喜树碱类似物FL118对人乳腺癌侵袭转移的影响及机制研究

发布时间：2018-03-30 15:08

本文选题：FL118　切入点：乳腺癌　出处：《青岛大学》2016年硕士论文

【摘要】：目的探讨喜树碱类似物FL118对人乳腺癌MDA-MB-231细胞侵袭转移能力的影响及其作用机制。方法以高转移的人乳腺癌MDA-MB-231细胞为研究对象,通过细胞增殖实验(MTT)观察FL118对肿瘤细胞活性及增殖情况的影响;流式细胞仪测定肿瘤细胞周期分布;细胞平板克隆形成实验、划痕修复实验及Transwell侵袭实验检测FL118干预后肿瘤细胞的平板克隆形成、转移及侵袭能力变化;实时荧光定量PCR(q PCR)、免疫细胞化学(ICC)和免疫印迹杂交(Western Blot)实验检测FL118对乳腺癌细胞survivin和上皮间质转化(EMT)相关信号分子E-钙黏蛋白(E-Cadherin)、波形蛋白(Vimentin)以及Wnt信号通路核内关键蛋白β-连环蛋白(β-catenin)及细胞周期素1(Cyclin Dl)的m RNA和蛋白水平的表达。结果FL118对MDA-MB-231细胞有增殖抑制作用,呈现一定的量效及时效关系(P0.05),且效果优于阳性对照组(P0.05);FL118使MDA-MB-231细胞有丝分裂受阻于S期(P0.05);FL118可降低MDA-MB-231细胞的平板克隆形成能力(P0.05);划痕实验及侵袭实验的结果表明FL118可以使MDA-MB-231细胞的转移和侵袭能力下降(P0.05);q PCR、ICC和Western Blot结果显示,FL118干预后,乳腺癌MDA-MB-231细胞E-Cadherin的m RNA及蛋白表达水平上升(P0.05),Vimentin、Survivin、β-catenin及Cyclin Dl的m RNA及蛋白表达水平下降(P0.05)。结论FL118能抑制乳腺癌MDA-MB-231细胞的增殖、克隆形成能力、转移和侵袭能力,并能提高E-Cadherin表达,降低Vimentin和Survivin表达,降低核内β-catenin及Cyclin Dl的表达,初步证明FL118抑制EMT的机制与Wnt通路有关。
[Abstract]:Objective to investigate the effect of camptothecin analogue FL118 on invasion and metastasis of human breast cancer MDA-MB-231 cells and its mechanism. Methods Human breast cancer MDA-MB-231 cells with high metastasis were studied. The effects of FL118 on the activity and proliferation of tumor cells were observed by cell proliferation assay. The cell cycle distribution was measured by flow cytometry. Scratch repair test and Transwell invasion assay were used to detect the formation, metastasis and invasion ability of tumor cells after FL118 intervention. Real-time fluorescence quantification of PCR(q, immunocytochemistry and Western blotanalysis to detect FL118 signaling molecules related to survivin and epithelial interstitial transformation of breast cancer cells, E-cadherin (vimentin) and Wnt signal pathway nucleus. Expression of m RNA and protein levels of 尾 -catenin and cyclin 1(Cyclin Dl. Results FL118 could inhibit the proliferation of MDA-MB-231 cells. The effect was better than that of the positive control group (P0.05 / FL-118). The mitosis of MDA-MB-231 cells was blocked in S phase (P0.05 / FL-118), and the ability of MDA-MB-231 cells to form flat cells was decreased by P0.05FL-118. The results of scratch test and invasion experiment showed that FL118 could inhibit the formation of MDA-MB-231 cells. In order to decrease the metastasis and invasion ability of MDA-MB-231 cells, the results of Western Blot and P0.05 Blot showed that after the intervention of FL-118, The expression level of m RNA and protein in E-Cadherin of breast cancer MDA-MB-231 cells increased. The expression levels of m RNA and protein of Vimentinus survivin, 尾 -catenin and Cyclin D1 decreased. Conclusion FL118 can inhibit the proliferation, clone formation, metastasis and invasion of breast cancer MDA-MB-231 cells, and increase the expression of E-Cadherin. The expression of Vimentin and Survivin and the expression of 尾 -catenin and Cyclin D1 in the nucleus were decreased, which suggested that the mechanism of FL118 inhibiting EMT was related to the Wnt pathway.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R96

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