Home > Browse Issues > Vol.35 No.11
The Effect of RhoA Protein on Mycobacterium tuberculosis Invasion of A549 Cells
Shao Shengwen1, Zhou Hongchang1, Xu Baiying1, Zhang Hui1, Xue Lijun2*
1Laboratory of Innovation, Medical School of Huzhou Teachers College, Huzhou 313000, China;2Department of Medical Oncology,Nanjing General Hospital of Nanjing Military Command, Nanjing 210002, China
Abstract: This paper was to investigate the role of RhoA protein in the process of Mycobacterium tuberculosis (MTB) invasion of A549 cell line (human lung adenocarcinoma cells). Small intefering RNA (siRNA) expressing vector targetting RhoA gene (si-RhoA) was constructed and transfered to A549 cells. MTB adhesion and invasion of A549 cells assays were performed 36 hours post transfection; The level of RhoA protein was determined by Western blot; The ultrastructural changes of cells were observed by electron microscope; Changes of filaments actin (F-actin) cytoskeleton were detected by laser scanning confocal microscope (LSCM)， and the number of MTB adhering to cell and internalized by cell were measured by quantitative polymerase chain reaction (PCR). These results showed that the level of RhoA protein decreased by 84.7% after A549 cells transfered by si-RhoA， with the capacity of MTB adhering to cell no change. The test of MTB invasion of cells showed that the index of F-actin rearrangement and the number of MTB internalized by cell were (63.0±3.1)% and (3.19±0.26)×103 copies for A549 cells transfered by si-RhoA， while they were (84.5±1.8)% and (4.45±0.29)×103 copies for A549 cells transfered by scramble siRNA， and the former was significantly less than the latter (P<0.01). All these results indicate that the protein of RhoA has a role on MTB invasion of A549 cells， which may mediate MTB invasion of non phagocytic cells by zipper mechanism.
CN
EN