和DU145的影響。通過雙熒光素酶報告基因實驗確定miR-154是否通過CCND2的3'-UTR區(qū)域調控CCND2。 結果：與癌旁前列腺正常組織相比,miR-154在前列腺癌組織中的表達水平明顯下調。而與Gleason評分、T分級相關無明顯相關性。體外實驗中(CCK-8、集落形成實驗、細胞周期分析)上調miR-154的表達水平和下調CCND2表達水平可以明顯抑制前列腺癌細胞的增殖能力。這種關系表明,在腫瘤的發(fā)生發(fā)展過程中miR-154與CCND2起協(xié)同作用。雙熒比素酶報告實驗證明CCND2為miR-154直接調控的靶基因。轉染miR-154mimics后72小時后,細胞內CCND2蛋白表達水平顯著降低。 結論：在前列腺癌中miR-154可作為一種腫瘤抑制因子,直接在轉錄后水平調控CCND2的表達。上調miR-154或下調CCND2的表達水平均可影響前列腺癌細胞的遷移和侵襲能力。miR-154有希望成為診斷和治療前列腺癌的分子標記和靶點。
[Abstract]:Objective: microRNAs (miRNAs) are a class of non-coding single-stranded RNAs with a length of about 20 ~ 22 nucleotides. They may be involved in the posttranscriptional regulation of gene expression through complementary pairing of the 3 '-terminal noncoding region (untranslated region, UR of their related mRNAs. A great deal of evidence shows that miRNA plays a key role in tumor development, differentiation, apoptosis and proliferation. The expression and specific function of miR-154 in prostate cancer are rarely studied. This study aims to verify the expression trend of miR-154 in prostate cancer and its effect on the biological function of prostate cancer cells. Materials and methods: the expression of miR-154 was detected by qRT-PCR in 27 prostate cancer tissues and 9 normal prostate tissues. The effects of miR-154 and CCND2 expression on prostate cell line PC-3 and DU145 were evaluated by cell proliferation assay, colony formation assay, cell cycle assay and Western blot analysis. Whether miR-154 regulates CCND2 through CCND2 3- UTR region is determined by double luciferase reporter gene experiment. Results: the expression of miR-154 in prostate cancer tissues was significantly down-regulated compared with the adjacent normal prostate tissues. There was no significant correlation between Gleason score and T grade. In vitro (CCK-8, colony forming assay, cell cycle analysis), upregulation of miR-154 expression and down-regulation of CCND2 expression could significantly inhibit the proliferation of prostate cancer cells. This relationship suggests that miR-154 and CCND2 play a synergistic role in tumorigenesis and development. The double fluorinase report showed that CCND2 was the target gene directly regulated by miR-154. After 72 hours of miR-154 mimics transfection, the expression of CCND2 protein decreased significantly. Conclusion: miR-154 may act as a tumor suppressor in prostate cancer and regulate the expression of CCND2 directly at posttranscriptional level. Upregulation of miR-154 or down-regulation of CCND2 expression may affect the migration and invasion of prostate cancer cells. MiR-154 may become a molecular marker and target for the diagnosis and treatment of prostate cancer.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R737.25

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