前列腺癌移植小鼠模型的建立和前列腺癌高侵袭性和低侵袭性细胞的体内外筛选

发布时间：2018-10-20 20:35

【摘要】： 目的建立前列腺癌移植性转移模型,筛选同一标本来源的高侵袭性和低侵袭性前列腺癌细胞,为研究前列腺癌的转移分子标志奠定基础。方法1、采用皮下、前列腺原位、肾包膜下法对BNX或NOD/SCIDX小鼠接种前列腺癌细胞悬液,观察各种方法的成瘤及转移情况;2、RT-PCR方法分析转移相关基因MAPK4、SELM、IL-6、Stanniocalcin 2、Serpine1、Caveline-1、Laminin4、AI793338在前列腺癌原发灶和转移灶组织中的表达情况;3、采用Transwell对前列腺癌自发转移模型成瘤的细胞进行体外侵袭实验,分离高侵袭性和低侵袭性细胞,用RT-PCR方法分析转移相关基因MMP-2、TIMP-2表达。结果1、成功建立了前列腺癌细胞系原位移植自发转移模型,分离获得了前列腺癌肺、淋巴结转移细胞;2、RT-PCR结果证实MAPK4、SELM、IL-6在前列腺癌转移组织中表达量较原发肿瘤表达量低,Stanniocalcin 2、Serpine1、Laminin4在前列腺癌转移组织中表达量较原发肿瘤表达量高;3、采用Transwell实验筛选出高侵袭性和低侵袭性细胞株,借助细胞生长曲线观察细胞的体外增殖情况和非贴壁依赖性生长能力,结果表明:高侵袭亚系比低侵袭亚系具有明显生长优势,且非贴壁依赖性生长能力也较强。RT-PCR结果证实MMP-2在高侵袭性细胞株中表达量增强,TIMP-2在高侵袭性细胞株中表达量低。结论通过细胞悬液原位注射法成功建立了前列腺癌自发转移模型。Transwell筛选出前列腺癌高侵袭性和低侵袭性细胞株。本研究为前列腺癌转移机制奠定了基础。
[Abstract]:Objective to establish a transplant metastasis model of prostate cancer and screen highly invasive and low invasive prostate cancer cells from the same specimen, so as to lay a foundation for studying the molecular markers of prostate cancer metastasis. Methods 1. BNX or NOD/SCIDX mice were inoculated with prostate cancer cell suspension by subcutaneous, in situ and subcapsular method. The tumorigenesis and metastasis of various methods were observed, the expression of metastasis related gene MAPK4,SELM,IL-6,Stanniocalcin _ 2, Serpine _ 1, Caveline-1, Laminine _ 4 and AI793338 in primary and metastatic tissues of prostate cancer was analyzed by RT-PCR. 3, Transwell was used to investigate the invasion of tumor cells from spontaneous metastasis model of prostate cancer in vitro. High invasive and low invasive cells were isolated, and MMP-2,TIMP-2 expression of metastasis related genes was analyzed by RT-PCR method. Results 1. The spontaneous metastasis model of prostate cancer cell line was successfully established and prostate cancer lung was isolated. The results of RT-PCR confirmed that the expression of MAPK4,SELM,IL-6 was lower in the metastatic tissue of prostate cancer than that in the primary tumor, and the expression of Stanniocalcin _ 2Serpine _ 1 laminin4 was higher in the metastatic tissue of prostate cancer than that in the primary tumor. 3, the high invasion was screened by Transwell assay. Invasive and low invasive cell lines, Cell growth curve was used to observe cell proliferation and adhesion-independent growth ability in vitro. The results showed that high invasive sublines had obvious growth advantages over low invasive sublines. The results of RT-PCR showed that the expression of MMP-2 was increased in highly invasive cell lines, and the expression of TIMP-2 was low in highly invasive cell lines. Conclusion the spontaneous metastasis model of prostate cancer was successfully established by in situ injection of cell suspension and Transwell was used to screen high invasive and low invasive prostate cancer cell lines. This study laid a foundation for the metastasis mechanism of prostate cancer.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R737.25;R-332

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