miR-154通过调控CCND2影响前列腺癌细胞的增殖能力
发布时间：2018-07-21 20:32
【摘要】：目的：MicroRNAs(miRNAs)是一类长约20～22个核苷酸的非编码单链RNA分子,可通过与其相关的mRNA分子3’端非编码区(untranslated region, UR)互补配对,参与基因表达的转录后调控。大量证据证明miRNA在肿瘤的发展,分化,凋亡及增殖过程中发挥着关键作用。在前列腺癌中,miR-154的表达及具体功能目前研究甚少,本课题拟验证miR-154在前列腺癌中的表达趋势及研究其对前列腺癌细胞生物学功能的影响。 材料和方法：采用qRT-PCR方法检测27例前列腺癌组织和9例前列腺正常组织的样本中miR-154的表达水平。运用细胞增殖实验、集落形成实验、细胞周期实验及Western blot分析评估miR-154和CCND2的表达变化对前列腺细胞系PC-3和DU145的影响。通过双荧光素酶报告基因实验确定miR-154是否通过CCND2的3'-UTR区域调控CCND2。 结果：与癌旁前列腺正常组织相比,miR-154在前列腺癌组织中的表达水平明显下调。而与Gleason评分、T分级相关无明显相关性。体外实验中(CCK-8、集落形成实验、细胞周期分析)上调miR-154的表达水平和下调CCND2表达水平可以明显抑制前列腺癌细胞的增殖能力。这种关系表明,在肿瘤的发生发展过程中miR-154与CCND2起协同作用。双荧比素酶报告实验证明CCND2为miR-154直接调控的靶基因。转染miR-154mimics后72小时后,细胞内CCND2蛋白表达水平显著降低。 结论：在前列腺癌中miR-154可作为一种肿瘤抑制因子,直接在转录后水平调控CCND2的表达。上调miR-154或下调CCND2的表达水平均可影响前列腺癌细胞的迁移和侵袭能力。miR-154有希望成为诊断和治疗前列腺癌的分子标记和靶点。
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25
[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.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25
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