国际肿瘤学杂志

国际肿瘤学杂志 ›› 2013, Vol. 40 ›› Issue (4): 298-302.

• 论著 • 上一篇    下一篇

p53凋亡刺激蛋白2对饥饿诱导的大肠癌HCT116 p53+/+ 细胞凋亡、周期和自噬的影响

侯庆生, 丁渭, 陈德喜, 韩悦, 张玉林, 郭洪亮   

  1. 250117济南，山东省肿瘤医院外科四病区（侯庆生、韩悦、郭洪亮）；济南大学 山东省医学科学院医学与生命科学学院（侯庆生、韩悦）；首都医科大学附属北京佑安医院感染科（丁渭、陈德喜、张玉林）
  • 出版日期:2013-04-08 发布日期:2013-03-27
  • 通讯作者: 郭洪亮，E-mail: hliangbb@163.com E-mail:hliangbb@163.com

The role of ASPP2 in the apoptosis, cell cycle and autophagy of starvation-induced HCT116 p53+/+ cell line

HOU  Qing-Sheng, DING  Wei, CHEN  De-Xi, HAN  Yue, ZHANG  Yu-Lin, GUO  Hong-Liang   

  1. Department of General Surgery, Cancer Hospital of Shandong Province, Jinan 250117, China
  • Online:2013-04-08 Published:2013-03-27
  • Contact: GUO Hong-liang E-mail:hliangbb@163.com

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摘要： 目的 探讨p53凋亡刺激蛋白2（ASPP2）对饥饿诱导的大肠癌HCT116 p53+/+（p53野生型）细胞凋亡、周期和自噬的影响。方法 实验分6组：①对照组 ②绿色荧光蛋白腺病毒（rAd-GFP）感染组 ③ ASPP2腺病毒（rAd-ASPP2）感染组 ④饥饿处理组 ⑤GFP+饥饿组 ⑥ ASPP2+饥饿组 。利用rAd-ASPP2感染使细胞过表达ASPP2基因。无血清培养基培养24 h诱导凋亡、自噬和细胞周期改变。Calcein/PI吸收试验观察各组细胞调亡水平。细胞转染红色荧光蛋白标记的CFP-Lc3自噬质粒，荧光显微镜下观察各组细胞自噬水平。流式细胞术观察细胞周期改变。组间比较采用单因素方差分析进行统计学分析。结果  ASPP2过表达显著促进了饥饿诱导的细胞凋亡（各组细胞的凋亡率:GFP+饥饿组10.00%±1.42%，ASPP2+饥饿组18.44%±2.06%， P=0.000）、自噬（各组细胞的自噬发生率:GFP+饥饿组35.00%±5.34%，ASPP2+饥饿组57.61%±6.06%， P=0.000）、及G2-M期阻滞，但无饥饿诱导时ASPP2过表达使G0-G1、G2-M期都发生阻滞。结论 ASPP2过表达促进饥饿24 h诱导的大肠癌HCT116 p53+/+细胞凋亡和自噬，显著改变细胞周期进程。

关键词: 细胞凋亡, 细胞周期, 自噬, p53凋亡刺激蛋白2

Abstract: Objective To investigate the role of apoptosis stimulating protein 2 of p53（ASPP2）in the apoptosis, cell cycle and autophagy of starvation-induced colorectal cancer HCT116 p53+/+ (p53 wild-type) cell line. Methods Six groups were included: ⑴ control group; ⑵ green fluorescent protein adenovirus (rAd-GFP) infection group; ⑶ ASPP2 adenovirus (rAd-ASPP2) infection group; ⑷ starvation group; ⑸GFP+ starvation group; ⑹ASPP2+ starvation group. HCT116 cells were infected with ASPP2 adenovirus (rAd-ASPP2), resulting ASPP2 gene over-expression. The apoptosis, autophagy and cell cycle changes were induced by culturing with serum-free medium for 24 h. Apoptosis was evaluated by Calcein／PI uptaking test, and autophagy was observed by counting the red fluorescent protein autophagy plasmid CFP-Lc3 which was transfected into cytoplasm. Cell cycle was detected by flow cytometry. Statistical analysis was performed by one-way analysis of variance (ANOVA). Results Over-expressed ASPP2 was found to significantly promote starvation-induced HCT116 apoptosis（cell apoptosis rate: GFP+ starvation group : 10.00%±1.42%；ASPP2+ starvation group: 18.44%±2.06%  P=0.000） and autophagy（cell autophagy rate: GFP+ starvation group : 35.00%±5.34%；ASPP2+ starvation group : 57.61%±6.06%  P=0.000）, and accelerate HCT116 G2/M arrest under starvation. But over-expressed ASPP2 could result in both G0/G1 and G2/M arrest without starvation. Conclusion These results suggest that ASPP2 can promote HCT116 p53+/+ cell apoptosis and autophagy, and affect the cell cycle.

Key words: Apoptosis, Cell cycle, Autophagy, Apoptosis stimulating protein 2 of p53