【摘要】： 目的建立前列腺癌移植性轉(zhuǎn)移模型,篩選同一標(biāo)本來(lái)源的高侵襲性和低侵襲性前列腺癌細(xì)胞,為研究前列腺癌的轉(zhuǎn)移分子標(biāo)志奠定基礎(chǔ)。 方法1、采用皮下、前列腺原位、腎包膜下法對(duì)BNX或NOD/SCIDX小鼠接種前列腺癌細(xì)胞懸液,觀察各種方法的成瘤及轉(zhuǎn)移情況;2、RT-PCR方法分析轉(zhuǎn)移相關(guān)基因MAPK4、SELM、IL-6、Stanniocalcin 2、Serpine1、Caveline-1、Laminin4、AI793338在前列腺癌原發(fā)灶和轉(zhuǎn)移灶組織中的表達(dá)情況;3、采用Transwell對(duì)前列腺癌自發(fā)轉(zhuǎn)移模型成瘤的細(xì)胞進(jìn)行體外侵襲實(shí)驗(yàn),分離高侵襲性和低侵襲性細(xì)胞,用RT-PCR方法分析轉(zhuǎn)移相關(guān)基因MMP-2、TIMP-2表達(dá)。 結(jié)果1、成功建立了前列腺癌細(xì)胞系原位移植自發(fā)轉(zhuǎn)移模型,分離獲得了前列腺癌肺、淋巴結(jié)轉(zhuǎn)移細(xì)胞;2、RT-PCR結(jié)果證實(shí)MAPK4、SELM、IL-6在前列腺癌轉(zhuǎn)移組織中表達(dá)量較原發(fā)腫瘤表達(dá)量低,Stanniocalcin 2、Serpine1、Laminin4在前列腺癌轉(zhuǎn)移組織中表達(dá)量較原發(fā)腫瘤表達(dá)量高;3、采用Transwell實(shí)驗(yàn)篩選出高侵襲性和低侵襲性細(xì)胞株,借助細(xì)胞生長(zhǎng)曲線觀察細(xì)胞的體外增殖情況和非貼壁依賴性生長(zhǎng)能力,結(jié)果表明:高侵襲亞系比低侵襲亞系具有明顯生長(zhǎng)優(yōu)勢(shì),且非貼壁依賴性生長(zhǎng)能力也較強(qiáng)。RT-PCR結(jié)果證實(shí)MMP-2在高侵襲性細(xì)胞株中表達(dá)量增強(qiáng),TIMP-2在高侵襲性細(xì)胞株中表達(dá)量低。 結(jié)論通過(guò)細(xì)胞懸液原位注射法成功建立了前列腺癌自發(fā)轉(zhuǎn)移模型。Transwell篩選出前列腺癌高侵襲性和低侵襲性細(xì)胞株。本研究為前列腺癌轉(zhuǎn)移機(jī)制奠定了基礎(chǔ)。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R737.25;R-332

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本文編號(hào)：2284324