非编码RNA在乳腺癌Tamoxifen抗性和男性不育发生中的功能研究

发布时间：2018-05-23 06:42

本文选题：miR-320a + ARPP-19　；参考：《中国科学技术大学》2015年博士论文

【摘要】：Tamoxifen作为治疗雌激素受体阳性乳腺癌病人的一种主要的辅助性药物。但是,在用Tamoxifen治疗过程中,病人容易产生tamoxifen抗性,有的是先天性的,有的是在治疗过程中产生的。为了研究miR-320a在产生tamoxifen抗性过程发挥的作用,我们通过让乳腺癌细胞系MCF-7和T47D细胞长期暴露在1μM的tamoxifen中,建立了tamoxifen抗性的细胞系MCF-7-TamR和T47D-TamR细胞。Real-time PCR实验发现,与母细胞相比,miR-320a在MCF-7-TamR和T47D-TamR细胞中表达明显降低。过表达miR-320a可以使MCF-7-TamR和T47D-TamR细胞对tamoxifen重新产生敏感性,miR-320a行使这一功能是通过抑制其靶基因ARPP-19和ERRγ及其下游基因c-Myc和Cyclin D1的表达。另外,孕酮(P4)通过c-Myc可以促进miR-320s滚达,雌二醇(E2)可以同过c-Myc抑制miR-320a的表达。这些结果表明,可以通过恢复miR-320a的表达或者敲除APPP-19和ERRy的表达来消除tamoxifen抗性,使得乳腺癌病人重新对tamoxife产生敏感性。基因组普遍转录的长非编码RNA(lncRNAs)作为主要调节子参与各个生物发生过程。但是,有关长非编码RNA参与人类男性精子发生过程却知道很少。利用微阵列技术,我们发现与正常男性睾丸相比,在成熟停滞的不育男性睾丸组织中,有757个下调的长非编码RNA,475个上调的长非编码RNA；在寡精症的不育男性睾丸组织中,有2370个下调的长非编码RNA,163个上调的长非编码RNA。利用real-time PCR (qRT-PCR)技术对选择的lncRNAs(包括：HOTTIP, imsrna320和NLC1-C)进行分析,验证了这一结果。非常有趣的是,与正常组织相比,NLC1-C,也称做基因间长非蛋白编码RNA162(LINC00162),在成熟停滞的睾丸组织精原细胞和初级精母细胞质中表达降低,积累在细胞核中。积累在细胞核中的NLC1-C通过与Nucleolin蛋白结合抑制miR-320a/383转录和促进睾丸癌细胞增殖。在我们的研究中,我们发现了一种新的长非编码RNA与蛋白结合通过调控miRNA的转录进而调控精子发生的机制。
[Abstract]:Tamoxifen is a major adjuvant drug for the treatment of estrogen receptor positive breast cancer patients. However, in the course of Tamoxifen treatment, patients are prone to produce tamoxifen resistance, some are congenital and some are produced during the treatment process. In order to study the role of miR-320a in the production of tamoxifen resistance, we are involved. The MCF-7 and T47D cells of the breast cancer cell lines were exposed to the tamoxifen of 1 mu M for a long time. The tamoxifen resistant cell line MCF-7-TamR and T47D-TamR cell.Real-time PCR experiments were established. Compared with the mother cell, the expression of miR-320a in MCF-7-TamR and T47D-TamR cells decreased significantly. The cell reproduces sensitivity to tamoxifen. MiR-320a exercises this function by inhibiting the expression of its target gene ARPP-19 and ERR gamma and its downstream gene c-Myc and Cyclin D1. In addition, progesterone (P4) can promote miR-320s to roll through c-Myc, and estradiol (E2) can be expressed with over c-Myc inhibition. The expression of -320a or knockout of APPP-19 and ERRy expression can eliminate tamoxifen resistance and make breast cancer patients re sensitize to tamoxife.
Long non coded RNA (lncRNAs), which is the main regulator of the genome, participates in the process of biogenesis as the main regulator. However, there is little knowledge about the involvement of long non coded RNA in human spermatogenesis. By microarray technology, we found that there are 757 in mature sterility male testicular tissues compared with normal male testicles. Down regulated long non coding RNA, 475 up-regulated long noncoding RNA; in oligospermia male infertility male testicular tissues, there were 2370 down-regulated long non coded RNA and 163 up - regulated long non coded RNA. used real-time PCR (qRT-PCR) technique to analyze selected lncRNAs (including HOTTIP, imsrna320 and NLC1-C). The results were verified. Interestingly, compared with normal tissue, NLC1-C, also known as a long non protein encoded RNA162 (LINC00162), decreases in mature spermatogonia and primary spermatogenic cytoplasm, accumulates in the nucleus. The accumulation of NLC1-C in the nucleus by binding to the Nucleolin protein inhibits miR-320a/383 transcription and promotes testicles. Cancer cell proliferation. In our study, we have found a new mechanism that combines long noncoding RNA with protein to regulate the transcription of miRNA and then regulate spermatogenesis.
【学位授予单位】：中国科学技术大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.9

【共引文献】