【作者】 张萍； 高献书； 周志国； 卢付河； 乔学英； 宋永辉； 杨香然；

【Author】 Zhang Ping GaoXian-Shu Zhou Zhi-Guo Lu Fu-He Qiao Xue-Ying Song Yon-Hui Yang Xiang-Ran Department of Radiation Oncology, the fourth hospital of Hebei medical university & Hebei provincial tumor hospital, Shijiazhuang, 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院放疗一科； 北京大学附属第一医院放疗科；

【摘要】 目的:探讨重组腺病毒-p53抗癌注射液(今又生)、重组人源化表皮生长因子受体单克隆抗体(hR3)单独及联合应用对不同放射敏感性食管癌细胞的作用。方法:用MTT法测定hR3、重组腺病毒-p53抗癌注射液(今又生)及两药在间隔0h、6h、12h、24h各给药组对TE13及放射抗拒性细胞 TE13R120的生长抑制情况及对细胞生长的影响;采用流式细胞术检测两种药物联合作用时对TE13、TE13R120细胞周期分布的影响及对Bax、Bcl-2蛋白表达的影响。结果:hR3、今又生及间隔不同给药时间组均对两种细胞增殖产生抑制作用,与阴性对照组比较有显著性差异(P<0．05)。同期给药组(hR3+今又生)对TE13和TE13R120细胞的抑制作用较明显,抑制率分别为77．93±3．81％、86．09±4．35％,高于 hR3单药组(67．11±4．01％、76．00±6．40％),两者相比无显著性差异(P>0．05),也高于今又生单药组(73．90±3．82％、 84．57±3．83％)两者相比无显著性差异(P>0．05)。两药对放射抗拒性细胞TE13R120的增殖抑制作用高于其亲代TE13, 但未见显著性差异。流式细胞仪分析结果表明,hR3与今又生同时作用后12h,对TE13细胞产生明显的G1期阻滞(G1 期细胞百分比为70．68±3．61％),与对照组相比(G1期细胞百分比为57．89±11．56％)有显著性差异(P<0．05)而S期细胞显著减少,与对照组相比有显著性差异(P<0．05)。而对TE13R120可产生轻度的G1期阻滞,与对照组相比无显著性差异(P>0．05)。TE13及TE13R120在正常生长情况下凋亡率分别为5．20±0．92％、4．60±1．40％,在给予4Gy剂量照射后12h,两种细胞的凋亡率有所增高,分别为5．30±0． 55％、5．40±0．10％,hR3与今又生同时作用后12h,凋亡率为5．63±0．57％、4．93±0．25％,与对照组及照射组相比均无显著性差异(P>0．05)。TE13及TE13R120在正常生长情况下Bax表达相似,给予hR3和今又生共同作用12h后,Bax 表达有所增高,无显著性差异(P>0．05);Bcl-2在两种细胞正常生长时都有较高表达,给予hR3和今又生共同作用12h 后,其表达减少,但无显著性差异(P>0．05)。结论:hR3 与今又生联合应用对不同放射敏感性的食管癌细胞产生了增殖抑制作用,这种增殖抑制作用的发生可能与G1期阻滞有关,其具体机制有待于进一步研究。更多还原

【Abstract】 Objective: The aim of the study was to observe the effect of recombinant adenoviruses-p53 antican-cer injection (Gendicine) combined with a human monoclonal antibody to EGFR (hR3) on esophageal cancer cells with different radiosensitivities. Methods: The inhibition effect of cell growth and the influence by hR3 and Gendicine interval 0. 6. 12. 24h were measured with MTT method. Changes of cell cycle and expressions of Bax and Bcl-2 after treatment with hR3 and Gendicine were detected by FCM. Results: hR3, Gendicine and every experimental group all showed the antiproliferative effect in vitro growth profile of each esophageal cell line and they were significant (P <0.05). Inhibition rate of the simultaneous administration was higher than the hR3 or the Gendicine group. The antiproliferative effect of the simultaneous administration on TE13R120 cells was higher than on TE13 cells, but there was no significant difference. At 12 hours after exposure to hR3 and Gendicine, FCM analysis demonstrated that hR3 and Gendicine treatment induced accu- mutation of TE13 cells in G1, with statistical significance of P <0.05. While there was a slight accumulation of TE13R120 cells in G1 phase and there was no significance (P > 0.05 ). At 12 hours after treatment with hR3 and Gendicine, the pro-apoptotic Bax expression was increased and anti-apoptotic Bcl-2 expression was decreased, with no significant difference of P > 0.05. Conclusion: We observed the anti-proliferation effect of hR3 and Gendicine on different radiosensitivities esophageal cancer cells, which may be concerned with accumulation of cells in G1 phase. But the mechanism needs to be studied further.更多还原