[Inhibitory effect of multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes on human colon cancer cells proliferation].
In order to maximize the efficiency and versatility of the vector-based siRNA approach, we have developed a novel siRNA expression vector containing multiple tandem siRNA cassettes to investigate the synergistic inhibitory effect of it on human colon cancer cells proliferation. Multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed and transfected into human colon cancer cell. The effect of multiple siRNA recombinant expression vector was detected by RT-PCR, Western blotting and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes was constructed successfully. Livin and Survivin genes inhibition ratio of Livin and Survivin siRNA at mRNA levels were 27.90% and 32.24%, at protein levels were 22.28% and 40.86%, the apoptotic ratio was (11.69 +/- 1.37) %, but the synergistic effect was weaker than Livin and Survivin RNA interference, respectively. The multiple siRNA recombinant expression vector targeting simultaneously Livin and Survivin genes has been constructed successfully. It can inhibit the expression of Livin and Survivin genes in human colon cancer cells, but the synergistic effect was weaker than Livin and Survivin RNA interferences, respectively.
Cai
Ming
M
Department of General Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Wang
Guo-Bin
GB
Tao
Kai-Xiong
KX
Cai
Chang-Xue
CX
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D000077022:Survivin; D014162:Transfection
D048868:Adaptor Proteins, Signal Transducing; C107489:BIRC5 protein, human; C419866:BIRC7 protein, human; D051034:Inhibitor of Apoptosis Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D000077022:Survivin