人凋亡相关新基因PNAS-4与放射联合治疗Lewis肺癌的实验研究

Radiosensitivity of Lung Neoplasms by Liposome-mediated hPNAS-4 Gene,A Novel Apoptosis-related Human Gene

目的:探讨人凋亡相关新基因PNAS-4(hPNAS-4)基因通过脂质体转染至Lewis肺癌LL2细胞后对放射治疗的增敏作用。方法:用脂质体介导的转染技术,将hPNAS-4基因导入Lewis肺癌LL2细胞中,Western blot鉴定其过表达后,观察X射线照射对其集落形成的影响;流式细胞仪检测hPNAS-4基因或/和放疗(0,1,2,4,6 Gy)对LL2细胞生长抑制及凋亡的影响。结果:通过Western blot证实了hPNAS-4基因在LL2细胞中的过表达。Lip-hPNAS-4加放射治疗处理组细胞的生存能力明显降低,肿瘤细胞的克隆形成明显减少,流式细胞术检测体外培养的肿瘤细胞凋亡明显增加。结论:hPNAS-4基因联合放射治疗能产生协同抗肿瘤效应。

Objective： To explore the efficacy of a novel apoptosis-related human gene （hPNAS-4 gene） therapy combined with radiotherapy against LL2 Lewis lung cancer cells, and whether Lip-hPNAS-4 could enhance the antitumot sensitivity to radiation in vitro. Methods： LL2 ceils were exposed to radiation followed by liposome-mediated transfection, hPNAS-4 gene was introduced into LL2 cells by lipofectin-mediated gene transfeetion. The expression of hPNAS-4 gene was detected by Western blot. The colony formation of transfected cells was observed after irradiation with X-rays; and flow cytometric （fluorescence-activated cell sorting） analysis examined the influence of the hPNAS-4 gene or/and the radiotherapy （0, 1, 2, 4, 6 Gy） to LL2 cell. Results： The overexpression of hPNAS-4 gene into LL2 cells was confirmed by Western blot. Lip-hPNAS-4 plus radiation therapy treatment led to more obvious cell disease, significantly decreased the viability of cells and reduced the formation of tumor cell cloning, and flow cytometric detection of tumor cells in vitro indicated significantly increased apoptosis. Conclusion： hPNAS-4 gene can enhance the sen- sitivity of LL2 Lewis lung cancer cells to radiotherapy. The combination of hPNAS-4 gene therapy and radiotherapy may be more effective in lung cancer treatment.