重组蛋白rLj-112对人非小细胞肺癌A549细胞增殖、迁移和凋亡的影响及其机制研究  

The Effects and Mechanism Studies of Recombinant Protein rLj-112 on Proliferation, Migration and Apoptosis of Human Non-small Cell Lung Cancer A549 Cells

作　　者：郑媛媛[1] 王继红[1] 李庆伟[1] 

机构地区：[1]辽宁师范大学生命科学学院,大连116029

出　　处：《中国细胞生物学学报》2017年第12期1528-1535,共8页Chinese Journal of Cell Biology

基　　金：国家高技术研究发展规划(863计划)(批准号:SS2014AA091602);国家自然科学基金(批准号:30770297)资助的课题~~

摘　　要：该文研究了重组蛋白r Lj-112对人非小细胞肺癌A549细胞增殖、迁移和凋亡的影响及其作用机制。采用MTT(四甲基偶氮唑蓝)法检测r Lj-112对人非小细胞肺癌A549细胞增殖的影响,采用Transwell方法检测r Lj-112对A549细胞迁移和侵袭的影响,Hoechst和吉姆萨染色的方法检测r Lj-112对A549细胞凋亡的影响;采用Western blot法检测r Lj-112对A549细胞信号通路相关蛋白质水平的影响。结果表明,r Lj-112能够抑制A549细胞的增殖,半抑制浓度IC50值为1.64μmol/L;r Lj-112能抑制A549细胞的迁移和侵袭并呈剂量依赖性;r Lj-112能诱导A549细胞的凋亡;r Lj-112处理过的细胞cleaved-caspase3蛋白质和cleaved-PARP蛋白质水平升高,证明r Lj-112通过caspase3/PARP途径执行对A549细胞的凋亡的诱导,p-AKT、p-PI3K和p-Erk1/2的水平下降,提示r Lj-112的作用方式与PI3K/AKT相关信号通路有关。We studied the ability of r Lj-112 protein from the Lamprey Japanica on anti-tumor of A549 cells and its regulatory mechanism in this study. We used MTT assay to detect cell proliferation. Giemsa staining assay and Hoechst 33258 staining were employed to examine cell apoptosis of A549 cells after Lj-112 treated. Western blot was used to examine cell apoptosis related signal proteins. It was found that after treated with r Lj-112, the proliferation of A549 cells was inhibited and IC50 value was 1.64 μmol/L. The inhibition effect is in a dosedependent manner. The migration assay showed that after treated with different concentrations of r Lj-112 proteins the inhibition effect was in a dose-dependent manner. The same situation occurred in the invasion test. The Hoechst 33258 staining experiment showed r Lj-112 could induce the apoptosis of A549 cell. The Western blot analysisshowed that cleaved-caspase3 protein and cleaved-PARP protein were improved in A549 cells treated with rLj-112 and demonstrated that rLj-112 induced apoptosis of A549 cells through caspase3/PARP pathway. The levels of p-AKT, p-PI3 K and p-Erk1/2 were decreased in the rLj-112 treated A549 cells, demonstrated that rLj-112 reduced AKT/PI3 K pathway in A549 cells.

关 键 词：非小细胞肺癌 增殖 细胞迁移和侵袭 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]