铬促进前列腺癌细胞增殖效应的初步探索

发布时间：2019-02-28 14:12

【摘要】：目的通过筛选对促进前列腺癌PC3细胞系增值效应铬的价态、浓度及暴露时间。进一步探讨低剂量铬对该细胞系生物学行为的影响,初步探索其作用机制。方法通过MTT实验、CCK-8实验检测不同价态、不同浓度铬暴露不同时间后对前列腺癌PC3细胞系的影响,根据实验结果确定所需铬试剂,铬的剂量及暴露时间;根据MTT、CCK-8实验所得相关价态、浓度及作用时间,采用单克隆实验进一步验证低剂量铬暴露对前列腺癌PC3细胞的增殖效应;通过细胞迁移、细胞侵袭、细胞凋亡、细胞周期实验明确低剂量铬对前列腺癌PC3细胞系的生物学行为影响;小鼠成瘤实验进一步观察研究体内铬暴露对前列腺癌的影响;通过人类转录基因组测序进一步探索低剂量铬暴露促进前列腺癌细胞增殖效应的机制,通过Western Blot实验初步探索相关机制。结果MTT、CCK-8实验显示0.4umol/L铬酸钾暴露48小时后对前列腺癌细胞有显著的促进增殖效应。单克隆形成实验证实了这一结果。0.4umol/L铬酸钾暴露48小时后促进前列腺癌细胞迁徙、细胞侵袭,对细胞凋亡及细胞周期无明显作用。通过小鼠成瘤实验,5mg/kg/day的铬酸钾促进小鼠肿瘤增长。通过对经0.4μmol/L铬酸钾暴露48小时后的前列腺癌细胞与未经处理的前列腺癌细胞基因分析。结果显示经0.4umol/L铬酸钾处理后的标本与空白组标本存在差异表达基因,差异表现在生物学过程、细胞成分及分子机制,共表现出759个差异基因(387个上调基因,372个下调基因)。Western Blot结果显示经0.4umol/L铬酸钾处理后可促进PC3细胞Vimentin高表达,提示低剂量铬暴露促进前列腺癌PC3细胞EMT,进而证实低剂量铬暴露可促进前列腺癌细胞增殖、迁移及侵袭。结论低剂量铬暴露促进前列腺癌细胞增殖、迁移、侵袭,促进体内肿瘤生长。在生物学过程、细胞成分及分子机制等方面存在差异基因。Vimentin高表达,促进前列腺癌PC3细胞EMT,进而影响前列腺癌细胞增殖、迁移及侵袭。
[Abstract]:Objective to screen the valence, concentration and exposure time of chromium (Cr) to promote the proliferation of prostate cancer cell line PC3. To explore the effect of low dose chromium on the biological behavior of the cell line, and to explore its mechanism. Methods the effects of different valence states and different concentrations of chromium on prostate cancer cell line PC3 were detected by MTT test and CCK-8 assay. The dosage and exposure time of chromium reagent and chromium were determined according to the results of the experiment. The effect of low dose chromium exposure on the proliferation of prostate cancer PC3 cells was further verified by monoclonal assay according to the relative valence, concentration and action time of MTT,CCK-8 test. The biological behavior of prostate cancer cell line PC3 was determined by cell migration, invasion, apoptosis and cell cycle assay, and the effect of chromium exposure on prostate cancer was further observed in mice. The mechanism of promoting proliferation of prostate cancer cells by low-dose chromium exposure was further investigated by sequencing of human transcriptional genomes, and the related mechanisms were preliminarily explored by Western Blot assay. Results MTT,CCK-8 assay showed that 0.4umol/L potassium chromate significantly promoted the proliferation of prostate cancer cells 48 hours after exposure. After 48 hours of exposure to 0.4 umol / L potassium chromate, the migration and invasion of prostate cancer cells were promoted, but there was no obvious effect on cell apoptosis and cell cycle. 5mg/kg/day potassium chromate promoted tumor growth in mice by tumorigenicity test. The genes of prostate cancer cells and untreated prostate cancer cells exposed to 0.4 渭 mol / L potassium chromate for 48 hours were analyzed. The results showed that there were 759 differentially expressed genes in the biological process, cell composition and molecular mechanism between the samples treated with 0.4umol/L potassium chromate and those in the blank group, and a total of 759 up-regulated genes were found in the samples treated with potassium chromate. The results of 372 down-regulated genes). Western Blot showed that 0.4umol/L potassium chromate treatment could promote the high expression of Vimentin in PC3 cells, suggesting that low dose chromium exposure could promote EMT, expression in PC3 cells of prostate cancer. Furthermore, low-dose chromium exposure can promote proliferation, migration and invasion of prostate cancer cells. Conclusion low dose chromium exposure can promote the proliferation, migration and invasion of prostate cancer cells and promote the growth of prostate cancer cells in vivo. The high expression of vimentin promotes the proliferation, migration and invasion of prostate cancer cells by promoting EMT, in prostate cancer PC3 cells.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.25

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