LB-100增加低氧环境下肝细胞肝癌对索拉菲尼敏感性研究

发布时间：2018-05-09 01:12

本文选题：p-Smad3 + 凋亡　；参考：《浙江大学》2016年博士论文

【摘要】：背景和目的：肝细胞肝癌(HCC)是一种常见的肿瘤类型,预后较差。多激酶抑制剂索拉菲尼是临床上唯一获批用于肝癌系统性治疗的药物。然而,仅有少数病人对索拉菲尼敏感。低氧的微环境被认为是索拉菲尼耐药的原因之一。既往研究发现,LB-100,一种丝氨酸/苏氨酸蛋白酶磷酸酶2A (PP2A)抑制剂作用于肝细胞肝癌具有化疗增敏作用。本研究中,我们检测了LB-100是否具有增加肝癌细胞对索拉菲尼敏感性的作用。方法：使用Cell Counting Kit-8检测细胞活力；使用Annexin V-FITC Detection Kit试剂盒检测细胞凋亡；使用免疫印迹法检测细胞蛋白表达水平相对变化；使用Ser/Thr phosphatase assay kit I检测PP2A活性。在裸鼠模型上,比较索拉菲尼单用或与LB-100联合使用对肝细胞肝癌皮下移植瘤生长的影响。结果：仅在低氧环境中,LB-100能够增加索拉菲尼对肝癌细胞的抑制效果。LB-100能够显著地上调肝癌细胞中p-Smad3蛋白水平,p-Smad3水平的上调介导了LB-100的化疗增敏效应。LB-100下调了抗凋亡蛋白Bcl-2表达水平,增加了索拉菲尼诱导的肝癌凋亡。进一步地,我们证明LB-100诱导的p-Smad3过表达是通过抑制PP2A活性实现的。此外,在低氧环境下,LB-100诱导了p38 MAPK通路激活,这也增加了pSmad3依赖的Bcl-2下调及后续的凋亡。结论：低氧环境下,LB-100增加肝癌细胞对索拉菲尼的敏感性。这种化疗增敏作用是通过抑制PP2A,引起p-Smad3水平的升高实现的。升高的p-Smad3下调了Bcl-2,导致了肝癌细胞的凋亡增加。
[Abstract]:Background and purpose: hepatocellular carcinoma (HCC) is a common tumor type and has a poor prognosis. Sola Feeney, a multi kinase inhibitor, is the only drug for systematic treatment of liver cancer. However, only a few patients are sensitive to sorafeni. Hypoxic microenvironment is considered to be one of the reasons for Sola Feeney resistance. Now, LB-100, a serine / threonine protease phosphatase 2A (PP2A) inhibitor acts on chemotherapeutic sensitization in hepatocellular carcinoma. In this study, we detected whether LB-100 had an increase in the effect of hepatoma cells on Sola Feeney Min sensibility. Methods: Cell Counting Kit-8 was used to detect cell viability; Annexin V-FITC Dete was used. Ction Kit kits were used to detect cell apoptosis; the relative changes in cell protein expression were detected by Western blot; PP2A activity was detected using Ser/Thr phosphatase assay kit I. In nude mice, the effects of Sola Feeney alone or combined with LB-100 on the growth of hepatocellular carcinoma skin graft growth were compared. In the environment, LB-100 can increase the inhibitory effect of Sola Feeney on hepatoma cells.LB-100 can significantly increase the level of p-Smad3 protein in liver cancer cells. The up regulation of p-Smad3 level mediates the chemosensitivity effect of LB-100, which reduces the expression of anti apoptotic protein Bcl-2 and increases the apoptosis of liver cancer induced by Sao Rafini. Further, we LB-100 induced p-Smad3 overexpression was realized by inhibiting PP2A activity. In addition, in hypoxia environment, LB-100 induced the activation of p38 MAPK pathway, which also increased the Bcl-2 down regulation of pSmad3 dependent and subsequent apoptosis. Conclusion: in low oxygen environment, LB-100 increases the sensitivity of hepatoma cells to Sola Feeney. This chemosensitization effect is By inhibiting PP2A, the level of p-Smad3 was increased. The increase of p-Smad3 reduced Bcl-2, resulting in an increase in apoptosis of hepatoma cells.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7

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