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背景与目的在肺癌中,p53突变是最常见的基因改变之一,p53基因的突变常导致细胞对化疗不敏感。有研究表明导入野生型p53基因能增加化疗药物的敏感性。本研究的目的是探讨重组人p53腺病毒注射液(Adp53,今又生,Gendicine)对人肺腺癌细胞生长及化疗敏感性的影响。方法将重组腺病毒载体所携带的野生型p53基因导入人肺腺癌细胞株GLC82(含突变型p53基因)及A549(含野生型p53基因),并联合应用化疗药物顺铂(DDP),通过Westernblot法分析外源野生型p53基因在细胞内的表达,MTT法和流式细胞术观察对细胞生长及细胞周期、凋亡的影响。结果通过Westernblot证实了外源p53基因能在GLC82及A549细胞中高效表达。MTT法观察到Adp53对肺癌细胞的抑制作用呈时间依赖性和剂量依赖性效应。100MOIAdp53与0.5mg/LDDP联合应用后72h,对A549细胞的生长抑制率达43.13%±0.72%,显著高于单用Adp53组(23.44%±0.54%,P<0.001)和DDP组(14.17%±1.39%,P<0.001);对GLC82细胞生长的抑制率达63.73%±0.92%,显著高于单用Adp53组(41.51%±0.59%,P<0.001)和DDP组(56.11%±1.12%,P<0.001)。流式细胞术检测结果显示,Adp53与DDP联合应用能使细胞阻滞于G0G1期,S期细胞比例明显减少。Adp53+DDP组A549细胞凋亡率为28.99%±1.07%,显著高于单用Adp53组(15.35%±1.31%,P<0.001)和DDP组(1.74%±0.77%,P<0.001);Adp53+DDP组GLC82细胞凋亡率为62.98%±2.43%,显著高于单用Adp53组(20.88%±0.71%,P<0.001)和DDP组(6.91%±1.52%,P<0.001)。结论重组人p53腺病毒注射液能抑制肺腺癌细胞的生长,并不受内源性p53状态的影响。它与抗癌药DDP联用能显著增加肺腺癌细胞的化疗敏感性。
Background and Aim p53 mutations are one of the most common genetic alterations in lung cancer. Mutations in the p53 gene often result in cells being less sensitive to chemotherapy. Studies have shown that the introduction of wild-type p53 gene can increase the sensitivity of chemotherapeutic drugs. The purpose of this study was to investigate the effect of recombinant adenovirus p53 (Adp53, Gendicine) on human lung adenocarcinoma cell growth and chemosensitivity. Methods The wild-type p53 gene in recombinant adenovirus vector was transfected into human lung adenocarcinoma cell line GLC82 (including mutant p53 gene) and A549 (including wild-type p53 gene), and combined with chemotherapeutic drug cisplatin (DDP) Western blot analysis of exogenous wild-type p53 gene expression in cells, MTT assay and flow cytometry observation of cell growth and cell cycle, apoptosis. The results confirmed by Westernblot exogenous p53 gene can be highly expressed in GLC82 and A549 cells. MTT assay showed that the inhibitory effect of Adp53 on lung cancer cells in a time-dependent and dose-dependent manner. The growth inhibition rate of A549 cells was 43.13% ± 0.72% at 72h after combination of 100MOIAdp53 and 0.5mg / LDDP, significantly higher than that of AdP53 alone group (23.44% ± 0.54%, P <0.001) and DDP group (14.17% ± 1.39%, P <0.001). The inhibition rate of GLC82 cells was 63.73% ± 0.92%, which was significantly higher than that of Adp53 group (41.51% ± 0.59%, P <0.001) P <0.001). Flow cytometry results showed that, Adp53 combined with DDP can make cell arrest in G0G1 phase, S phase cells decreased significantly. The apoptosis rate of A549 cells in Adp53 + DDP group was 28.99% ± 1.07%, significantly higher than that in Adp53 group (15.35% ± 1.31%, P <0.001) and DDP group (1.74% ± 0.77%, P <0.001) The apoptosis rate of GLC82 + DDP group was 62.98% ± 2.43%, which was significantly higher than that of Adp53 group (20.88% ± 0.71%, P <0.001) and DDP group (6.91% ± 1.52%, P <0.001). Conclusion Recombinant human p53 adenovirus injection can inhibit the growth of lung adenocarcinoma cells and is not affected by the endogenous p53 status. It combined with anti-cancer drug DDP can significantly increase the chemosensitivity of lung adenocarcinoma cells.