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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。
Objective: To investigate the effect of recombinant TRAIL adenovirus (Ad-TRAIL) on the growth inhibition and apoptosis induction of human non-small cell lung cancer cell lines YTMLC, A549 and H460, and explore the possibility of using Ad-TRAIL for gene therapy of non-small cell lung cancer. Methods: Tumor cells were cultured and added with Ad-TRAIL and sTRAIL respectively. Ad and PBS were used as controls. The morphological changes of the cells were observed under inverted microscope, acridine orange staining and fluorescence microscopy. The cell growth inhibition was used to detect the cell viability and the tumor growth inhibition curve was drawn. DNA gel electrophoresis, PI staining, TUNEL staining and flow cytometry The apoptosis of tumor cells was detected; colony formation assay was used to observe the ability of tumor cells to colonize. Results: Ad-TRAIL group and sTRAIL group significantly inhibited the proliferation of non-small cell lung cancer cell lines YTMLC, A549 and H460 compared with empty adenovirus Ad and PBS negative control groups. DNA ladder electrophoresis showed a DNA ladder; Morphological observation found that Ad-TRAIL group of tumor cells with apoptotic morphology; Ad-TRAIL YTMLC cells a higher proportion (8.55%) of the sub-diploid apoptosis Peak, while Ad and PBS groups were 1.08% and 0.47%, respectively. The percentage of TUNEL-labeled cells in Ad-TRAIL group was 10.6%, which was higher than that in Ad group (1.13%). The number of colonies in the Ad-TRAIL group was 28 ± 7, while the PBS and Ad control groups were 257 ± 18, 193 ± 12, respectively. Conclusion: Ad-TRAIL can inhibit growth and induce apoptosis in non-small cell lung cancer cells. Ad-TRAIL may be used in gene therapy for non-small cell lung cancer.