中华放射医学与防护杂志

张洪波,张红雁,王宏梅,等.RNA干扰肝癌MHCC97-H细胞N-Ras基因对放射敏感性的影响[J].中华放射医学与防护杂志,2011,31(3):252-255.ZHANG Hong-bo,ZHANG Hong-yan,WANG Hong-mei,et al.Effect of N-Ras by RNA interference on radiosensitivity of hepatoma carcinoma cell MHCC97-H line[J].Chin J Radiol Med Prot,2011,31(3):252-255

RNA干扰肝癌MHCC97-H细胞N-Ras基因对放射敏感性的影响

Effect of N-Ras by RNA interference on radiosensitivity of hepatoma carcinoma cell MHCC97-H line

投稿时间：2010-10-27

DOI：10.3760/cma.j.issn.0254-5098.2011.03.002

中文关键词: 放射敏感性 N-Ras基因 RNA干扰肝肿瘤

英文关键词:Radiosensitivity N-Ras gene RNA interference Hepatoma carcinoma

基金项目:国家自然科学基金(30772530);安徽省卫生厅医学科研资助项目(09B109)

作者	单位
张洪波	230000 合肥,安徽省立医院肿瘤放疗科
张红雁	230000 合肥,安徽省立医院肿瘤放疗科
王宏梅	南方医科大学南方医院肿瘤放疗科
罗文广	230000 合肥,安徽省立医院肿瘤放疗科
陈龙华	南方医科大学南方医院肿瘤放疗科

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中文摘要:

目的探讨RNA干扰(RNAi)N-Ras基因增强人肝癌MHCC-97细胞的放射敏感性。方法通过构建干扰N-Ras基因siRNA,采用免疫组织化学法、实时荧光定量RT-PCR、Western blot、MTT法观察RNAi前后肝癌MHCC97-H细胞RNA水平、蛋白水平、生长情况等变化,照射后用细胞克隆计数实验检测放射敏感性变化。结果 MHCC97-H细胞株RNAi后mRNA表达抑制率为96.9%±0.159%,RNAi前后差异有统计学意义(t=40.377, P <0.05),蛋白表达抑制率为89.8%±0.012%,RNAi前后差异有统计学意义(t=31.595, P <0.05),免疫组织化学显示90%表达被抑制,细胞生长抑制率为21.9%,RNAi前后差异均有统计学意义(F =4.63, P <0.05)。MHCC97-H细胞RNAi后增敏比(SER)=1.15。结论 RNAi肝癌MHCC97-H细胞N-Ras基因可以达到放射增敏的目的。

英文摘要:

Objective To investigate the radiosensitivity of silencing N-Ras by RNA interference in hepatoma carcinoma cell MHCC-97. Methods N-Ras RNA interference(RNAi) vector was constructed by using pcDNA 6.2-GW/EmGFP-mir plamid. The RNAi effect was detected by RT-PCR,Western bolt,immunohistochemisty and MTT method.Survival curve for each cell line were obtained by measuring the clone forming abilities of irradiated cell populations. Results After silencing the N-Ras by RNAi,The expression level of N-Ras mRNA,N-Ras protein, immunohistochemisty were decreased 96.9%±0.159%(t=40.377, P <0.05),89.8%±0.012%(t=31.595, P <0.05), 90%, respectively.and The survival of hepatoma carcinoma cell MHCC-97 line were inhibited 21.9%(F =4.63, P <0.05).Which have significant difference in statistics.The SER of hepatoma carcinoma cell MHCC-97 line after interference was 1.15. Conclusions RNAi targeting silence N-Ras may increase the radiosensitivity of hepatoma carcinoma cell MHCC-97 line.

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