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在前列腺癌小鼠模型中解剖单个前列腺叶以获取高质量RNA。

Dissection of Individual Prostate Lobes in Mouse Models of Prostate Cancer to Obtain High Quality RNA.

作者信息

Zingiryan Areg, Farina Nicholas H, Finstad Kristiaan H, Stein Janet L, Lian Jane B, Stein Gary S

机构信息

Department of Biochemistry, University of Vermont Cancer Center, University of Vermont College of Medicine, Burlington, Vermont.

出版信息

J Cell Physiol. 2017 Jan;232(1):14-8. doi: 10.1002/jcp.25384. Epub 2016 Apr 1.

Abstract

Genetically engineered mouse models of prostate cancer allow for study of disease progression from localized tumor formation through distal metastasis. The anatomy of the mouse prostate differs dramatically from the human prostate, being composed of four lobe pairs (anterior, dorsal, lateral, and ventral), making the identification and dissection technically challenging. Although the entire murine prostate and surrounding tissue, including urethra, bladder, seminal vesicles, and associated adipose tissue, can be quickly dissected for en bloc analysis, it is necessary to isolate individual prostate lobes for gene expression studies elucidating the molecular mechanisms of prostate cancer. The procedure as described here includes full color images, allowing the researcher to appreciate the unique prostate morphology and tissue manipulation required to harvest individual prostate lobes. Along with removing all extraneous tissue, the procedure allows for direct comparison of the different prostate lobes by established downstream techniques. Importantly, high quality RNA required for next-generation gene expression analysis can only consistently be obtained from ventral and lateral lobes. Finally, preclinical studies using prostate targeted therapies can be monitored specifically in individual prostate lobes for histological and gene expression studies. J. Cell. Physiol. 232: 14-18, 2017. © 2016 Wiley Periodicals, Inc.

摘要

前列腺癌的基因工程小鼠模型有助于研究疾病从局部肿瘤形成到远处转移的进展过程。小鼠前列腺的解剖结构与人类前列腺有很大差异,它由四对叶(前叶、背叶、侧叶和腹叶)组成,这使得识别和解剖在技术上具有挑战性。尽管整个小鼠前列腺及周围组织,包括尿道、膀胱、精囊和相关脂肪组织,可以快速进行整块解剖用于整体分析,但为了阐明前列腺癌的分子机制进行基因表达研究时,有必要分离出各个前列腺叶。这里描述的操作步骤配有全彩图像,使研究人员能够了解获取各个前列腺叶所需的独特前列腺形态和组织操作。该操作在去除所有多余组织的同时,还能通过既定的下游技术对不同的前列腺叶进行直接比较。重要的是,下一代基因表达分析所需的高质量RNA通常只能从腹叶和侧叶中稳定获取。最后,使用前列腺靶向疗法的临床前研究可以在各个前列腺叶中进行专门监测,用于组织学和基因表达研究。《细胞生理学杂志》232: 14 - 18, 2017。© 2016威利期刊公司。

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