CAJ | 학술논문

目的通过采用RNA干扰技术下调survivin基因在人肝癌SMMC-7721细胞中的表达.观察survivin表达下调对细胞G2/M期阻滞、细胞凋亡和重离子辐射敏感性的影响.方法针对survivin mRNA体外化学合成小干扰RNA(siRNA),转染细胞,以实时PCR方法检测转染后24和48 h细胞survivin mRNA的表达.Annexin-FITC检测细胞的凋亡情况;流式细胞仪检测周期阻滞情况;克隆存活法确定细胞的辐射敏感性.结果转染24和48 h后,细胞中survivin的表达量分别为未转染组的59%和39%;转染survivin siRNA后24 h,引起细胞G2/M期阻滞,48 h阻滞明显.转染survivin siRNA 48 h后的细胞凋亡率为21.41%,明显高于未转染组细胞(t=9.13,P<0.01).siRNA转染组细胞受辐照后的存活率明显降低.结论以siRNA下调survivin的表达可有效诱导细胞凋亡,引起G2/M期阻滞,提高细胞对重离子的辐射敏感性.
목적 통과채용RNA간우기술하조survivin기인재인간암SMMC-7721세포중적표체.관찰survivin표체하조대세포G2/M기조체、세포조망화중리자복사민감성적영향.방법 침대survivin mRNA체외화학합성소간우RNA(siRNA),전염세포,이실시PCR방법검측전염후24화48 h세포survivin mRNA적표체.Annexin-FITC검측세포적조망정황;류식세포의검측주기조체정황;극륭존활법학정세포적복사민감성.결과 전염24화48 h후,세포중survivin적표체량분별위미전염조적59%화39%;전염survivin siRNA후24 h,인기세포G2/M기조체,48 h조체명현.전염survivin siRNA 48 h후적세포조망솔위21.41%,명현고우미전염조세포(t=9.13,P<0.01).siRNA전염조세포수복조후적존활솔명현강저.결론 이siRNA하조survivin적표체가유효유도세포조망,인기G2/M기조체,제고세포대중리자적복사민감성.
Objective To investigate the influences of survivin down-regulation on cell G2/M phase arrest,apeptosis and sensitivity to carbon ion irradiation. Methods Small interfering RNA (siRNA) targeting survivin mRNA was designed, in vitro chemo-synthesized and transfected into SMMC-7721 cells. Survivin mRNA expression in SMMC-7721 cells was measured by real-time PCR, and the apeptotic rates by Annexin-FTTC at 24 and 48 h after transfection. Cell G2/M phase arrest after transfection was assessed with flow eytometry as well. Cellular sensitivity to high-LET carbon ions was determined by means of colony-forming assay. Results The expressions of survivin at mRNA level were down-regulated to be 59% and 39% in relation to the non-treated cells at 24 and 48 h after siRNA transfeetion, respectively. G2/M phase arrest in SMMC-7721 cells at 24 h after transfection was observed while much more obvious at 48 h. The apeptotic rate of SMMC-7721 cells was 21.41 % at48 h after survivin siRNA transfection, which was significantly higher than that of the cells transfected with negative siRNA. Moreover, a decreased clonogenic survival in siRNA treated group was shown. Conclusion Down-regulation of survivin gene expression in SMMC-7721 cells by siRNA could effectively induce cell apeptosis and G2/M phase arrest, and enhance the cellular radiosensitivity to high-LET heavy ions.