双氢青蒿素对PC-3和DU145细胞迁移和侵袭能力的影响及机制

发布时间：2018-09-11 10:18

【摘要】：目的：探讨双氢青蒿素（DHA）对PC-3和DU145细胞迁移和侵袭能力的影响及其可能机制。方法：第一部分：不同浓度（0μmol·L-1、25μmol·L-1、50μmol·L-1和100μmol·L-1）的DHA分别作用于PC-3和DU145细胞48h后，运用细胞划痕实验和侵袭实验检测细胞的迁移和侵袭能力。提取各组细胞的总mRNA和蛋白，分别进行荧光定量PCR和Western blot实验，检测Vimentin和CDH1的转录和表达水平。第二部分：采用人前列腺癌PC-3和DU145细胞建立裸鼠种植瘤模型，18只模型鼠随机分为对照组、DMSO组（二甲基亚砜1mL/kg）、DHA组(DHA200μmol/kg），经13天干预后，免疫组织（SP）化学法检测侵袭相关基因Vimentin和CDH1的表达水平。结果：不同浓度（25μmol·L-1、50μmol·L-1和100μmol·L-1）的DHA均能显著抑制PC-3和DU145迁移和侵袭能力（P0.05），与对照组比较，不同浓度的DHA均能显著抑制Vimentin和促进CDH1的转录和表达水平（P0.05）。25μmol·L-1的DHA抑制Vimentin和促进CDH1蛋白的表达最为明显。免疫组化结果显示，在两种细胞中，，DHA组CDH1的表达均高于对照组和溶剂组（P0.05）；Vimentin的表达均低于对照组和溶剂组（P0.05）。结论：不同浓度的DHA干预后能显著抑制PC-3和DU145细胞的迁移和侵袭能力，而在体肿块生长速度受抑制进一步证实DHA具有抑制癌细胞迁移和侵袭能力。而从免疫印迹和免疫组化结果我们推测DHA可能通过干扰Vimentin和CDH1蛋白的表达相关而起到抑制癌细胞迁移和侵袭的效果。
[Abstract]:Aim: to investigate the effect of dihydroartemisinin (DHA) on migration and invasion of PC-3 and DU145 cells and its possible mechanism. Methods: in the first part, the cells of PC-3 and DU145 were treated with different concentrations of DHA (0 渭 mol L -1, 25 渭 mol L -1, 50 渭 mol L -1 and 100 渭 mol L -1) for 48 h, respectively. Cell scratch assay and invasion assay were used to detect the migration and invasion ability of the cells. The total mRNA and protein were extracted from the cells of each group, and the transcription and expression levels of Vimentin and CDH1 were detected by fluorescence quantitative PCR and Western blot assay. Part two: PC-3 and DU145 cells were used to establish implanted tumor model in nude mice. Eighteen nude mice were randomly divided into two groups: control group (1mL/kg) (DHA200 渭 mol/kg). Immunohistochemical (SP) method was used to detect the expression of Vimentin and CDH1. Results: DHA at different concentrations (25 渭 mol L-1 and 100 渭 mol L-1) significantly inhibited the migration and invasion of PC-3 and DU145 (P0.05), compared with the control group. DHA at different concentrations could significantly inhibit Vimentin and promote the transcription and expression of CDH1 (P0.05) .25 渭 mol L-1 DHA could inhibit Vimentin and promote CDH1 protein expression. Immunohistochemical results showed that the expression of CDH1 in DHA group was higher than that in control group and solvent group (P0.05), and the expression of Vimentin was lower than that in control group and solvent group (P0.05). Conclusion: different concentrations of DHA can significantly inhibit the migration and invasion of PC-3 and DU145 cells, and the inhibition of tumor growth rate in vivo further confirms that DHA can inhibit the migration and invasion of cancer cells. From the results of immunoblotting and immunohistochemistry we speculate that DHA may inhibit the migration and invasion of cancer cells by interfering with the expression of Vimentin and CDH1 proteins.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.25

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