ABSTRACT
Objective:To evaluate the effect of Salvianolic acid B on the radiosensitivity of human non-small cell lung cancer cells and investigate its possible mechanism.Methods:Non-small cell lung cancer cells A549 and H1299 were cultured in vitro. The toxicity of Salvianolic acid B on non-small cell lung cancer cells was detected by MTT assay. The effect of Salvianolic acid B on the radiosensitivity was assessed by clone formation assay. Transwell chamber assay was used to evaluate the effect of Salvianolic acid B on the migration of tumor cells. Western blot was employed to detect the expression levels of OTUD7B, MMP-2, MMP-9, E-cadherin, Akt and p-Akt regulated by Salvianolic acid B. Results:Salvianolic acid B (5 μmol/L) could inhibit the proliferation of A549 and H1299 cells. Clone formation assay showed that Salvianolic acid B increased the radiosensitivity of A549 and H1299 cells, with a radiosensitization ratio of 1.45 and 1.38, respectively. Transwell chamber assay indicated that the ability of cell migration was significantly inhibited by Salvianolic acid B ( P<0.05). Western blot revealed that the expression levels of OTUD7B in A549 and H1299 cells were induced by irradiation in a time-dependent manner. Salvianolic acid B could down-regulate the expression levels of MMP-2, MMP-9 and p-Akt, whereas up-regulate the expression level of E-cadherin by down-regulating the expression level of OTUD7B. Conclusions:Salvianolic acid B can enhance the radiosensitivity of A549 and H1299 cells. The possible mechanism is that Salvianolic acid B down-regulates the expression level of OTUD7B induced by irradiation and inhibits the epithelial-mesenchymal transition process of tumor cells.
ABSTRACT
In the interaction between wheat and Puccinia triticina, wheat resists the invasion of P. triticina by producing hypersensitive reaction-programmed cell death (HR-PCD). To better understand the mechanism of wheat resistance to P. triticina, it is important to identify the defensive genes involved in wheat resistance to leaf rust. This paper systematically presented the role of Wrab17 in the HR-PCD process in wheat after infection by P. triticina. The subcellular location analysis is performed using the full length of Wrab17 coding sequences and Wrab17 is found to be localized in cell nucleus and cytoplasm. Quantitative RT-PCR (RT-qPCR) and western blot analysis showed that expression of Wrab17â¯at both mRNA and protein levels increased by P. triticina infection, indicating that the Wrab17 gene participated in the interaction between wheat and P. triticina. Wrab17 knockdown plants were generated by RNA-mediated gene silencing technology (RNA interference, RNAi) and confirmed by southern blot. Further analysis with P. triticina inoculation found that knockdown of Wrab17 exhibited decreased HR expansion area in incompatible combination (L10×260) and significant higher sensitivity to the compatible pathogen P. triticina race 165. In all, this work reports that Wrab17 is a new defensive gene playing a role in wheat resistance to leaf rust.
