CAJ | 학술논문

目的观察荷载肿瘤坏死因子相关诱导凋亡配体(TRAIL)基因复制缺陷型腺病毒(Ad-TRAIL)联合表柔比星(EPB)对膀胱癌T24细胞增殖和凋亡的影响.方法将T24细胞分为Ad-TRAIL联合EPB组、Ad-TRAIL组、EPB组和PBS组.干预细胞后,应用结晶紫法检测细胞毒性;Western blot法检测TRAIL蛋白表达;细胞计数试剂盒(CCK-8)法检测细胞增殖;膜联蛋白V(Annexin V)-异硫氰酸荧光素(FITC)和原位末端转移酶标记(TUNEL)法检测细胞凋亡.结果结晶紫结果表明Ad-TRAIL联合EPB较单一等剂量Ad-TRAIL、EPB对T24细胞毒性明显;Western blot结果表明Ad-TRAIL联合EPB作用的T24细胞仍高效表达TRAIL;CCK-8结果表明10.0病毒滴度(MOI)的Ad-TRAIL联合2.0 mg/L的EPB干预4d后细胞抑制率为(56.8±3.5)％,与单一等剂量Ad-TRAIL、EPB比较,差异有统计学意义(P＜0.01).TUNEL结果表明10.0 MOI的Ad-TRAIL联合2.0 mg/L的EPB干预48 h后凋亡率为(56.4±3.6)％,与对应剂量的Ad-TRAIL (34.1±3.3)％、EPB(21.3 ±2.2)％比较,差异有统计学意义(P＜0.05);Annexin V-FITC显示10.0 MOI的Ad-TRAIL联合2.0 mg/L的EPB干预48 h后凋亡细胞数分别为Ad-TRAIL、EPB的2.2、2.0倍,差异有统计学意义(P＜0.05).结论 Ad-TRAIL联合EPB有明显的抑制T24细胞增殖和促进凋亡的作用,同时具有显著协同效果.
목적 관찰하재종류배사인자상관유도조망배체(TRAIL)기인복제결함형선병독(Ad-TRAIL)연합표유비성(EPB)대방광암T24세포증식화조망적영향.방법 장T24세포분위Ad-TRAIL연합EPB조、Ad-TRAIL조、EPB조화PBS조.간예세포후,응용결정자법검측세포독성;Western blot법검측TRAIL단백표체;세포계수시제합(CCK-8)법검측세포증식;막련단백V(Annexin V)-이류청산형광소(FITC)화원위말단전이매표기(TUNEL)법검측세포조망.결과 결정자결과표명Ad-TRAIL연합EPB교단일등제량Ad-TRAIL、EPB대T24세포독성명현;Western blot결과표명Ad-TRAIL연합EPB작용적T24세포잉고효표체TRAIL;CCK-8결과표명10.0병독적도(MOI)적Ad-TRAIL연합2.0 mg/L적EPB간예4d후세포억제솔위(56.8±3.5)％,여단일등제량Ad-TRAIL、EPB비교,차이유통계학의의(P＜0.01).TUNEL결과표명10.0 MOI적Ad-TRAIL연합2.0 mg/L적EPB간예48 h후조망솔위(56.4±3.6)％,여대응제량적Ad-TRAIL (34.1±3.3)％、EPB(21.3 ±2.2)％비교,차이유통계학의의(P＜0.05);Annexin V-FITC현시10.0 MOI적Ad-TRAIL연합2.0 mg/L적EPB간예48 h후조망세포수분별위Ad-TRAIL、EPB적2.2、2.0배,차이유통계학의의(P＜0.05).결론 Ad-TRAIL연합EPB유명현적억제T24세포증식화촉진조망적작용,동시구유현저협동효과.
Objective To observe the effects of the combined use of Ad-tumor necrosis factor-related apoptosis-inducing ligand (Ad-TRAIL) with Epirubicin on proliferation and apoptosis of bladder cancer cell line T24.Methods T24 cells were divided into 4 groups:Ad-TRAIL plus EPB group,Ad-TRAIL group,EPB group and PBS group.Cytotoxic effect was determined by using violet crystal staining method.TRAIL protein was detected by using Western blotting.The inhibitory rate of cells was assessed by using cell counting kit-8 (CCK-8) assay.Cell apoptosis was analyzed by AnnexinV and TdT-mediated dUTP nick end labeling (TUNEL) assay,respectively.Results Violet Crystal staining method indicated the cytotoxicity to T24 cells in Ad-TRAIL plus EPB group was significantly increased as compared with Ad-TRAIL group and EPB group.Western blotting confirmed that T24 cells treated with Ad-TRAIL plus EPB could express TRAIL with high efficiecy.CCK-8 assay revealed the inhibitory rate after 48 h in Ad-TRAIL plus EPB group [(56.8 ± 3.5) ％] was significandy higher than in Ad-TRAIL group and EPB group (P ＜0.01).TUNEL demonstrated that the apoptosis rate after 48 h in Ad-TRAIL plus EPB group [(53.4 ±3.6)％] was significantly higher than in Ad-TRAIL group [(34.1 ±3.3)％] and EPB group [(21.3 ±2.2) ％].Conclusion The combined use of Ad-TRAIL with EPB could increase cytotoxic activity and apoptosis rate synergistically,which may provide novel strategies for the treatment of bladder cancer.