GSTθ高表达抑制宫颈癌HeLa细胞的生长和增加放射敏感性

Eehanced Expression of GSTθ Inhibits Growth and Increases Radiosensitivityin Human Uterine Cervix Cancer HeLa Cells

目的了解增加GSTθ表达水平对人类宫颈癌HeLa细胞生长,细胞周期和放疗敏感性的影响。方法通过脂质体LipofectAMINE将pcDNA3-GSTθ真核质粒载体转染获得高GSTθ表达水平的HeLa稳定转染细胞;采用RT-PCR和Western蛋白免疫印迹法检测分别检测GSTθ mRNA和蛋白的表达水平;采用细胞记数和MTT法测定细胞的生长;采用流式细胞仪分析细胞周期的变化。结果GSTθ高水平表达显著地延缓和降低HeLa细胞的生长,并导致G2/M期的阻滞。GSTθ水平提高也增加了细胞对培养基中血清和生长因子的依赖性。另外,GSTθ高水平的表达增加了HeLa细胞对电离辐射的敏感性。结论我们首次证实在HeLa细胞,GSTθ高水平表达可以引起细胞周期阻滞,延缓和降低细胞生长和增加细胞的放射敏感性。这些实验结果将为GSTθ作为一新的靶基因在宫颈癌治疗中的作用提供了实验基础和依据。

Objective To study the effects of glutathione S-transferase theta(GSTθ)on the growth,cell cycle progression and radiaosensitivity of uterine cervix cancer cell line HeLa.Methods Increased expression of GSTθin HeLa cells was performed by stable transfection with a mammalian pcDNA3-GSTθexpression vector.GSTθmRNA and protein were detected by RT-PCR and Western blot assays,respectively.MTT and cell count assays were applied for evaluation of cell growth;and colony formation assay was employed to determine cell survival following -irradiation.Flow cytometry was used to analyze cell cycle arrest.Results Increased expression of GSTθsignificantly caused a G2/M arrest and reduced cell growth.The growth of cells containing GSTθtransfection become to be more dependent on the serum concentration supplied in the growth medium.Finally,GSTθincreased susceptibility of HeLa cells to -irradiation.Conclusion The present results,for the first time,demonstrated that GSTθplays an important role in growth,cell cycle progression and radiosensitivity in human uterine cervix cancer HeLa cells,which will provide evidence for GSTθas a new target in therapy of uterine cervix cancer.