甲状腺素受体β1基因抑制乳腺癌细胞增殖机制的研究

发布时间：2018-06-04 22:52

本文选题：乳腺癌 + TR1　；参考：《兰州大学》2017年硕士论文

【摘要】：目的:甲状腺素受体β1与人多种肿瘤的发生发展有关,但其在乳腺癌的作用及作用机理还不是很清楚。我们本次研究利用脂质体转染法使甲状腺素受体β1高表达于乳腺癌细胞株,检测该受体对乳腺癌细胞增殖能力的影响,并探索发挥该作用的机制。方法:挑选出两株乳腺癌细胞MCF-7和MDA-MB-231,用脂质体将THRB基因转染于乳腺癌细胞中,使乳腺癌细胞高表达甲状腺素受体β1,并利用免疫印记(western blotting/WB)的方法测定转染效率,确保转染有效。并在蛋白水平上检测PI3K-AKT-mTOR通路蛋白在三组中的改变。结果:免疫印记实验证实转染有效,MTT检测结果显示高表达TRβ1的细胞组增殖能力较空白对照组和转染空质粒组降低,Western Blotting检测蛋白水平:转染TRβ1组较其余两组p-AKT、p-mTOR表达低,总AKT、mTOR在三组之间无差异。加入配体T3后,随T3浓度的增高,p-AKT、p-mTOR的表达降低,表现出T3依赖性,总AKT、mTOR无差异。结论:TRβ1抑制乳腺癌细胞增殖,抑制PI3K/AKT/mTOR下游通路的AKT/mTOR的活化。在其配体T3存在的情况下,TRβ1对乳腺癌细胞的抑制作用增强,对p-AKT/p-mTOR通路抑制也增强。
[Abstract]:Objective: thyroxine receptor 尾 1 is associated with the occurrence and development of many kinds of human tumors, but its role and mechanism in breast cancer is not clear. In this study, thyroxine receptor 尾 1 was overexpressed in breast cancer cell line by liposome transfection, and the effect of thyroxine receptor on proliferation of breast cancer cell line was detected, and the mechanism of this effect was explored. Methods: two breast cancer cell lines, MCF-7 and MDA-MB-231, were selected. The THRB gene was transfected into breast cancer cells by liposome. The expression of thyroxine receptor 尾 1 was detected by Western blotting. The transfection efficiency was determined by Western blotting. The changes of PI3K-AKT-mTOR pathway proteins in the three groups were detected at the protein level. Results: the results of immunoimprinted assay showed that the proliferative ability of the cells with high expression of tr 尾 1 was lower than that of the blank control group and the empty plasmid group. The expression of p-AKTN p-mTOR in the transfected tr 尾 1 group was lower than that in the other two groups. There was no difference in total AKT mTOR among the three groups. When the ligand T3 was added, the expression of p-AKTO p-mTOR decreased with the increase of T3 concentration, and the expression of p-mTOR was T3 dependent. There was no difference in total AKTT mTOR. Conclusion: tr 尾 1 inhibits the proliferation of breast cancer cells and inhibits the activation of AKT/mTOR in the downstream pathway of PI3K/AKT/mTOR. The inhibitory effect of tr 尾 1 on breast cancer cells was enhanced in the presence of its ligand T3, and the inhibition of p-AKT/p-mTOR pathway was also enhanced.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9

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