發(fā)布時間：2018-11-11 09:58

【摘要】：【目的】研究防己諾林堿對三陰性乳腺癌MDA-MB-231細胞的促進凋亡、抑制PI3K/AKT/mTOR通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表達,抑制荷瘤裸鼠腫瘤的生長,探討防己諾林堿對三陰性乳腺癌的抗腫瘤機制�！痉椒ā矿w外培養(yǎng)三陰性乳腺癌細胞MDA-MB-231,刃天青法檢測防己諾林堿對三陰性乳腺癌細胞MDA-MB-231的抑制作用,計算其IC50,確定防己諾林堿的有效濃度;6孔板細胞劃痕實驗檢測防己諾林堿抑制三陰性乳腺癌細胞MDA-MB-231的遷徙能力;細胞流式技術(shù)檢測防己諾林堿促進細胞的凋亡作用;Western Blot檢測PI3K/AKT/mTOR通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表達,從而確定防己諾林堿對三陰性乳腺癌細胞MDA-MB-231蛋白表達的影響;荷瘤裸鼠實驗探討防己諾林堿對三陰性乳腺癌腫瘤生長的抑制作用�！窘Y(jié)果】防己諾林堿可以抑制三陰性乳腺癌細胞MDA-MB-231的活力,并測得IC50為6.25μmol/L;6孔板細胞劃痕實驗結(jié)果表明防己諾林堿還可以抑制MDA-MB-231細胞的遷徙能力;隨著濃度升高,抑制作用結(jié)果明顯;細胞流式技術(shù)表明防己諾林堿可以促進MDA-MB-231細胞的凋亡,且隨著濃度增加,凋亡率呈正相關(guān);Western Blot實驗結(jié)果表明防己諾林堿可調(diào)控PI3K/AKT/mTOR信號通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表達,且表達結(jié)果受濃度影響,隨濃度升高,蛋白表達下降;裸鼠成瘤實驗結(jié)果表明防己諾林堿可抑制三陰性乳腺癌腫瘤的生長�！窘Y(jié)論】防己諾林堿可以降低MDA-MB-231細胞的活力,加速細胞的凋亡,抑制細胞遷徙,下調(diào)PI3K/AKT/mTOR信號通路中非磷酸化及磷酸化PI3K、AKT、mTOR表達,抑制TNBC細胞的生長、增殖、遷移,達到抑制腫瘤的生長效果。
[Abstract]:[objective] to study the effect of Tetrandrine on the apoptosis of three-negative breast cancer MDA-MB-231 cells, inhibit the expression of PI3K,AKT,mTOR and phosphorylated PI3K,AKT,mTOR in PI3K/AKT/mTOR pathway, and inhibit the growth of tumor-bearing nude mice. To investigate the anti-tumor mechanism of tri-negative breast cancer treated with fanginolinine. [methods] the inhibitory effect of fanginolinine on MDA-MB-231 of tri-negative breast cancer cells was detected by MDA-MB-231, assay in vitro. The effective concentration of Tetrandrine was determined by calculating its IC50,. 6-well plate cell scratch assay was used to detect the inhibition of MDA-MB-231 migration in triple-negative breast cancer cells, and cell flow cytometry was used to detect the effect of fenpentine on the apoptosis of breast cancer cells. Western Blot was used to detect the expression of PI3K,AKT,mTOR and phosphorylated PI3K,AKT,mTOR protein in PI3K/AKT/mTOR pathway, so as to determine the effect of Tetrandrine on the expression of MDA-MB-231 protein in triple-negative breast cancer cells. To investigate the inhibitory effect of Tetrandrine on the growth of tri-negative breast cancer cells in nude mice. [results] the activity of MDA-MB-231 in tri-negative breast cancer cells was inhibited and the IC50 was measured to be 6.25 渭 mol/L;. The results of scratch test showed that Tetrandrine could also inhibit the migration ability of MDA-MB-231 cells, and the inhibitory effect was obvious with the increase of concentration. Flow cytometry showed that Tetrandrine could promote the apoptosis of MDA-MB-231 cells, and the apoptosis rate was positively correlated with the increase of concentration. Western Blot results showed that Tetrandrine could regulate the expression of PI3K,AKT,mTOR and phosphorylated PI3K,AKT,mTOR protein in PI3K/AKT/mTOR signaling pathway, and the expression of PI3K,AKT,mTOR and phosphorylated PI3K,AKT,mTOR protein was affected by the concentration, and the protein expression decreased with the increase of the concentration. The results of tumorigenesis in nude mice showed that Tetrandrine could inhibit the growth of three-negative breast cancer tumor. [conclusion] Tetrandrine can reduce the activity of MDA-MB-231 cells, accelerate cell apoptosis and inhibit cell migration. The expression of non-phosphorylation and phosphorylated PI3K,AKT,mTOR in PI3K/AKT/mTOR signaling pathway was down-regulated, and the growth, proliferation and migration of TNBC cells were inhibited.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R737.9

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