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目的分析重组人p53腺病毒(即rAd-p53)与表阿霉素(EPI)对胃癌细胞的抑制作用。方法将胃癌细胞(MGC-803)平均分为对照组、EPI组、rAd-p53组、两药联合组,通过观察胃癌细胞的增殖曲线、细胞内瑞氏染色和化学染色,对各组的细胞繁殖周期及细胞形态、p53基因的表达进行比较。结果与其对照组相比,两药联合组的抑制百分率为:59.93%、67.05%、68.65%、99.40%,两药联合运用的抑制率与单药处理的抑制率相比差异具有统计学意义(P<0.01)。流式细胞的分析发现,重组人p53腺病毒主要使胃癌细胞停留于G2/M期,联合EPI对细胞G2/M期的阻滞作用,使Sub-G1峰极高。而通过瑞氏染色可得,EPI组的细胞呈现聚集状,结构和形态较为完整;rAd-p53组则只有少量聚集,并且细胞形态破坏,细胞凋亡。两药联合组的细胞数急剧减少,形态结构破坏,凋亡数目比rAd-p53组明显增多。对细胞进行免疫化学染色可得,rAd-p53组与两药联合组的细胞核以及细胞质之中均有p53蛋白的表达,两药联用组表达更明显;而对照组与EPI组中,p53蛋白只于细胞核中表达。结论重组人p53腺病毒联合表阿霉素对胃癌细胞的抑制作用十分显著,在临床应用中值得推广。
Objective To analyze the inhibitory effect of recombinant human p53 adenovirus (rAd-p53) and epirubicin (EPI) on gastric cancer cells. Methods The gastric cancer cells (MGC-803) were equally divided into control group, EPI group, rAd-p53 group and two drug combination groups. By observing the proliferation curve, intracellular Wright’s staining and chemical staining of gastric cancer cells, Reproductive cycle and cell morphology, p53 gene expression were compared. Results Compared with the control group, the inhibition rates of the combination of the two drugs were 59.93%, 67.05%, 68.65% and 99.40%, respectively. The inhibition rates of the combination of the two drugs were statistically significant compared with the single drug treatment ( P <0.01). Flow cytometry analysis showed that the recombinant human p53 adenovirus mainly caused gastric cancer cells to stay in the G2 / M phase. Combined with the blockade of G2 / M phase of EPI, the Sub-G1 peak was extremely high. By Wright’s staining, EPI group of cells showed aggregated, the structure and morphology is more complete; rAd-p53 group only a small amount of aggregation, and cell morphology damage, apoptosis. The combination of the two drugs in the sharp decline in the number of cells, morphological damage, the number of apoptosis than rAd-p53 group increased significantly. Immunohistochemical staining of cells showed that p53 protein was expressed in the nucleus and cytoplasm of both rAd-p53 group and two drug combination groups, and the expression of p53 protein was more obvious in the combination group and the control group. In the control group and the EPI group, p53 protein Only expressed in the nucleus. Conclusion Recombinant human p53 adenovirus combined with epirubicin has a significant inhibitory effect on gastric cancer cells and is worth promoting in clinical application.