Synergistic anti-osteosarcoma effects of TRAIL overexpression in combination with Ether à go-go 1 silence
ZENG Wen-rong
CHEN Zhi-da
LIU Qing-jun
LIN Bin
WU Xin-yu
WU Jin
Abstract：Objective To evaluate effects of TNF-related apoptosis-inducing ligand ( TRAIL ) overexpression in combination with Ether à go-go 1 ( Eag 1 ) silence on osteosarcoma. Methods Several adenoviral vectors named Ad5. eGFP, CRAd5. siEag, CRAd5. TRAIL and CRAd5. TRAIL / siEag1 were generated and their anti-tumor effects on osteosarcoma were investigated. Results Ad5. eGFP had no effects on the expression of Eag 1 and TRAIL in MG-63 cells, while CRAd5. siEag knocked down the expression of Eag 1, CRAd5. TRAIL over expressed the TRAIL expression and CRAd5. TRAIL / siEag1 permitted simultaneous overexpression of TRAIL and knockdown of Eag 1.Results of cell proliferation showed that CRAd5. TRAIL / siEag1 induced growth arrest in a more efficient manner than CRAd5. TRAIL or CRAd5. siEag1, and had no effects on human osteoblastic hFOB 1.19 cells. The amount of activated Caspase-3 / 7 was ( 39693.33 ± 1922.96 ), ( 2832.83 ± 375.99 ), ( 3242.50 ± 329.97 ) and ( 15583.17 ± 698.54 ) RLU in CRAd5. TRAIL / siEag1, Ad5. eGFP, CRAd5. siEag and CRAd5. TRAIL group respectively. Results showed that the amount of activated Caspase-3 / 7 was significantly higher in CRAd5. TRAIL / siEag1 infected cells than in other adenoviral vectors infected cells ( P < 0.001 ). Moreover, the amount of cleaved PARP was also significantly higher in CRAd5. TRAIL / siEag1 infected cells than in other adenoviral vectors infected cells. At 8 day after local injection, the tumor volume in CRAd5. TRAIL / siEag1 was ( 231.19 ± 25.96 ) mm3, which was significantly smaller than the volumes in Ad5. eGFP ( 504.68 ± 102.32 ) mm3, CRAd5. siEag ( 365.85 ± 47.60 ) mm3 or CRAd5. TRAIL ( 355.48 ± 35.34 ) mm3 group ( P < 0.05 ). At 10 th, 12 th and 14 th day after local injection, the tumor volumes in CRAd5. TRAIL / siEag1 group were ( 367.82 ± 39.52 ), ( 470.18 ± 60.11 ) and ( 631.52 ± 61.22 ) mm3, while the tumor volumes in Ad5. eGFP group were ( 826.57 ± 130.60 ), ( 1284.69 ± 243.50 ) and ( 1762.06 ± 180.36 ) mm3, in CRAd5. siEag group were ( 552.12 ± 64.39 ), ( 721.82 ± 108.20 ) and ( 949.20 ± 124.70 ) mm3 and in CRAd5. TRAIL group were ( 524.03 ± 43.47 ), ( 653.54 ± 56.22 ) and ( 896.44 ± 93.64 ) mm3. Results showed that the tumor volume was significantly smaller in CRAd5. TRAIL / siEag1 injected animals compared with Ad5. eGFP, CRAd5. siEag1 or CRAd5. TRAIL injected animals. The tumors from the mice injected with CRAd5. TRAIL / siEag1 demonstrated extensive apoptosis ( 27.33 ± 2.07 ) %, compared with the group treated with Ad5. eGFP, CRAd5. siEag1 or CRAd5. TRAIL. The apoptotic index were ( 2.68 ± 0.64 ) %, ( 8.49 ± 1.06 ) % and ( 15.99 ± 1.94 ) % in Ad5. eGFP, CRAd5. siEag1 and CRAd5. TRAIL, respectively. The differences were significant ( P < 0.001 ). Conclusions CRAd5. TRAIL /siEag1 can represent an effective strategy for osteosarcoma gene therapy due to the synergistic anti-tumor effects of Eag 1 knockdown and TRAIL overexpression.
Keywords：OsteosarcomaCell proliferationApoptosisGenesTumor necrosis factor-related apoptosis-inducing ligandEther à go-go1
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 123-128 )