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目的:探讨RNA干扰RelB基因对鼠RM-1前列腺癌细胞株放射敏感性的影响及其机制。方法:利用靶向RelB基因的慢病毒载体(pLentilox-sh-RelB),脂质体介导转染RM-1细胞后进行放射(2,4,6,和8Gy剂量)处理培养,分别采用RT-PCR、Western印迹法及流式细胞术等方法检测RelB的mRNA和蛋白表达,锰超氧化物歧化酶(Mn-SOD)活力及细胞放射敏感性的变化。结果:转染后放射处理培养,siRelB-RM-1组RelB的mRNA和蛋白表达水平明显低于siVector-RM-1组和nontrans-RM-1组(P<0.05);放射处理培养后siRelB-RM-1组细胞凋亡百分率明显高于siVector-RM-1组和nontrans-RM-1组(P<0.05);培养后siRelB-RM-1组细胞Mn-SOD活力下降,放射增敏比为5.13。结论:RNA干扰RelB基因可增强鼠RM-1前列腺癌细胞株放射后的放射敏感性,其机制可能与RNA干扰RelB基因,抑制细胞增殖,降低Mn-SOD活力和诱导凋亡有关。
Objective: To investigate the effect of RelB gene RNA interference on radiosensitivity of murine RM-1 prostate cancer cell line and its mechanism. Methods: The lentiviral vector targeting RelB gene (pLentilox-sh-RelB) was transfected into RM-1 cells by lipofectamine. The cells were treated with radiation (2,4,6 and 8 Gy doses) The mRNA and protein expression of RelB, the activity of manganese superoxide dismutase (Mn-SOD) and the radiosensitivity of cells were detected by PCR, Western blot and flow cytometry. Results: After transfection, the mRNA and protein levels of RelB in siRelB-RM-1 group were significantly lower than those in siVector-RM-1 group and nontrans-RM-1 group (P <0.05) The percentage of apoptosis in RM-1 group was significantly higher than that in siVector-RM-1 group and nontrans-RM-1 group (P <0.05). After cultured, the activity of Mn-SOD in siRelB-RM- 5.13. CONCLUSION: RNA interference of RelB gene can enhance the radiosensitivity of RM-1 prostate cancer cells after radiotherapy. The mechanism may be related to RNA interference with RelB gene, inhibition of cell proliferation, decrease of Mn-SOD activity and induction of apoptosis.