Wang Zhaoxia, Lu Binbin, Wang Teng, De Wei, Shu Yongqian
Cancer Center, The First Affiliated Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, P.R.China.
Zhongguo Fei Ai Za Zhi. 2006 Apr 20;9(2):127-31. doi: 10.3779/j.issn.1009-3419.2006.02.06.
p53 gene is the most commonly mutated gene in lung cancer. p53 mutation results in insensitivity of cells when exposed to chemotherapy. It has been reported that adenovirus-mediated wild-type p53 gene transfection into lung cancer cells can enhance the cytotoxic effect of anti-cancer drugs. The aim of this study is to evaluate the effects of domestic recombinant adenovirus-p53 (Ad-p53, Gendicine) on growth and chemosensitivity of human lung adenocarcinoma cell lines.
Human lung adenocarcinoma cell lines GLC-82 (including mutant p53) and A549 (including wild-type p53) were treated with Ad-p53, cisplatin (DDP) or Ad-p53+DDP respectively. p53 expression was detected by Western blot. The cell growth inhibition was assessed by MTT, and cell cycle and apoptosis were detected by flow cytometry.
High-level p53 expression was detected in Ad-p53 infected GLC-82 and A549 cells by Western blot. There was a dose-dependent and time-dependent inhibition of cell proliferation by Ad-p53. After combined treatment with Ad-p53 (100MOI) and DDP (0.5mg/L) for 72h, the growth inhibition rate of A549 cells was 43.13%±0.72%, which was significantly higher than that in Ad-p53 group ( 23.44%±0.54%, P < 0.001) and DDP group (14.17%±1.39%, P < 0.001); and the growth inhibition rate of GLC-82 cells was 63.73%±0.92%, which was significantly higher than that in Ad-p53 group ( 41.51%±0.59%, P < 0.001) and DDP group (56.11%±1.12%, P < 0.001). Combined administration of Ad-p53 and DDP remarkably arrested A549 and GLC-82 cells in G0-G1, and cells in S phase significantly decreased. Meanwhile the apoptotic rate of A549 cells was 28.99%±1.07% in Ad-p53+DDP group, which was significantly higher than that in Ad-p53 group (15.35%±1.31%, P < 0.001) and DDP group (1.74%±0.77%, P < 0.001). The apoptotic rate of GLC-82 cells was 62.98%±2.43% in Ad-p53+DDP group, which was significantly higher than that in Ad-p53 group (20.88%±0.71%, P < 0.001) and DDP group (6.91%±1.52%, P < 0.001).
Ad-p53 (Gendicine) can inhibit the growth of human lung adenocarcinoma cell lines irrespective of the status of endogenous p53 gene. Its combination with DDP may significantly enhance the chemosensitivity of human lung adenocarcinoma cells to DDP.
p53基因是肺癌中最常发生突变的基因。p53突变导致细胞在接受化疗时不敏感。据报道，腺病毒介导的野生型p53基因转染肺癌细胞可增强抗癌药物的细胞毒性作用。本研究旨在评估国产重组腺病毒-p53（Ad-p53，商品名：今又生）对人肺腺癌细胞系生长及化疗敏感性的影响。
人肺腺癌细胞系GLC-82（含突变型p53）和A549（含野生型p53）分别用Ad-p53、顺铂（DDP）或Ad-p53+DDP处理。采用蛋白质免疫印迹法检测p53表达。采用MTT法评估细胞生长抑制情况，采用流式细胞术检测细胞周期及凋亡情况。
蛋白质免疫印迹法检测显示，Ad-p53感染的GLC-82和A549细胞中检测到高水平的p53表达。Ad-p53对细胞增殖的抑制呈剂量和时间依赖性。Ad-p53（100MOI）与DDP（0.5mg/L）联合处理72小时后，A549细胞的生长抑制率为43.13%±0.72%，显著高于Ad-p53组（23.44%±0.54%，P<0.001）和DDP组（14.17%±1.39%，P<0.001）；GLC-82细胞的生长抑制率为63.73%±0.92%，显著高于Ad-p53组（41.51%±0.59%，P<0.001）和DDP组（56.11%±1.12%，P<0.001）。Ad-p53与DDP联合给药显著使A549和GLC-82细胞停滞于G0-G1期，S期细胞显著减少。同时，Ad-p53+DDP组A549细胞的凋亡率为28.99%±1.07%，显著高于Ad-p53组（15.35%±1.31%，P<0.001）和DDP组（1.74%±0.77%，P<0.001）。Ad-p53+DDP组GLC-82细胞的凋亡率为62.98%±2.43%，显著高于Ad-p53组（20.88%±0.71%，P<0.001）和DDP组（6.91%±1.52%，P<0.001）。
Ad-p53（今又生）可抑制人肺腺癌细胞系的生长，而与内源性p53基因状态无关。其与DDP联合使用可能显著增强人肺腺癌细胞对DDP的化疗敏感性。
