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雙氫青蒿素對PC-3和DU145細胞遷移和侵襲能力的影響及機制

發(fā)布時間:2018-09-11 10:18
【摘要】:目的:探討雙氫青蒿素(DHA)對PC-3和DU145細胞遷移和侵襲能力的影響及其可能機制。 方法:第一部分:不同濃度(0μmol·L-1、25μmol·L-1、50μmol·L-1和100μmol·L-1)的DHA分別作用于PC-3和DU145細胞48h后,運用細胞劃痕實驗和侵襲實驗檢測細胞的遷移和侵襲能力。提取各組細胞的總mRNA和蛋白,分別進行熒光定量PCR和Western blot實驗,檢測Vimentin和CDH1的轉錄和表達水平。第二部分:采用人前列腺癌PC-3和DU145細胞建立裸鼠種植瘤模型,18只模型鼠隨機分為對照組、DMSO組(二甲基亞砜1mL/kg)、DHA組(DHA200μmol/kg),經13天干預后,免疫組織(SP)化學法檢測侵襲相關基因Vimentin和CDH1的表達水平。 結果:不同濃度(25μmol·L-1、50μmol·L-1和100μmol·L-1)的DHA均能顯著抑制PC-3和DU145遷移和侵襲能力(P0.05),與對照組比較,不同濃度的DHA均能顯著抑制Vimentin和促進CDH1的轉錄和表達水平(P0.05)。25μmol·L-1的DHA抑制Vimentin和促進CDH1蛋白的表達最為明顯。免疫組化結果顯示,在兩種細胞中,,DHA組CDH1的表達均高于對照組和溶劑組(P0.05);Vimentin的表達均低于對照組和溶劑組(P0.05)。 結論:不同濃度的DHA干預后能顯著抑制PC-3和DU145細胞的遷移和侵襲能力,而在體腫塊生長速度受抑制進一步證實DHA具有抑制癌細胞遷移和侵襲能力。而從免疫印跡和免疫組化結果我們推測DHA可能通過干擾Vimentin和CDH1蛋白的表達相關而起到抑制癌細胞遷移和侵襲的效果。
[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.
【學位授予單位】:重慶醫(yī)科大學
【學位級別】:碩士
【學位授予年份】:2014
【分類號】:R737.25

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