陕西医学杂志
陝西醫學雜誌
협서의학잡지
Shaanxi Medical Journal
2015年
11期
1446-1449
,共4页
RNA 干扰%视网膜母细胞瘤%多药耐药相关蛋白质类%P-糖蛋白%药物耐受性
RNA 榦擾%視網膜母細胞瘤%多藥耐藥相關蛋白質類%P-糖蛋白%藥物耐受性
RNA 간우%시망막모세포류%다약내약상관단백질류%P-당단백%약물내수성
RNA interference%Retinoblastoma%Multidrug resistance-Associated proteines%P-Glyco-protein%Drug tolerance
目的：探讨应用 RNA 干扰(RNAi)技术逆转视网膜母细胞瘤多药耐药的可行性。方法：将设计合成的针对多药耐药基因 MDR1的特异性小分子干扰 RNA(siRNA),用脂质体转染具有 MDR1基因高表达的视网膜母细胞瘤耐药细胞株 SO-Rb50/VCR。用实时定量 RT-PCR 技术和 Westernblot 分别测定转染前后细胞 MDR1 mRNA 及 P-糖蛋白(P-gp)表达的变化；用 CCK-8法和 TUNEL 法检测转染前细胞株(VCR 组),转染后细胞(VCR+siRNA 组)对不同浓度长春新碱、依托泊苷和卡铂药物敏感性。结果：VCR+siRNA 组较 VCR 组比较,MDR1基因在 mRNA 水平和蛋白水平表达显著下降,差异具有统计学意义。随着长春新碱、依托泊苷浓度的升高 VCR+siR-NA 组较 VCR 组比较,细胞存活率显著降低,差异具有统计学意义。不同浓度的卡铂作用于 VCR组、CT 组和 VCR+siRNA 组细胞存活率、细胞凋亡率各组见未见显著性差异。随着长春新碱、依托泊苷浓度的升高,VCR+siRNA 组较 VCR 组比较,细胞凋亡率显著性升高,差异具有统计学意义。结论：在 SO-Rb50/VCR 细胞中,针对 MDR1合成的 siRNA 能够有效地抑制 MDR1 mRNA 和P-gp 的表达,并能恢复肿瘤细胞对长春新碱和依托泊苷的敏感性。应用 RNAi 技术,能够逆转 P-gp 引起的视网膜母细胞瘤多药耐药性。
目的：探討應用 RNA 榦擾(RNAi)技術逆轉視網膜母細胞瘤多藥耐藥的可行性。方法：將設計閤成的針對多藥耐藥基因 MDR1的特異性小分子榦擾 RNA(siRNA),用脂質體轉染具有 MDR1基因高錶達的視網膜母細胞瘤耐藥細胞株 SO-Rb50/VCR。用實時定量 RT-PCR 技術和 Westernblot 分彆測定轉染前後細胞 MDR1 mRNA 及 P-糖蛋白(P-gp)錶達的變化；用 CCK-8法和 TUNEL 法檢測轉染前細胞株(VCR 組),轉染後細胞(VCR+siRNA 組)對不同濃度長春新堿、依託泊苷和卡鉑藥物敏感性。結果：VCR+siRNA 組較 VCR 組比較,MDR1基因在 mRNA 水平和蛋白水平錶達顯著下降,差異具有統計學意義。隨著長春新堿、依託泊苷濃度的升高 VCR+siR-NA 組較 VCR 組比較,細胞存活率顯著降低,差異具有統計學意義。不同濃度的卡鉑作用于 VCR組、CT 組和 VCR+siRNA 組細胞存活率、細胞凋亡率各組見未見顯著性差異。隨著長春新堿、依託泊苷濃度的升高,VCR+siRNA 組較 VCR 組比較,細胞凋亡率顯著性升高,差異具有統計學意義。結論：在 SO-Rb50/VCR 細胞中,針對 MDR1閤成的 siRNA 能夠有效地抑製 MDR1 mRNA 和P-gp 的錶達,併能恢複腫瘤細胞對長春新堿和依託泊苷的敏感性。應用 RNAi 技術,能夠逆轉 P-gp 引起的視網膜母細胞瘤多藥耐藥性。
목적：탐토응용 RNA 간우(RNAi)기술역전시망막모세포류다약내약적가행성。방법：장설계합성적침대다약내약기인 MDR1적특이성소분자간우 RNA(siRNA),용지질체전염구유 MDR1기인고표체적시망막모세포류내약세포주 SO-Rb50/VCR。용실시정량 RT-PCR 기술화 Westernblot 분별측정전염전후세포 MDR1 mRNA 급 P-당단백(P-gp)표체적변화；용 CCK-8법화 TUNEL 법검측전염전세포주(VCR 조),전염후세포(VCR+siRNA 조)대불동농도장춘신감、의탁박감화잡박약물민감성。결과：VCR+siRNA 조교 VCR 조비교,MDR1기인재 mRNA 수평화단백수평표체현저하강,차이구유통계학의의。수착장춘신감、의탁박감농도적승고 VCR+siR-NA 조교 VCR 조비교,세포존활솔현저강저,차이구유통계학의의。불동농도적잡박작용우 VCR조、CT 조화 VCR+siRNA 조세포존활솔、세포조망솔각조견미견현저성차이。수착장춘신감、의탁박감농도적승고,VCR+siRNA 조교 VCR 조비교,세포조망솔현저성승고,차이구유통계학의의。결론：재 SO-Rb50/VCR 세포중,침대 MDR1합성적 siRNA 능구유효지억제 MDR1 mRNA 화P-gp 적표체,병능회복종류세포대장춘신감화의탁박감적민감성。응용 RNAi 기술,능구역전 P-gp 인기적시망막모세포류다약내약성。
Objective :To investigate the effects of small interference RNA(siRNA)on the inhibition of MDR1 mRNA and reversal of drug resistance.Methods:The retinoblastoma (RB)multidrug resistant cells (SO-Rb50/VCR)were transfected with human MDR1 specific double-stranded,small interfering siRNA molecules.The expression of MDR1 mRNA was measured by realtime PCR and p-gp expression was detected by westernblot.Cell survival rate of SO-Rb50/VCR before(VCR group)and after transfection(VCR+siRNA group)was determined u-sing a standard colorimetric cell counting kit-8 (CCK-8)to check the drug the sensitivity to three different chemo-therapeutic.Rb cells apoptosis was analyzed by TUNEL assay.Result:The protein and mRNA expression level of MDR1 decreased significantly in the VCR+siRNA group compared to that in the VCR group.The percentage of cell survival decreased in VCR+siRNA group compared to that in the VCR group with the high dose of Vincristine or et-oposide.The percentage of apoptotic cells to total cells was significantly higher in the siRNA+VCR group than that in the VCR group with the high dose of Vincristine or etoposide.There was no significant difference between groups when RB cells were treated by various dose of carboplatin.Conclusions:Inhibited the express of MDR1 mRNA and p-gp by siRNA ,SO-Rb50/VCR cells were sensitive to vincristine and etoposide.RNAi can reverse the drug resistance to chemotherapeutic agents which were transferred by P-gp.
