防己诺林碱对三阴性乳腺癌抗肿瘤机制的研究

发布时间：2018-11-11 09:58

【摘要】：【目的】研究防己诺林碱对三阴性乳腺癌MDA-MB-231细胞的促进凋亡、抑制PI3K/AKT/mTOR通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表达,抑制荷瘤裸鼠肿瘤的生长,探讨防己诺林碱对三阴性乳腺癌的抗肿瘤机制。【方法】体外培养三阴性乳腺癌细胞MDA-MB-231,刃天青法检测防己诺林碱对三阴性乳腺癌细胞MDA-MB-231的抑制作用,计算其IC50,确定防己诺林碱的有效浓度;6孔板细胞划痕实验检测防己诺林碱抑制三阴性乳腺癌细胞MDA-MB-231的迁徙能力;细胞流式技术检测防己诺林碱促进细胞的凋亡作用;Western Blot检测PI3K/AKT/mTOR通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表达,从而确定防己诺林碱对三阴性乳腺癌细胞MDA-MB-231蛋白表达的影响;荷瘤裸鼠实验探讨防己诺林碱对三阴性乳腺癌肿瘤生长的抑制作用。【结果】防己诺林碱可以抑制三阴性乳腺癌细胞MDA-MB-231的活力,并测得IC50为6.25μmol/L;6孔板细胞划痕实验结果表明防己诺林碱还可以抑制MDA-MB-231细胞的迁徙能力;随着浓度升高,抑制作用结果明显;细胞流式技术表明防己诺林碱可以促进MDA-MB-231细胞的凋亡,且随着浓度增加,凋亡率呈正相关;Western Blot实验结果表明防己诺林碱可调控PI3K/AKT/mTOR信号通路中PI3K、AKT、mTOR及磷酸化PI3K、AKT、mTOR蛋白的表达,且表达结果受浓度影响,随浓度升高,蛋白表达下降;裸鼠成瘤实验结果表明防己诺林碱可抑制三阴性乳腺癌肿瘤的生长。【结论】防己诺林碱可以降低MDA-MB-231细胞的活力,加速细胞的凋亡,抑制细胞迁徙,下调PI3K/AKT/mTOR信号通路中非磷酸化及磷酸化PI3K、AKT、mTOR表达,抑制TNBC细胞的生长、增殖、迁移,达到抑制肿瘤的生长效果。
[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.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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