尼妥珠单抗联合X射线照射对肺腺癌细胞A549的作用

颜莹; 刘园; 高纯子; 李瑛; 韩波

doi:10.3969/j.issn.1000-8179.2011.08.004

摘要: 目的：探讨表皮生长因子（Epidermal growth factor receptor， EGFR）受体抑制剂尼妥珠单抗（h-R3）联合X射线照射，对肺腺癌细胞系的A549细胞的作用。方法：肺腺癌细胞系A549细胞常规培养48 h后，分为对照组，单纯h-R3组，单纯照射组，h-R3联合照射组。h-R3组分别以终浓度0.5、 5、 50、 100、 1 000 μg/mL处理后继续培养24 h，照射组采用Varian-6MV直线加速器X射线照射2、 4、 8、 10 Gy后继续培养24 h； h-R3联合照射组在照射前以50 μg/mL浓度h-R3处理后继续培养24 h。收集各组细胞进行甲基噻唑基四唑（MTT）法检测。测定细胞的光密度值（Optical Density， OD），绘制细胞存活情况，并在倒置显微镜下观察细胞形态变化。结果：对照组A549细胞的OD值为0.786 7±0.076 4，不同浓度h-R3组的OD值分别为0.793 6±0.084 6， 0.823 4±0.068 8， 0.760 3±0.057 3， 0.790 2±0.063 7， 0.820 8±0.077 0，与对照组比较无明显差异（P>0.05）；不同剂量照射组OD值分别为0.246 0±0.023 1， 0.330 8±0.033 6， 0.343 0±0.028 1， 0.317 5±0.030 7，明显低于对照组（P<0.05）； 50 μg/mL浓度h-R3联合不同剂量照射组OD值分别为0.192 3±0.014 6， 0.231 8±0.020 6， 0.293 0±0.024 3， 0.193 7±0.021 8，与单纯照射组相比，其OD值均明显减少（P<0.01）； h-R3联合照射组的A549细胞形态变化明显，存活细胞数明显少于单纯照射组。结论：尼妥珠单抗（h-R3）单独应用对A549细胞的生长增殖未见明显抑制作用，但增强了照射对A549细胞的生长抑制作用，其放射增敏的作用机制值得进行分子生物学水平的深入研究。

Abstract: The Impact of Nimotuzumab (hR-3) Combined with Radiation on Human PulmonaryAdenocarcinoma A549 CellsYing YAN, Yuan LIU, Chunzi GAO, Ying LI, Bo HANCorrespondence to: Bo HAN, E-mail: baohan1964@163.comDepartment of Oncology, The First Affiliated Hospital of Harbin Medical University, Harbin 150001, ChinaThis study was supported by Beijing Science and Technology Development Program (No.D00802000290803)Abstract Objective: To investigate the effects of Nimotuzumab ( hR-3 ) combined with irradiation on pulmonary adenocarcino-ma A549 cells. Methods: Human pulmonary adenocarcinoma cells A549 was cultured in vitro for 48 h and was divided into 4 groups:control group, the irradiation group, h-R3 group, and h-R3 combined with radiation group. The h-R3 group were treated with h-R3 atdifferent concentrations ( the final concentrations were 0.5, 5, 50, 100, and 1000 μg/mL) and then were cultured for 24 h. The irradia-tion group received irradiation of 2, 4, 8, and 10 Gy X-ray by 6MV-X ray and then were cultured for 24 h. The combined group re-ceived h-R3 at 50 μg/mL and were cultured for 24 h, followed by radiation at doses of 2, 4, 8, and 10 Gy and continuous culture of 24h. MTT assay was used to evaluate the amount of survival cells. The characteristics of morphological changes were observed by invert-ed microscope. Results: The OD value of A549 cells in the control group was 0.7867 ± 0.0764. In the h-R3 group, the OD values ofA549 cells treated with h-R3 at 0.5, 5, 50, 100, and 1000 μg/mL were 0.7936 ± 0.0846, 0.8234 ± 0.0688, 0.7603 ± 0.0573, 0.7902 ±0.0637, and 0.8208 ± 0.077, respectively. No significant difference was found in OD values between the control group and the h-R3group ( P < 0.05 ). In the irradiation group, the OD values of A549 cells treated with irradiation of 2, 4, 8, and 10 Gy were 0.2460 ±0.0231, 0.3308 ± 0.0336, 0.3430 ± 0.0281, and 0.3175 ± 0.0307, respectively, significantly lower than in the control group ( P < 0.05 ).In the combined group, the OD values were 0.1923±0.0146, 0.2318±0.0206, 0.2930±0.0243, and 0.1937±0.0218, respectively, signifi-cantly lower than those in the irradiation group ( P < 0.01 ). In the combined group, obvious morphological changes were observed inA549 cells, and there were fewer viable cells than in the radiation group, with a statistical significance ( P < 0.05 ). Conclusion: Nimo-tuzumab ( hR-3 ) can increase the inhibitory effect of irradiation on human pulmonary adenocarcinoma A549 cells. Further study is war-ranted to elucidate the corresponding molecular mechanisms.Keywords Nimotuzumab (h-R3); Lung adenocarcinoma A549 cells; X ray; Radiotherapy; MTT