Connexin 43对非小细胞肺癌细胞吉非替尼原发性耐药的影响

发布时间：2018-05-08 09:58

  本文选题：缝隙连接蛋白 + 非小细胞肺癌　； 参考：《重庆医科大学学报》2017年11期

【摘要】：目的:探讨缝隙连接蛋白43(connexin 43,Cx43)对非小细胞肺癌(non-small-cell lung cancer,NSCLC)吉非替尼原发性耐药的影响。方法:MTT法测定吉非替尼对3株非突变型EGFR(WT-EGFR)NSCLC细胞系A549、H1299、Calu3细胞的半数抑制浓度(IC50);Western blot观察Cx43、磷酸化Akt(phospho-Akt,p-Akt)蛋白在上述细胞系中的表达水平;"Parachute"法检测细胞缝隙连接功能(gap junction intercellular communication,GJIC);免疫荧光法检测Cx43蛋白在细胞中的定位。结果:吉非替尼作用于Calu3、A549、H1299细胞的IC50分别为(0.064±0.011)μmol/L、(13.64±0.015)μmol/L、(20.054±0.012)μmol/L,即A549、H1299细胞对吉非替尼原发性耐药;A549、H1299细胞中Cx43、p-Akt蛋白水平显著高于Calu3细胞(P0.01);A549、H1299细胞有荧光传递现象,且Cx43蛋白定位在A549、H1299细胞膜上,而Calu3细胞无荧光传递现象,且Cx43主要表达于细胞质上。结论:NSCLC细胞膜上调的Cx43及其组成的GJIC可促进细胞对吉非替尼的原发性耐药,其机制可能与其激活PI3K/Akt信号通路有关。
[Abstract]:Objective: to investigate the effect of gap junction protein (43(connexin 43) on primary drug resistance of non-small cell lung cancer (NSCLC) in patients with non small cell lung cancer (NSCC). Methods the expression of Cx43, phosphorylated Aktophospho-Aktna-p-Akt protein in three non-mutant EGFR(WT-EGFR)NSCLC cell lines A549H 129- Calu3 cell line was detected by Western blot, the expression of Cx43, phosphorylated Aktphospho-Aktophospho-Aktnp-Akt protein was detected by "Parachute" method, the gap junction intercellular communication was detected by "Parachute" method, and the expression of Cx43, phosphorylated Aktophospho-Aktophosphor-Aktp-Akt protein was detected by "Parachute" method. The localization of Cx43 protein in cells was detected by immunofluorescence assay. Results: the levels of IC50 in Cx43H1299 cells were significantly higher than those in Calu3 cells (P0.01, A549H1299 cells), and the fluorescence transfer of Cx43 protein was observed in A549H1299 cells. The results showed that the expression of Cx43 protein in A549H1299 cells was significantly higher than that in P0.01A549H1299 cells, and the Cx43 protein was localized on the A549H1299 cell membrane, and the expression of Cx43 protein was located on the A549H1299 cell membrane, and the protein level of A549H1299 cells was significantly higher than that of the Cx43p-Akt cells of Cx43p-Akt cells. The results showed that the expression of Cx43 protein was localized on the cell membrane of A549H1299 cells, and the expression of Cx43 protein was localized on the A549H1299 cell membrane of A549H1299 cells. However, there was no fluorescence transfer in Calu3 cells, and Cx43 was mainly expressed in cytoplasm. Conclusion the up-regulated Cx43 and its component GJIC can promote the primary resistance to gifitinib, and the mechanism may be related to the activation of PI3K/Akt signaling pathway.
【作者单位】： 广西医科大学药学院药理学教研室;
【基金】：国家自然科学基金资助项目(编号:81260324) 教育部高等学校博士学科点专项科研基金新教师类资助项目(编号:20124503120008)
【分类号】：R734.2
，

本文编号：1860946