Small-interference RNA targeting Survivin inhibits proliferation of human osteosarcoma cell line MG-63 in vitro and in vivo

Objective To explor the inhitory effect of small-interference RNA (siRNA) targeting Survivin on human osteosarcoma cell line MG-63 in vitro and in vivo. Methods A plasmid of a short hairpin RNA targeting Survivin was constructed, and transfected into MG-63 cell line by dosper liposomal method and selected by G418. Morphological change of tumor cells was observed under an inverted microscope. The expression of Survivin mRNA and protein was examined by using reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence microscopy respectively. The apoptotic cells were stained by acridine orange. The cell cycle was determined by flow cytometry. To establish the model of transplanted osteosarcoma in nude mice, the in vivo carcinogenic time, the weight and volume of subcutaneous tumors were measured. Results The number of drift cells was increased after transfection. Survivin shRNA could significantly reduce the expressions of Survivin mRNA and protein. The expression of Survivin mRNA after transfection was respectively inhibited by 85.08% with a PI value of 28. 20%. The apoptosis rate of the cells in the transfected group was 24. 54%. Flow cytometry analysis revealed an increase of the percentage of Go/G1 phase cells, along weth a decrease of cells population in the S and G2/M phase.After transferred by pSilence2. 1-neo-Survivin expression vector the carcinogenic activity of cancer cells was decreased, the carcinogenic time of the Survivin shRNA group and control group was 13. 2 ±2. 0 and 6. 5 ±1.6 days respectively (P ＜0. 01 ). The weigh and volume of subcutaneous tumors at the 4th week was inhibited by 59. 07% ( P ＜ 0. 01 ) and 62. 89% (P ＜ 0. 01 ) respectively. Conclusion Survivin shRNA could significantly reduce the expressions of Survivin mRNA and protein and inhibit the proliferation of the MG-63 cells, and increase the apoptosis of the cells. pSilence2. 1-neo-Survivin transfection could significantly inhibit the carcinogenic activity of the cells and the growth of the subcutaneous tumors in nude mice. Key words: Survivin;  RNA interference;  Osteosarcoma;  Nude mice;  Transplanted osteosarcoma