CAJ | 학술논문

目的观察肿瘤坏死因子相关诱导凋亡配体(TRAIL)与丁酸纳(NaB)联合应用对肝癌细胞HepG-2的抑制作用.方法将HepG-2细胞悬液接种于96孔培养板后,分别或联合加入不同浓度的NaB(1.0、2.5、5.0 mmol/L)和TRAIL(10、100、500、1000μg/L),应用噻唑蓝(MTT)比色法检测其对HepG-2细胞生长抑制作用;流式细胞术检测细胞凋亡;逆转录-聚合酶链反应(RT-PCR)检测细胞内DR4和DR5 mRNA的表达水平.结果经不同浓度的TRAIL处理后,HepG-2细胞生长抑制率和凋亡率分别为(4.5±1.3)%和3.2±0.7)%,与对照组比较,差异无统计学意义(P>0.05).NaB对HepG-2细胞生长有明显抑制作用,且随着NaB浓度从1.0mmol/L升高到5.0mmol/L时,抑制率从(37.6±2.6)%升高到(58.4±3.0)%,各浓度组间差异有统计学意义(P<0.05),NaB浓度为2.5 mmol/L时的凋亡率为(26.7 5.2)%.联合应用NaB和TRAIL能够明显的提高HepG-2细胞的生长抑制率和凋亡率,与同等浓度的NaB或TRAIL单独应用比较,差异有统计学意义(P<0.05).HepG-2细胞经过NaB作用后,其DR5 mRNA的表达水平明显的高于对照组.结论 HepG-2细胞对单独应用TRAIL诱导的凋亡不敏感,NaB可通过上调HepG-2细胞内DR5 mRNA的表达而增强HepG-2对TRAIL的敏感性.
목적 관찰종류배사인자상관유도조망배체(TRAIL)여정산납(NaB)연합응용대간암세포HepG-2적억제작용.방법 장HepG-2세포현액접충우96공배양판후,분별혹연합가입불동농도적NaB(1.0、2.5、5.0 mmol/L)화TRAIL(10、100、500、1000μg/L),응용새서람(MTT)비색법검측기대HepG-2세포생장억제작용;류식세포술검측세포조망;역전록-취합매련반응(RT-PCR)검측세포내DR4화DR5 mRNA적표체수평.결과경불동농도적TRAIL처리후,HepG-2세포생장억제솔화조망솔분별위(4.5±1.3)%화3.2±0.7)%,여대조조비교,차이무통계학의의(P>0.05).NaB대HepG-2세포생장유명현억제작용,차수착NaB농도종1.0mmol/L승고도5.0mmol/L시,억제솔종(37.6±2.6)%승고도(58.4±3.0)%,각농도조간차이유통계학의의(P<0.05),NaB농도위2.5 mmol/L시적조망솔위(26.7 5.2)%.연합응용NaB화TRAIL능구명현적제고HepG-2세포적생장억제솔화조망솔,여동등농도적NaB혹TRAIL단독응용비교,차이유통계학의의(P<0.05).HepG-2세포경과NaB작용후,기DR5 mRNA적표체수평명현적고우대조조.결론 HepG-2세포대단독응용TRAIL유도적조망불민감,NaB가통과상조HepG-2세포내DR5 mRNA적표체이증강HepG-2대TRAIL적민감성.
Objective To investigate the inhibitory effects of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) combined with sodium butyrate (NaB) on hepatic carcinoma cell line HepG-2.Methods different concentrations of NaB( 1.0,2.5,5.0 mmol/L)and TRAIL ( 10,100,500,1000ng/ml) solely or combined with both added to the culture plates including HepG-2 suspension,inhibitory action of NaB and TRAIL on HepG-2 were Measured by MTT assays; The apoptotic rate of HepG-2 cells and the mRNA expressions of DR4 ,DR5 in HepG-2 cells were respectively detected by flow cytometry analysis and Real-time RT-PCR.Results By using different concentrations of TRAIL,the inhibitory rate of HepG-2 growth and HepG-2 apoptotic rate were ( 4.5 ± 1.3 ) % and 3.2 ± 0.7 ) % respectively,compared with control group,the differences were not significant statistically ( P ＞ 0.05 ).There was a dose-dependent inhibition of cell growth of HepG-2 by NaB ( P ＜ 0.05 ).The combination of the NaB and TRAIL can significantly increase the inhibitory rate of HepG-2 growth and HepG-2 apoptotic rate ( P ＜ 0.05 ).Expression level of DR5 mRNA of HepG-2 deal with NaB were higher than control group.Conclusion HepG-2 cells were resistant to apoptosis induced by TRAIL,NaB could enhance the sensitivity of Hep-2 cells to TRAIL by up-regulating the mRNA expressions of DR5 in HepG-2 cells.