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放疗联合pGL3/EGR1-Bax基因促进肺癌H460细胞凋亡及Bax基因表达的研究

Bax Expression and H460 Apoptosis Induced by Recombined Plasmid pGL3/EGR1-Bax Combined with Radiotherapy in Lung Cancer

  • 摘要: 目的 探讨重组质粒pGL3/EGR1-Bax基因联合放射治疗对肺癌H460细胞的凋亡作用及对Bax 基因表达的影响。方法 将pGL3/EGR1-Bax质粒转染肺癌H460细胞,分组进行射线照射。RT-PCR检测各组细胞Bax基因的表达,蛋白免疫印迹检测Bax蛋白的表达;用流式细胞仪检测各组H460细胞凋亡情况。结果 经酶切鉴定pGL3/EGR1-Bax重组质粒构建正确,基因联合放疗能促进Bax基因及蛋白的表达(与对照组比P<0.01,与空载体放疗组比P<0.05);基因放射治疗组H460细胞凋亡率明显高于对照组及空载体放疗组(P<0.01,P<0.05)。结论 射线可通过诱导pGL3/EGR1-Bax重组质粒中Bax 基因在肺癌H460细胞表达增强,从而显著提高H460细胞的凋亡率,为临床肺癌的基因放射治疗奠定实验基础。

     

    Abstract: Abstract:Objective To investigate Bax expression and the apoptosis of H460 cell induced by recombined plasmid pGL3/EGR1-Bax combined with radiotherapy in lung cancer. Methods Lung cancer cell line H460 was transfected with pGL3/EGR1-Bax plasmids and irradiated in groups. Bax expression was detected by RT-PCR and Western blot. The apoptosis of H460 cell was detected by flow cytometry. Results Restriction enzyme digestion showed pGL3/EGR1-Bax was correctly constructed. Gene-radiotherapy could significantly enhance the expression of Bax gene and protein in H460 cells (P < 0.01,compared with the control group;P< 0.05, compared with the empty vector irradiation group). The apoptosis of H460 cell in pGL3/EGR1-Bax gene therapy plus irradiation group was higher than that in either plasmid or irradiation group alone (P<0.05). Conclusion Irradiation could induce Bax expression in pGL3/EGR1-Bax plasmid in H460 cells, to significantly enhance the apoptosis of H460 cells, which may establish important experimental basis for generadiotherapy for lung cancer.

     

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