中华放射医学与防护杂志

王佩国,刘志艳,魏枫,等.反义表皮生长因子受体对人肺癌细胞放射敏感性的影响[J].中华放射医学与防护杂志,2008,28(4):361-364.WANG Pei-guo,LIU Zhi-yan,WEI feng,et al.Experimental study of antisense epidermal growth factor receptor enhancing the radiosensitivity of human lung cancer cell line spc-a-1[J].Chin J Radiol Med Prot,2008,28(4):361-364

反义表皮生长因子受体对人肺癌细胞放射敏感性的影响

Experimental study of antisense epidermal growth factor receptor enhancing the radiosensitivity of human lung cancer cell line spc-a-1

投稿时间：2007-12-26

DOI：

中文关键词: 肺腺癌反义表皮生长因子受体放射敏感性

英文关键词:Lung adenocarcinoma Antisense EGFR Radiosensitivity

基金项目:

作者	单位	E-mail
王佩国	300060 天津医科大学附属肿瘤医院放疗科
刘志艳	300060 天津医科大学附属肿瘤医院放疗科
魏枫	300060 天津医科大学附属肿瘤医院生物治疗科
于津浦	300060 天津医科大学附属肿瘤医院生物治疗科
史玉荣	300060 天津医科大学附属肿瘤医院中心实验室
王平	300060 天津医科大学附属肿瘤医院放疗科	wangping908@yahoo.com.cn

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中文摘要:

目的观察反义表皮生长因子受体(EGFR)对人肺腺癌细胞系放射敏感性的影响。方法在脂质体介导下用质粒pcDNA3-反义EGFR(pcDNA3-antiEGFR)转染spc-a-1细胞,并以未转染(对照组)和空质粒pcDNA3转染(pcDNA3组)细胞作对照。用G418筛选稳定表达的细胞克隆,以RT-PCR和Western blot检测转染后EGFR的mRNA及蛋白表达抑制情况,流式细胞仪检测转染后细胞周期及单次照射8 Gy后各组细胞凋亡比例。3组细胞分别给予6 MV X射线照射0、2、4、6和8 Gy,计算克隆形成率,拟合生长曲线。结果pcDNA3-antiEGFR组与对照组、pcDNA3组比较,EGFR mRNA和蛋白表达量明显减少,G₂/M期比例分别为(29.53±1.91)％、(13.7±1.30)%和(12.40±1.34)％,单次照射8Gy后,3个组细胞的凋亡率分别为(39.24±1.57)%、(13.79±0.63)%和(15.02±0.85)%。与对照组相比较,pcDNA3-antiEGFR组细胞的D₀、D_q、SF₂值分别由2.11、2.49和0.84降至1.19、0.15和0.32,提示抑制EGFR表达,可降低spc-a-1细胞对射线所致亚致死性损伤的修复能力。结论反义EGFR可以降低人肺癌spc-a-1细胞中EGFR的表达,改变细胞周期分布,促进凋亡,降低亚致死性损伤的修复能力,增加肺癌细胞系spc-a-1的放射敏感性。

英文摘要:

Objective To explore whether antisense-EGFR could enhance the radiosensitivity of human lung cancer spc-a-1 cell line. Methods The spc-a-1 cells were transfected with antisense-EGFR-pcDNA3 by lipofectamine 2000(pcDNA3 antiEGFR group). Two other groups were used for comparison: control group (spc-a-1 cell without transfection) and pcDNA3 group (spc-a-1 cell transfected with pcDNA3 which did not contain antisense EGFR). Cell clones that stable expressing antisense-EGFR was selected with G418 and the suppression of the expression of EGFR mRNA and protein were detected by RT-PCR and Western blot. The influence of antisense-EGFR on cell cycle was testified by flow cytometry assay. The cell apoptosis was analyzed by flow cytometry after 8 Gy irradiation. Further, cells of each group were irradiated with X-rays at the dose of 0, 2, 4, 6 and 8 Gy. Dose-survival curve of each group was established by colony-forming assay. Results The expression of EGFR mRNA and protein were significantly inhibited after antisense-EGFR- pcDNA3 transfection. The cells arrested at the G₂/M phase in the pcDNA3 antiEGFR group, control group and pcDNA3 group were (29.53±1.91)％, (13.7±1.30)% and (12.40±1.34)％,respectively. The apoptosis index of spc-a-1 cells in the antisense-EGFR combined with irradiation group was obviously higher than that of the comparable groups [(39.24±1.57)%, (13.79±0.63)% and (15.02±0.85%)]. The values of D₀, D_q, SF₂ of pcDNA3 antiEGFR group declined obviously compared with the control group(2.11, 2.49, 0.84 vs 1.19, 0.15, 0.32).Conclusions Antisense-EGFR could induce the G₂/M cell cycle arrest, promote cell apoptosis and inhibit the ability of sublethal cell damage repair induced by irradiation, so that it could significantly improve the radiosensitivity of spc-a-1 cell in vitro.

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