The Expression of Hepatocyte Growth Factor (HGF) and Its Receptor (C-MET) in Human Benign Prostatic Hyperplasia and Prostate Cancer

肝细胞生长因子（HGF）及其受体（C-MET）在人良性前列腺增生和前列腺癌中的表达

• 批准号: 07671712
• 负责人: MORI Hirotaka
• 金额: $ 0.13万
• 依托单位: Fukui Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671712/
• 关键词: prostate; hepatocyte growth factor; C-MET; 前立腺肥大症; 前立腺癌

It is demonstrated that prostatic growth is regulated by several growth factors in addition to androgen. We have been investigating the expression of some of the growth factors in human prostatic tissue. We analyzed the expression of HGF and C-MET in the prostate.The human prostate in liquid nitrogen is stony-hard and is very difficult to be homogenized before preparing RNA.We found a relatively easy and reliable method for the preparation of intact RNA from hard frozen tissue of the human prostate. Frozen tissue was freeze-dried before cutting into small pieces and homogenizedin TRIzol reagent (GIBCOBRL). The conventional method was to crush tissue specimens to small pieces within 1mm in diameter in liquid nitrogen at 15000 rpm for 1 hour with a Waring blender before homogenization in TRIzol. The quality and quantity of RNA between two methods was compared in Northern blotting and RT-PCR.Freeze-drying method was as efficient as the conventional method. Freeze-dried tissue could be stored at 4ﾟC for one month without degradation of RNA.After obtaining RNA from human prostatic tissue specimens (normal, benign prostatic hyperplasia, prostate cancer), the expressions of HGF and C-MET were analyzed with competitive RT-PCR.When small number of samples were examined, C-MET expression was elevated in the normal prostate. There was no difference in HGF mRNA quantity among three groups. We are now investigating quantity and localization of both proteins using Western blotting and immunohistochemistry.

它表明，前列腺的生长是由几个生长因子，除了雄激素。我们一直在研究一些生长因子在人前列腺组织中的表达。我们分析了HGF和C-MET在前列腺组织中的表达情况，由于人前列腺在液氮中非常坚硬，在制备RNA之前很难进行匀浆，因此我们找到了一种相对简单可靠的从人前列腺组织中制备完整RNA的方法。将冷冻组织冷冻干燥，然后切成小块并在TRIzol试剂（GIBCOBRL）中均质化。常规方法是将组织样本在液氮中以15000 rpm粉碎1小时，然后在TRIzol中均质化。通过北方杂交和RT-PCR比较了两种方法提取RNA的质量和数量，结果表明，冷冻干燥法提取RNA的效率与常规方法相当。从人前列腺组织（正常、良性前列腺增生、前列腺癌）中提取RNA，采用竞争性RT-PCR方法检测HGF和C-MET的表达，在少量标本中，正常前列腺组织中C-MET的表达升高。三组间HGF mRNA表达量无显著性差异。我们现在正在调查的数量和定位这两种蛋白质使用Western印迹和免疫组织化学。

项目成果

Harutoshi Tsuka, Hirotaka Mori, Kenichiro Okada, and Shigeru Matsukawa: Analytical Biochemistry. (accepted for publication). (1997)

Harutoshi Tsuka、Hirotaka Mori、Kenichiro Okada 和 Shigeru Matsukawa：分析生物化学。

Harutoshi Tsuka: "Utilization of freeze-drying method in the preparation of biologically active, intact RNA from hard frozen tissues of human prostate" Analytical Biochemistry.(accepted). (1997)

Harutoshi Tsuka：“利用冷冻干燥方法从人类前列腺硬冷冻组织中制备具有生物活性的完整 RNA”《分析生物化学》（已接受）。

Harutoshi Tsuka: "Utilization of freeze-drying method in the preparation of biologically active,intact RNA from hard frozen tissues of human prostate" Analytical Biochemistry,accepted. (1997)

Harutoshi Tsuka：“利用冷冻干燥方法从人类前列腺的硬冷冻组织中制备具有生物活性的完整 RNA”《分析生物化学》，已接受。