CAJ | 학술논문

目的探讨RNA干扰技术沉默Ras homolgyC(RhoC)基因对卵巢癌细胞SKOV3细胞对紫杉醇敏感性的影响.方法体外设计合成针对RhoC基因的微小干扰RNA(miRNA),转染卵巢癌细胞SKOV3,应用RT-PCR和Western印迹方法检测转染前后基因mRNA和蛋白的变化,应用MTT方法检测干扰后SKOV3细胞对紫杉醇敏感性的变化,同时应用RT-PCR法检测转染前后SKOV3细胞中的凋亡基因Caspase-3、Survivin mRNA的表达水平.结果转染针对RhoC基因的miRNA后24、48、72 h,SKOV3细胞mRNA和蛋白水平显著下降;关闭RhoC基因后,SKOV3细胞对紫杉醇的敏感性显著提高.与单独使用紫杉醇相比,在RhoC-miRNA联合紫杉醇组中促进凋亡的基因Caspase-3明显增高,而抗凋亡蛋白Suvlvin mRNA水平显著降低.结论 RNA干扰RhoC基因能增加卵巢癌SKOV3细胞对紫杉醇的敏感性,其协同机制可能与Caspase-3基因的上调,Suvivin基因的下调有关.
목적 탐토RNA간우기술침묵Ras homolgyC(RhoC)기인대란소암세포SKOV3세포대자삼순민감성적영향.방법 체외설계합성침대RhoC기인적미소간우RNA(miRNA),전염란소암세포SKOV3,응용RT-PCR화Western인적방법검측전염전후기인mRNA화단백적변화,응용MTT방법검측간우후SKOV3세포대자삼순민감성적변화,동시응용RT-PCR법검측전염전후SKOV3세포중적조망기인Caspase-3、Survivin mRNA적표체수평.결과 전염침대RhoC기인적miRNA후24、48、72 h,SKOV3세포mRNA화단백수평현저하강;관폐RhoC기인후,SKOV3세포대자삼순적민감성현저제고.여단독사용자삼순상비,재RhoC-miRNA연합자삼순조중촉진조망적기인Caspase-3명현증고,이항조망단백Suvlvin mRNA수평현저강저.결론 RNA간우RhoC기인능증가란소암SKOV3세포대자삼순적민감성,기협동궤제가능여Caspase-3기인적상조,Suvivin기인적하조유관.
Objective To explore the changes of paclitaxel sensitivity by RNA interference of RhoC gene in ovarian cancer cell lines. Methods The microRNA (miRNA) targeting RhoC gene was designed and synthesized by in vitro transcription and transfected into ovarian cancer cell line SKOV3. The mRNA and protein of RhoC were detected by the means of RT-PCR and Western blot. The changes of paclitaxel sensitivity after interference were examined by methyl thiazolyl tetrasolium (MTT) assay. At the same time, the mRNA of apoptosis gene Caspase-3 and survivin were evaluated by RT-PCR. Results In SKOV3 cell, the mRNA and protein of RhoC dramatically decreased at 24, 48 and 72 hours post-transfection. The sensitivity of SKOV3 cell line to paclitaxei increased significantly after turning off the RhoC gene. The pro-apoptotic gene Caspase-3 increased significantly and anti-apoptosis gene survivin dramatically decreased after the combination treatment of paclitaxel and RhoC-miRNA in comparison with paclitaxel alone treatment. Conclusion The sensitivity of ovarian cancer cell lines SKOV3 to paclitaxel can be enhanced by RNA interfering RhoC gene. The synergistic mechanism is possibly correlated with the up-regulation of Caspase-3 gene and the down-regulation of suvivin gene.