【摘要】 目的:研究重组TRAIL腺病毒(Ad-TRAIL)对人非小细胞肺癌细胞株YTMLC、A549、H460抑制生长和诱导凋亡的作用,探讨Ad-TRAIL用于非小细胞肺癌基因治疗的可能性。方法:培养肿瘤细胞,分别加入Ad-TRAIL、sTRAIL,并设Ad、PBS作对照。倒置显微镜、吖啶橙染色后荧光显微镜、电镜进行细胞形态结构观察;肿瘤细胞生长抑制实验检测细胞存活率,绘制肿瘤细胞生长抑制曲线;DNA凝胶电泳、PI染色、TUNEL染色和流式细胞术检测肿瘤细胞的凋亡;集落形成实验观察肿瘤细胞集落形成能力。结果:Ad-TRAIL组、sTRAIL组与空腺病毒Ad组、PBS阴性对照组相比均明显抑制非小细胞肺癌细胞株YTMLC、A549、H460的增殖。DNA凝胶电泳出现DNA梯形条带;形态学观察发现Ad-TRAIL组的肿瘤细胞具有细胞凋亡的形态;Ad-TRAIL作用的YTMLC细胞有较高比例(8.55%)的亚二倍体凋亡峰,而Ad和PBS组分别为1.08%和0.47%;Ad-TRAIL组被TUNEL标记的细胞比例为10.6%,高于Ad组的1.13%。Ad-TRAIL组的集落数为28±7,而PBS和Ad对照组分别为257±18,193±12。结论:Ad-TRAIL对非小细胞肺癌细胞具有抑制生长和诱导凋亡的作用,Ad-TRAIL有可能用于非小细胞肺癌的基因治疗。更多还原

【Abstract】 Objective: To evaluate the effects of recombinant adenovirus TRAIL (Ad-TRAIL) on proliferation and apoptosis of non-small cell lung cancer (NSCLC) cell line: YTMLC, A549, and H460, so as to assess the feasibility of using Ad-TRAIL for gene therapy of NSCLC. Methods: Tumor cells cultured in presence of Ad-TRAIL or sTRAIL were included in the experimental groups and those cultured in presence of Ad or PBS served as control groups. Morphology observation of cells was performed under inverted microscope, fluorescent microscope with acridine orange staining, and electron microscopy. Cell proliferation inhibiting curves were drawn through cell proliferation inhibiting assay to determine cell survival rate. Tumor cell apoptosis was detected by DNA gel electrophoresis. After stained with PI or TUNEL, the cells were subjected to flow cytometry examination of apoptosis. Clone-forming ability of cells was assessed by agar clonogenic assay. Results: The growth of NSCLC cell lines YTMLC, A549, and H460 cells in Ad-TRAIL and sTRAIL groups was significantly inhibited compared with that in Ad and PBS groups. DNA ladder was revealed on DNA gel electrophoresis and typical apoptosis morphology was observed in Ad-TRAIL group. YTMLC cells in Ad-TRAIL had a higher sub-G1 fraction apoptosis (8.55%) than those in Ad and PBS groups (1.08% and 0.47%, respectively). Cells TUNEL-labeled cells in Ad-TRAIL group (10.6%) were more than those in the Ad group (1.13%). Ad-TRAIL treated YTMLC cells produced (28±7) colonies in the soft agar colonogenic assay; PBS and Ad treated YTMLC cells produced (257±18) and (193±12) colonies, respectively. Conclusion: Ad-TRAIL can inhibit proliferation and induce apoptosis of NSCLC cells in vitro; it might be used for gene therapy of NSCLC.更多还原