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山东大学学报(医学版)

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RAIL泰素帝协同诱导人肺腺癌细胞凋亡的研究

周伟   

  1. 1.山东大学齐鲁医院肿瘤防治中心,山东 济南 250012;
  • 收稿日期:2005-09-21 修回日期:1900-01-01 出版日期:2006-05-24 发布日期:2006-05-24
  • 通讯作者: 周伟

Docetaxel sensitizes human lung adenocarcinoma cellsto TRAILinduced apoptosis

ZHOU Wei   

  1. 1. Cancer Center, Qilu Hospital of Shandong University, Jinan 250012, Shandong, China;
  • Received:2005-09-21 Revised:1900-01-01 Online:2006-05-24 Published:2006-05-24
  • Contact: ZHOU Wei

摘要: 目的:探讨肿瘤坏死因子相关的凋亡诱导配体(tumor necrosis factor related apoptosis inducing ligand,TRAIL)和化疗药物泰素帝(TXT)联合应用对体外培养人肺腺癌A549细胞的凋亡诱导作用,并初步探讨其作用机制。方法:通过形态学观察和MTT比色法检测TRAIL、TXT及二者联合应用对A549细胞的形态学影响和细胞增殖的抑制作用; Annexin ⅤFITC/PI染色,流式细胞术检测A549细胞的凋亡率;半定量RTPCR检测TXT对A549细胞死亡受体DR4、DR5基因表达的影响。结果:①TXT对A549细胞的生长有明显的抑制作用,且呈显著的剂量依赖性,IC50为62.1?ng/ml;而A549对TRAIL不敏感,100?ng/ml的抑制率仅为(6.84±1.14)%,1600?ng/ml的抑制率为(26.10±4.02)%。4?ng/ml TXT与100?ng/ml TRAIL联合应用有显著的协同抑制作用(P<0.05),抑制率为(19.98±4.15)%;②100?ng/ml TRAIL、4?ng/ml TXT单独作用A549细胞24?h的凋亡率为4.44%和12.26%。100?ng/ml TRAIL与4?ng/ml TXT联合作用A549细胞24?h的凋亡率为16.84%;③4?ng/ml TXT作用前后,死亡受体DR4、DR5mRNA表达水平无明显改变。结论:人肺腺癌细胞A549对TRAIL不敏感,但TRAIL与TXT联合有显著的协同诱导细胞凋亡作用,这种协同作用机制与DR4、DR5的表达无关。

Abstract: microscope and by MTT assay, respectively. Annexin ⅤFITC and Propidium iodide(PI) were used to stain A549 cells, and flow cytometry(FCM) was used to determine the apoptosis after the treatment of TRAIL or/and TXT. As well as reverse transcriptionpolymerase chain reaction (RTPCR) was applied to semiquantitatively assay the mRNA expression of death receptors DR4 and DR5 in A549 cells before and after the treatment of subtoxic doses of TXT for different hours. Results: ① The growth inhibition by TXT worked in a dosedependent fashion on A549 cells. The value of IC50 of TXT was 62.1ng/ml. The growth inhibition rate (GIR) was (6.84±1.14)%, and (26.10±4.00)% respectively by TRAIL at the concentration of 100?ng/ml and 1?600?ng/ml and in an acting time of 24 hours, and it was (19.98±4.15)% by TRAIL at 100?ng/ml in the presence of TXT 4?ng/ml. There was significant difference in the growth inhibition rate (GIR) by TRAIL before and after the combination of TXT(P<0.05); ② The apoptotsis rate detected by FCM was 4.44%, and 12.26% respectively after the 24 hours treatment by TRAIL(100?ng/ml) or TXT(4?ng/ml), whereas it was increased to 16.84% after the 24 hours treatment of TRAIL(100?ng/ml) combined with TXT(4?ng/ml); ③ The expression level of DR4 and DR5 mRNA in A549 cells had no significant change after the treatment by TXT(4?ng/ml) for 4 hours and 8 hours (P>0.05). Conclusions: Human lung adenocarcinoma A549 cells are not sensitive to TRAIL, but are sensitive to docetaxel on TRAILinduced apoptosis. The effect of TXT reversal on TRAIL apoptosis pathway is not supposed to be related to the upregulation of death receptors DR4 and DR5 mRNA in A549 cells.

Key words: Lung neoplasms, Tumor necrosis factor, Apoptosis inducing ligand, TXT, Apoptosis

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