转移相关蛋白ATP5B的鉴定及其在PC3M细胞中的表达及定位
发布时间：2018-08-09 09:09
【摘要】：侵袭和转移是前列腺癌患者的重要死因。目前人类对前列腺癌的认识水平有限，常规的治疗手段尚无法从根本上清除恶性肿瘤，因此明确前列腺癌侵袭、转移发生的关键机制，寻找相应的阻断途径，建立起一套行之有效的治疗手段就显得尤为重要。 前期研究发现只与前列腺癌高转移细胞系PC3M特异性结合的短肽B04具有抑制PC3M细胞侵袭和转移的能力，因此我们认为其在PC3M细胞上的受体可能与前列腺癌的转移有关。 为了获取特异性短肽B04的受体，本研究采用western blot、考马斯亮蓝染色、银染色等方法初步定位特异性短肽B04受体的表达，继而采用亲和层析和质谱鉴定等方法鉴定受体蛋白的身份信息，，并通过免疫荧光细胞化学染色的方法对受体蛋白的亚细胞定位进行分析。结果证实短肽B04的受体蛋白为PC3M细胞的膜蛋白，成功地鉴定出ATP5B为特异性短肽B04的受体蛋白之一；证明ATP5B表达于前列腺癌高转移细胞系PC3M的线粒体和细胞膜上，发现ATP5B为前列腺癌转移相关蛋白；ATP5B表达于细胞膜上可能是其发挥转移相关功能的关键。
[Abstract]:Invasion and metastasis are important causes of death in patients with prostate cancer. At present, the level of human understanding of prostate cancer is limited, conventional treatment methods can not fundamentally remove malignant tumors, so the key mechanism of prostate cancer invasion and metastasis is identified, and the corresponding blocking pathway is found. It is particularly important to establish a set of effective treatment methods. Previous studies have shown that the short peptide B04, which specifically binds to high metastatic prostate cancer cell line PC3M, can inhibit the invasion and metastasis of PC3M cells. Therefore, we believe that its receptor on PC3M cells may be related to the metastasis of prostate cancer. In order to obtain the receptor of specific short peptide B04, western blot, Coomassie brilliant blue staining and silver staining were used to preliminarily localize the expression of specific short peptide B04 receptor. Then the identity information of the receptor protein was identified by affinity chromatography and mass spectrometry, and the subcellular localization of the receptor protein was analyzed by immunofluorescence cytochemical staining. The results showed that the receptor protein of short peptide B04 was the membrane protein of PC3M cells, and that ATP5B was one of the receptor proteins of short peptide B04, and that ATP5B was expressed in mitochondria and cell membrane of high metastatic prostate cancer cell line PC3M. It was found that the expression of ATP5B in the cell membrane of the metastasis associated protein of prostate cancer may be the key to its metastasis-related function.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25
本文编号：2173614
[Abstract]:Invasion and metastasis are important causes of death in patients with prostate cancer. At present, the level of human understanding of prostate cancer is limited, conventional treatment methods can not fundamentally remove malignant tumors, so the key mechanism of prostate cancer invasion and metastasis is identified, and the corresponding blocking pathway is found. It is particularly important to establish a set of effective treatment methods. Previous studies have shown that the short peptide B04, which specifically binds to high metastatic prostate cancer cell line PC3M, can inhibit the invasion and metastasis of PC3M cells. Therefore, we believe that its receptor on PC3M cells may be related to the metastasis of prostate cancer. In order to obtain the receptor of specific short peptide B04, western blot, Coomassie brilliant blue staining and silver staining were used to preliminarily localize the expression of specific short peptide B04 receptor. Then the identity information of the receptor protein was identified by affinity chromatography and mass spectrometry, and the subcellular localization of the receptor protein was analyzed by immunofluorescence cytochemical staining. The results showed that the receptor protein of short peptide B04 was the membrane protein of PC3M cells, and that ATP5B was one of the receptor proteins of short peptide B04, and that ATP5B was expressed in mitochondria and cell membrane of high metastatic prostate cancer cell line PC3M. It was found that the expression of ATP5B in the cell membrane of the metastasis associated protein of prostate cancer may be the key to its metastasis-related function.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25
【共引文献】
相关期刊论文 前1条
1 邸大琳;陈蕾;王丽娜;王洪伟;魏兵;王会东;鞠吉雨;;BCSC-1基因异位表达对MDA-MB-231侵袭能力影响及其机制探讨[J];中华肿瘤防治杂志;2014年06期
本文编号：2173614
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