1.Effects of exogenous VEGF(165)b on invasion and migration of human lung adenocarcinoma A549 cells.
Jing, CHEN ; Zhenyu LI ; Sheng, ZHANG ; Ruiguang ZHANG ; Meera DASSARATH ; Gang, WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(5):619-24
Vascular endothelial growth factor 165 (VEGF(165))-mediated autocrine stimulation of tumor cells enhances the progression to a malignant phenotype. VEGF(165)b competes with VEGF(165) and binds to vascular endothelial growth factor receptor (VEGFR), resulting in inhibition of downstream signal transduction pathways. This study was designed to investigate the role of VEGF(165)b in the migration and invasion of human lung adenocarcinoma A549 cells. The full-length of VEGF(165)b was constructed and cloned into an expression plasmid (pVEGF(165)b), and then transfected into A549 cells. Dimethylthiazolyl- 1 -2, 5-diphenyltetrazolium bromide (MTT) assay was used to detect the effect of VEGF(165)b on proliferation of transfected cells. Reverse transcription polymerase chain reaction (RT-PCR) was employed to examine the effect of VEGF(165)b on the expression of VEGF(165) in transfected cells. Wound-healing assays were used to investigate the effect of VEGF(165)b on migration of transfected cells. Matrix metalloproteinase (MMPs) activity assay and in vitro invasion assay were used to determine the role of VEGF(165)b in invasion of transfected cells. There was no significant change in proliferation of A549 cells after transfection of pVEGF(165)b, but the expression of VEGF(165), migration and invasion in A549 cells were inhibited. Furthermore, exogenous VEGF(165)b inhibited the activity of MMP9 in the supernatant of A549 cells and the subsequent invasion capacity of those cells. We therefore conclude that exogenous VEGF(165)b can inhibit the expression of VEGF(165), as well as the migration and invasion of A549 cells, but has no effect on the proliferation of A549 cells.
2.Contribution of decreased expression of Ku70 to enhanced radiosensitivity by sodium butyrate in glioblastoma cell line (U251).
Yuhui, LI ; Hongxia, ZHOU ; Enming, XING ; Meera, DASSARATH ; Jinghua, REN ; Xiaorong, DONG ; Hongli, LIU ; Kunyu, YANG ; Gang, WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(3):359-64
The present study investigated the enhanced radiosensitivity of U-251 cells induced by sodium butyrate (NaB) and its possible mechanisms. Increased radiosensitivity of U251 cells was examined by clonogenic cell survival assays. The expression of Ku70 mRNA and protein was detected by using RT-PCR and Western blotting respectively. γ-H2AX foci were measured at different time points after ionizing irradiation alone or combined with NaB treatment. The results showed that cell survival rate was significantly reduced, both D0 and Dq values were decreased (D0: 1.43 Gy vs. 1.76 Gy; Dq: 1.22 Gy vs. 2.05 Gy) after the combined treatment as compared with irradiation alone, and sensitivity enhancing ratio (SER) reached 1.23. The average number of γ-H2AX foci per cell receiving the combined treatment was significantly increased at different time points, and the expression levels of Ku70 mRNA and protein were suppressed by NaB in a dose-dependent manner. It was concluded that enhanced radiosensitivity induced by NaB involves an inhibited expression of Ku70 and an increase in γ-H2AX foci, which suggests decreased ability in DSB repair.
3.Effects of Exogenous VEGF165b on Invasion and Migration of Human Lung Adenocarcinoma A549 Cells
CHEN JING ; LI ZHENYU ; ZHANG SHENG ; ZHANG RUIGUANG ; Dassarath MEERA ; WU GANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(5):619-624
Vascular endothelial growth factor 165 (VEGF165)-mediated autocrine stimulation of tumor cells enhances the progression to a malignant phenotype.VEGF165b competes with VEGF165 and binds to vascular endothelial growth factor receptor (VEGFR),resulting in inhibition of downstream signal transduction pathways.This study was designed to investigate the role of VEGF165b in the migration and invasion of human lung adenocarcinoma A549 cells.The full-length of VEGF165b was constructed and cloned into an expression plasmid (pVEGF165b),and then transfected into A549 cells.Dimethylthiazolyl-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect the effect of VEGF165b on proliferation of transfected cells.Reverse transcription polymerase chain reaction (RT-PCR) was employed to examine the effect of VEGF165b on the expression of VEGF165 in transfected cells.Wound-healing assays were used to investigate the effect of VEGF165b on migration of transfected cells.Matrix metalloproteinase (MMPs) activity assay and in vitro invasion assay were used to determine the role of VEGF165b in invasion of transfected cells.There was no significant change in proliferation of A549 cells after transfection of pVEGF165b,but the expression of VEGF165,migration and invasion in A549 cells were inhibited.Furthermore,exogenous VEGF165b inhibited the activity of MMP9 in the supematant of A549 cells and the subsequent invasion capacity of those cells.We therefore conclude that exogenous VEGF165b can inhibit the expression of VEGF165,as well as the migration and invasion ofA549 cells,but has no effect on the proliferation ofA549 cells.
4.Contribution of Decreased Expression of Ku70 to Enhanced Radiosensitivity by Sodium Butyrate in Glioblastoma Cell Line (U251)
LI YUHUI ; ZHOU HONGXIA ; XING ENMING ; Dassarath MEERA ; REN JINGHUA ; DONG XIAORONG ; LIU HONGLI ; YANG KUNYU ; WU GANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(3):359-364
The present study investigated the enhanced radiosensitivity of U-251 cells induced by sodium butyrate (NaB) and its possible mechanisms.Increased radiosensitivity of U251 cells was examined by clonogenic cell survival assays.The expression of Ku70 mRNA and protein was detected by using RT-PCR and Western blotting respectively.γ-H2AX foci were measured at different time points after ionizing irradiation alone or combined with NaB treatment.The results showed that cell survival rate was significantly reduced,both D0 and Dq values were decreased (D0:1.43 Gy vs.1.76 Gy; Dq:1.22 Gy vs.2.05 Gy) after the combined treatment as compared with irradiation alone,and sensitivity enhancing ratio (SER) reached 1.23.The average number ofγ-H2AX foci per cell receiving the combined treatment was significantly increased at different time points,and the expression levels of Ku70mRNA and protein were suppressed by NaB in a dose-dependent manner.It was concluded that enhanced radiosensitivity induced by NaB involves an inhibited expression of Ku70 and an increase in γ-H2AX foci,which suggests decreased ability in DSB repair.