张燕华①, 谢忆山②, 刘少平, 刘诗权, 彭春伟, 邵丽华, 李 雁. 重组人p53腺病毒联合表阿霉素抑制胃癌细胞的体外研究*[J]. 中国肿瘤临床, 2010, 37(18): 1024-1027. DOI: 10.3969/j.issn.1000-8179.2010.18.002
引用本文: 张燕华①, 谢忆山②, 刘少平, 刘诗权, 彭春伟, 邵丽华, 李 雁. 重组人p53腺病毒联合表阿霉素抑制胃癌细胞的体外研究*[J]. 中国肿瘤临床, 2010, 37(18): 1024-1027. DOI: 10.3969/j.issn.1000-8179.2010.18.002
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ZHANG Yanhua1, XIE Yishan2, LIU Shaoping3, LIU Shiquan3, PENG Chunwei3, SHAO Lihua3, LI Yan3. In Vitro Effect of Recombinant Human Adenovirus p53 Combined with Epirubicin on Human Gastric Cancer Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(18): 1024-1027. DOI: 10.3969/j.issn.1000-8179.2010.18.002
Citation: ZHANG Yanhua1, XIE Yishan2, LIU Shaoping3, LIU Shiquan3, PENG Chunwei3, SHAO Lihua3, LI Yan3. In Vitro Effect of Recombinant Human Adenovirus p53 Combined with Epirubicin on Human Gastric Cancer Cells[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2010, 37(18): 1024-1027. DOI: 10.3969/j.issn.1000-8179.2010.18.002
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重组人p53腺病毒联合表阿霉素抑制胃癌细胞的体外研究*

In Vitro Effect of Recombinant Human Adenovirus p53 Combined with Epirubicin on Human Gastric Cancer Cells

    摘要
  • 摘要: 目的：研究基因治疗药重组人p53腺病毒（rAd-p53）与化疗药表阿霉素（EPI）对胃癌细胞的体外作用，探讨基因治疗联合化疗对胃癌的疗效。方法：胃腺癌MGC-803 细胞分对照组、rAd-p 53组、EPI 组、联合用药四组，通过观察细胞生长曲线、流式细胞分析、瑞氏染色、细胞免疫化学染色，研究比较各组细胞生长、细胞周期、细胞形态学、P53蛋白表达。结果：与对照组相比，rAd-p 53在用药1、2、3、6d 后对MGC-803 细胞抑制率为34.52% 、10.53% 、58.98% 、79.17% ，EPI 为60.71% 、67.29% 、73.21% 、95.14% ，联合两药抑制率依次为59.52% 、66.17% 、67.15% 、99.31% ，联合用药抑制率显著高于单药处理（P 均<0.01）。 流式细胞分析发现，rAd-p 53主要将细胞周期阻滞于G2/M期（44.24%），Sub-G 1 凋亡峰升高，联合EPI 治疗G2/M期（81.73%）及Sub-G 1 峰增高更明显（P 均<0.01）。 瑞氏染色可见，rAd-p 53治疗组MGC-803 细胞仅有少数聚集，细胞形态不规则，胞质深染，可见凋亡细胞；EPI组细胞呈小片状聚集，形态结构较完整，与对照组无明显差别；联合用药组细胞数明显减少，形态不规则，胞质深染，凋亡细胞数更多。免疫细胞化学染色可见，P53蛋白于rAd-p 53治疗组细胞核及胞质中表达，联合EPI 作用效果更显著；对照组与EPI 组P53蛋白于细胞核表达。结论：rAd-p 53对胃癌MGC-803 细胞有抑制作用，促进细胞凋亡，可提高胃癌MGC-803 细胞对化疗药EPI 的敏感性，二者有协同作用。

     

    Abstract: Objective:To study the effect of recombinant human adenovirus p 53injection (rAd-p 53) and Epirubicin Hy -drochloride (EPI) on human gastric cancer cells in vitro, and explore the therapeutic effect of gene therapy combined with chemotherapy on gastric cancer. Methods:The MGC- 803 human gastric adenocarcinoma cell line was treated with rAd-p 53or/and EPI or left untreated as a blank control. Cell growth, flow cytometry, Wright's staining, and cyto-immuno-chemistry were used to analyze the therapeutic efficacy of the different treatment groups. Results: After treating MGC- 803 on days 1, 2, 3 & 6, the cell inhibition rates were 34.52%, 10.53%, 58.98% and 79.17%, respectively, for rAd-p53group; 60.71%,67.29%,73.21% and 95.14%, respectively, for EPI group; and59.52%,66.17%,67.15% and 99.31%, respective-ly, for the combined group. The combined group was significantly higher than that of single agent groups (P<0.01). Based on the flow cytometry, we found that rAd-p53blocked the cell cycle at G 2/M phase ( 44.243 %) and a higher Sub-G1 peak was observed. In the combined group, the G2/M phase ( 81.734 % ) and Sub-G1 peak were significantly evident (P< 0.01). Decreased cell number, abnormal cell morphology and apoptotic cells were found under Wright's staining, and p53protein stained positively in the nucleus and cytoplasm by immunochemical analysis in rAd-p53treated group, especially in the combined treatment group. The EPI group was not different from the blank control. Conclusion:The gene therapy agent rAd-p 53has an inhibiting and apoptotic effect on MGC- 803 cells, thus it could enhance the chemotherapeutic effect of EPI on MGC-803 cells, suggesting synergism between gene therapy and chemotherapy.

     

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