【摘要】 目的:探讨杭白菊提取液对肿瘤坏死因子相关凋亡诱导配体基因(TRAIL)抑制人大肠癌细胞株DLD-1作用的影响及其可能机制。方法:杭白菊提取液联合重组腺病毒载体(Ad)介导的TRAIL基因作用于人大肠癌细胞株DLD-1,通过倒置显微镜、MTT比色法和流式细胞仪,研究分析其对DLD-1细胞抑制作用的效果。采用逆转录聚合酶链反应(RT-PCR)和流式细胞术检测杭白菊提取液作用前后DLD-1细胞TRAIL、TRAIL受体(TRAIL-Rs)mRNA以及细胞表面TRAIL蛋白表达的变化。结果:Ad/hTERT-gTRAIL对DLD-1细胞的生长抑制率和凋亡率分别为31·4%和13·5%;联合杭白菊提取液后,生长抑制率和凋亡率均显著提高,达93·1%和45·4%(P<0·05)。杭白菊提取液作用后DLD-1细胞TRAIL mRNA的表达量从作用前的0·46上调至1·01,细胞表面TRAIL蛋白表达的百分率从作用前的2·2%升高到5·0%(P<0·05)。TRAIL死亡受体(DR4、DR5)mRNA的表达量分别从0·70和0·22上调至1·10和0·83(P<0·05),而TRAIL诱骗受体(DcR1、DcR2)mRNA的表达量从0·60和1·15下调至0·19和0·78(P<0·05)。结论:杭白菊提取液联合重组腺病毒载体(Ad)介导的TRAIL基因(Ad/hTERT-gTRAIL)能有效诱导DLD-1细胞的凋亡。杭白菊提取液上调TRAIL死亡受体表达以及下调TRAIL诱骗受体的表达可能在增强TRAIL诱导的凋亡作用中起着重要作用。更多还原

【Abstract】 AIM: To explore the influence of Chrysanthemum morifolium Ramat on TNF-related apoptosis inducing ligand (TRAIL)-mediated apoptosis in human colon cancer cell line DLD-1 and its possible mechanism. METHODS: Adenovirus-mediated TRAIL gene AD/hTERT-gTRAIL was applied either alone or by combination with Chrysanthemum morifolium Ramat in human colon DLD-1 cell line. Cell growth and apoptosis were measured by inverted microscope, MTT method and flow cytometry. The expression of TRAIL mRNA, TRAIL-Rs mRNA and TRAIL protein expression after exposure to Chrysanthemum morifolium Ramat were measured by semi-quantitive RT-PCR and FACS, respectively. RESULTS: The suppression percentages and apoptotic rate of DLD-1 by Ad/hTERT-gTRAIL alone were 31.4% and 13.5%, respectively. Combination of TRAIL gene transfection with Chrysanthemum morifolium Ramat, the suppression and the apoptosis rate raised to 93.1% and 45.4%, respectively (P<0.05). The expression of TRAIL mRNA was upregulated from 0.46 to 1.01 by Chrysanthemum morifolium Ramat (P<0.05). The percentage of TRAIL protein expression increased from 2.2% to 5.0%. DR4 mRNA and DR5 mRNA were upregulated from 0.70, 0.22 to 1.10, 0.83, respectively (P<0.01), while DcR1 mRNA, DcR2 mRNA were downregulated from 1.60, 1.15 to 0.19, 0.78, respectively (P<0.05). CONCLUSIONS: Ad/hTERT-gTRAIL induced apoptosis in DLD-1.Combination of Chrysanthemum morifolium Ramat with Ad/hTERT-gTRAIL enhanced the effect. The upregulation of TRAIL death receptors as well as the downregulation of TRAIL decoy receptors may play an important role.更多还原