胰岛素对前列腺癌细胞的作用及机制

发布时间：2018-03-13 06:07

本文选题：前列腺肿瘤　切入点：胰岛素　出处：《山东大学》2014年硕士论文　论文类型：学位论文

【摘要】：目的：建立雄激素非依赖前列腺癌细胞系,观察胰岛素对三种前列腺癌细胞系LNCaP-AI、PC-3)增殖、活力、迁移与侵袭能力的影响,并探讨其可能机制。方法：本研究分为两部分。(1)去雄激素环境下连续培养前列腺癌LNCaP细胞,建立雄激素非依赖前列腺癌细胞系LNCaP-AI;(2)胰岛素分别刺激LNCaP、 LNCaP-AI、PC-3三种细胞,采用实时荧光定量聚合酶链反应(Real-Time PCR)、蛋白免疫印迹(Western Blotting)、免疫细胞化学(immunocytochemistry, ICC)法检测胰岛素受体表达变化；细胞计数法、四唑盐法(MTT)、细胞划痕试验、细胞侵袭试验验(Transwell)分别检测细胞接受刺激前后增殖、活力、迁移和侵袭能力的变化；Western Blotting检测细胞接受刺激前后Ras、ERK1/2、MEK1/2蛋白的表达变化以观察MAPK信号通路的活性变化。结果： (1)6个月后成功建立LNCaP-AI细胞系。该细胞可在去雄环境下正常成长、传代、冻存及复苏,与LNCaP相比,细胞形态发生改变,雄激素受体表达升高,胰岛素受体表达无明显变化；(2)胰岛素刺激可使三种细胞的胰岛素受体在mRNA及蛋白水平表达均增加；胰岛素对LNCaP细胞的增殖、活力、迁移及侵袭均有所抑制,对PC-3细胞有明显促进作用,而LNCaP-AI细胞在胰岛素刺激前后增殖、活力、迁移及侵袭均无明显变化；胰岛素可降低LNCaP细胞Ras的表达及ERK、MEK的磷酸化水平,促进PC-3细胞Ras的表达及ERK、MEK的磷酸化水平,而对LNCaP-AI细胞无明显作用。结论：胰岛素可通过胰岛素受体影响Ras-MEK-ERK信号通路的活化从而抑制LNCaP细胞的增殖、活力、迁移及侵袭,促进PC-3细胞的增殖、活力、迁移及侵袭,但对LNCaP-AI细胞无明显作用。
[Abstract]:Objective:. The androgen independent prostate cancer cell line was established to observe the effects of insulin on the proliferation, activity, migration and invasion of three prostate cancer cell lines LNCaP-AIPC-3, and to explore its possible mechanism. Methods:. The present study was divided into two parts: 1) LNCaP cells of prostate cancer were cultured continuously in androgen free environment, androgen independent prostate cancer cell line LNCaP-AIP-2) was induced by insulin, and three kinds of cells, LNCaP, LNCaP-AIPC-3, were induced by insulin, respectively. The expression of insulin receptor was detected by real-time fluorescence quantitative polymerase chain reaction Real-Time PCR, Western blotting, immunocytochemistryassay (ICCC), cell counting, tetrazolium salt method and cell scratch test. Cell invasion assay was used to detect the changes of cell proliferation, activity, migration and invasion ability before and after stimulation. Western Blotting was used to detect the expression of Raser ERK1 / 2 MEK1 / 2 protein before and after stimulation to observe the activity of MAPK signaling pathway. Results:. After 6 months, LNCaP-AI cell line was successfully established. The cell line could grow, subculture, cryopreservation and resuscitation in deandrogenic environment. Compared with LNCaP, the cell morphology was changed, androgen receptor expression was increased. Insulin receptor expression did not change significantly. Insulin stimulation increased the expression of insulin receptor at the level of mRNA and protein, and insulin inhibited the proliferation, activity, migration and invasion of LNCaP cells. LNCaP-AI cells showed no significant changes in proliferation, activity, migration and invasion before and after insulin stimulation, and insulin could decrease the expression of Ras and phosphorylation of ERK in LNCaP cells. Promote the expression of Ras and phosphorylation of ERK in PC-3 cells, but have no effect on LNCaP-AI cells. Conclusion:. Insulin can inhibit the proliferation, activity, migration and invasion of LNCaP cells by affecting the activation of Ras-MEK-ERK signaling pathway, and promote the proliferation, activity, migration and invasion of PC-3 cells, but it has no obvious effect on LNCaP-AI cells.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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