本文選題：FL118　切入點(diǎn)：乳腺癌　出處：《青島大學(xué)》2016年碩士論文

【摘要】：目的探討喜樹堿類似物FL118對(duì)人乳腺癌MDA-MB-231細(xì)胞侵襲轉(zhuǎn)移能力的影響及其作用機(jī)制。方法以高轉(zhuǎn)移的人乳腺癌MDA-MB-231細(xì)胞為研究對(duì)象,通過(guò)細(xì)胞增殖實(shí)驗(yàn)(MTT)觀察FL118對(duì)腫瘤細(xì)胞活性及增殖情況的影響;流式細(xì)胞儀測(cè)定腫瘤細(xì)胞周期分布;細(xì)胞平板克隆形成實(shí)驗(yàn)、劃痕修復(fù)實(shí)驗(yàn)及Transwell侵襲實(shí)驗(yàn)檢測(cè)FL118干預(yù)后腫瘤細(xì)胞的平板克隆形成、轉(zhuǎn)移及侵襲能力變化;實(shí)時(shí)熒光定量PCR(q PCR)、免疫細(xì)胞化學(xué)(ICC)和免疫印跡雜交(Western Blot)實(shí)驗(yàn)檢測(cè)FL118對(duì)乳腺癌細(xì)胞survivin和上皮間質(zhì)轉(zhuǎn)化(EMT)相關(guān)信號(hào)分子E-鈣黏蛋白(E-Cadherin)、波形蛋白(Vimentin)以及Wnt信號(hào)通路核內(nèi)關(guān)鍵蛋白β-連環(huán)蛋白(β-catenin)及細(xì)胞周期素1(Cyclin Dl)的m RNA和蛋白水平的表達(dá)。結(jié)果FL118對(duì)MDA-MB-231細(xì)胞有增殖抑制作用,呈現(xiàn)一定的量效及時(shí)效關(guān)系(P0.05),且效果優(yōu)于陽(yáng)性對(duì)照組(P0.05);FL118使MDA-MB-231細(xì)胞有絲分裂受阻于S期(P0.05);FL118可降低MDA-MB-231細(xì)胞的平板克隆形成能力(P0.05);劃痕實(shí)驗(yàn)及侵襲實(shí)驗(yàn)的結(jié)果表明FL118可以使MDA-MB-231細(xì)胞的轉(zhuǎn)移和侵襲能力下降(P0.05);q PCR、ICC和Western Blot結(jié)果顯示,FL118干預(yù)后,乳腺癌MDA-MB-231細(xì)胞E-Cadherin的m RNA及蛋白表達(dá)水平上升(P0.05),Vimentin、Survivin、β-catenin及Cyclin Dl的m RNA及蛋白表達(dá)水平下降(P0.05)。結(jié)論FL118能抑制乳腺癌MDA-MB-231細(xì)胞的增殖、克隆形成能力、轉(zhuǎn)移和侵襲能力,并能提高E-Cadherin表達(dá),降低Vimentin和Survivin表達(dá),降低核內(nèi)β-catenin及Cyclin Dl的表達(dá),初步證明FL118抑制EMT的機(jī)制與Wnt通路有關(guān)。
[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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R96

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