Synergistic cytotoxic effects of recombinant human adenovirus p53 and radiation at various time points in A549 lung adenocarcinoma cells
Synergistic cytotoxic effects of recombinant human adenovirus p53 and radiation at various time points in A549 lung adenocarcinoma cells
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DOI：
10.3892/ol.2012.747
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发表时间：
2012-09-01
期刊：
影响因子：
2.9
通讯作者：
Zou, Hua-Wei
中科院分区：
文献类型：
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作者：
Ma, Jie-Tao;Han, Cheng-Bo;Zou, Hua-Wei
本研究的目的是评估重组人腺病毒p53（rAd-p53; Gendicine）转染和辐射在转染后不同时间点的效果。分析细胞毒性作用和p53蛋白表达水平。将含有人野生型p53基因的rAd-p53导入人肺腺癌细胞系A549中，并用单剂量的6 MeV 4戈伊β射线照射细胞。根据rAd-p53转染与放射治疗之间的时间间隔（RI），将转染rAd-p53的A549细胞分为5组：转染后即刻（0 h-RT）、3 h后（3 h-RT）、6 h后（6 h-RT）、24 h后（24 h-RT）和48 h后（48 h-RT）放射治疗组。将仅转染rAd-p53（Ad-p53）和转染空腺病毒（Ad）的细胞作为两个对照组。转染72小时后，使用MTT测定法和流式细胞术分析细胞活力和生长，并使用蛋白质印迹分析法分析p53蛋白表达。从0 h-RT到48 h-RT，细胞活力逐渐下降，而凋亡细胞百分比和p53蛋白表达逐渐增加。在6 h-RT、24 h-RT和48 h-RT组中，细胞活力抑制率分别为56.7 ± 5.4、60.8 ± 6.0和68.9 ± 6.6，与Ad-p53相比显著更高（40.8 +/- 4.7）、0 h-RT（45.0 +/- 3.5）和3 h-RI组（47.0 +/- 4.3）。RT 6、24、48 h细胞活力抑制率差异无统计学意义（P>0.05）。在凋亡细胞的百分比中观察到类似的变化。6 h-RT组p53蛋白表达水平（0.856 ± 0.092）高于3 h-RT组（0.643 ± 0.089）（t=2.882; P=0.045），但与24 h-RT组（1.193 ± 0.202）无显著性差异。A549细胞活力抑制率和凋亡细胞百分率与p53蛋白表达呈正相关（P
The aim of this study was to evaluate the effects of recombinant human adenovirus p53 (rAd-p53; Gendicine) transfection and radiation at various time points following transfection. Cytotoxic effects and p53 protein expression levels were analyzed. rAd-p53 containing the human wild-type p53 gene was introduced into the human lung adenocarcinoma cell line A549, and cells were irradiated with a single dose of 6 MeV 4 Gy beta rays. According to the time interval between rAd-p53 transfection and radiotherapy (RI), A549-transfected rAd-p53 cells were divided into 5 groups: radiation administered immediately after transfection (0 h-RT) group, after 3 h group (3 h-RT), after 6 h group (6 h-RT), after 24 h group (24 h-RT) and after 48 h group (48 h-RT). Cells with rAd-p53 transfection alone (Ad-p53) and with empty adenovirus (Ad) were included as the two control groups. Following 72 h of transfection, cell viability and growth were analyzed using MTT assays and flow cytometry, and p53 protein expression was analyzed using western blot analysis. From 0 h-RT to 48 h-RT, cell viability gradually decreased, while percentage of apoptotic cells and p53 protein expression gradually increased. The cell viability suppression rates in the 6 h-RT, 24 h-RT and 48 h-RT groups were 56.7 +/- 5.4, 60.8 +/- 6.0 and 68.9 +/- 6.6, respectively, which were significantly greater compared to that of the Ad-p53 (40.8 +/- 4.7), 0 h-RT (45.0 +/- 3.5) and 3 h-RI groups (47.0 +/- 4.3). No statistically significant differences were observed in the cell viability suppression rates among the 6 h-RT, 24 h-RT and 48 h-RT groups (P>0.05). Similar changes were observed in the percentage of apoptotic cells. The p53 protein expression level in the 6 h-RT group (0.856 +/- 0.092) was higher compared to that in the 3 h-RT group (0.643 +/- 0.089) (t=2.882; P=0.045), but not significantly different from that of the 24 h-RT group (1.193 +/- 0.202). The cell viability suppression rate and percentage of apoptotic cells was positively correlated with p53 protein expression in the A549 cells (P