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外源性PTEN增强放射线诱导胰腺癌ASPC-1细胞G2/M期阻滞
The block of exogenous PTEN on G2/M phase of pancreatic cancer cell lines ASPC-1 enhanced by radiation
【作者】 李建军; 陈延治; 李光; 顾菲; 王志宇; 辛彦;
【Author】 Li Jian-Jun Chen Yan-Zhi Li Guang Gu Fei Wang Zhi-Yu Tan Xin Department of Radiotherapy, The first affiliated Hospital, China Medical University, Shenyang, 110001, Liaoning, China Yan Xin No.4 Lab, Cancer Institute, The First Affiliated Hospital, China Medical University
【机构】 中国医科大学附属第一医院放疗科; 中国医科大学附属第一医院肿瘤研究所第四研究室;
【摘要】 目的:观察外源性PTEN在乏氧及放射前后对胰腺癌细胞系ASPC-1细胞周期及细胞存活的影响。方法:将质粒pEAK8和pEAK8-PTEN分别转染ASPC-1细胞,获得ASPC-1-Peak8细胞及高表达PTEN的ASPC-1- pEAK8-PTEN细胞,将ASPC-1-pEAK8-PTEN、ASPC- 1-pEAK8和ASPC-1细胞各分成常氧组、乏氧组、照射组、乏氧照射组。常氧组中,乏氧组施加乏氧(1%O2)处理, 乏氧时间为24 h,照射组接受4Gy单次照射,乏氧照射组在乏氧情况下进行照射。用流式细胞术、成克隆分析法检测外源性PTEN对ASPC-1细胞周期分布及细胞克隆形成能力的影响。结果:ASPC-1-pEAK8-PTEN细胞较ASPC- 1、ASPC-1-pEAK8细胞G2/M、S期细胞增多,G1期细胞减少,出现少量凋亡细胞。常氧情况下,ASPC-1、ASPC- 1-pEAK8和ASPC-1-pEAK8-PTEN细胞接受4Gy X线单次照射24 h后,ASPC-1、ASPC-1-pEAK8细胞G2/ M期细胞阻滞不明显,ASPC-1-pEAK8-PTEN细胞可以看到明显的G2/M期细胞阻滞。乏氧24 h后,ASPC-1- pEAK8-PTEN细胞G2/M期比例明显减少,重新出现高比例的G1期细胞。ASPC-1、ASPC-1-pEAK8细胞乏氧培养 24h后,仍以G1期细胞阻滞为主,并且G1期细胞与乏氧前比较进一步增加。ASPC-1、ASPC-1-pEAK8和ASPC- 1-pEAK8-PTEN细胞分别乏氧24h后在乏氧状态下进行 4Gy单次照射,照射24 h后,ASPC-1、ASPC-1-pEAK8 及ASPC-1-pEAK8-PTEN细胞在乏氧照射情况下看不到明显的G2/M期细胞阻滞,G1期细胞比例较高,凋亡细胞较少。常氧及放射后,ASPC-1、ASPC-1-pEAK8、ASPC- 1-pEAK8-PTEN细胞克隆形成率分别为33.33±8.38%、 31.67±4.32%、24.00±3.90%和5.53±.52%、5.33 ±.74%、3.73±.20%。乏氧及放射后,ASPC-1、ASPC- 1-pEAK8、ASPC-1-pEAK8-PTEN细胞克隆形成率分别为 29.67±4.97%、29.50±3.39%、19.83±5.12%和12.08 ±.78%、11.17±.73%和7.38±.58%。结果表明,常氧组ASPC-1-pEAK8-PTEN细胞克隆形成率较ASPC— 1,ASPC-1-pEAK8细胞降低明显(F=4.283,P<0.05), 常氧照射组ASPC-1-pEAK8-PTEN细胞克隆形成率较 ASPC-1、ASPC-1-pEAK8细胞降低更明显(F=7.766,P <0.01);乏氧组ASPC-1-pEAK8-PTEN细胞克隆形成率较 ASPC-1,ASPC-1-pEAK8细胞降低明显(F=9.152,P<0. 01),乏氧照射组ASPC-1-pEAK8-PTEN细胞克隆形成率较ASPC-1、ASPC-1-pEAK8细胞降低明显(F=75.802,P <0.01)。结论:外源性PTEN可使胰腺癌ASPC-1细胞阻滞在G2/M期,增强放射线诱导G2/M期阻滞的能力,提高放射线对ASPC-1细胞在常氧及乏氧情况下的杀伤能力。
【Abstract】 Objective: To investigate the effect of heteroenous Phosphatase and tensin homologue deleted on chromosome ten (PTEN) on cell cycles and plating efficiency (PE) in human pancreas cancer cell line ASPC-1 under hypoxia and radiation exposure conditions. Methods: After transfected with an eukaryotic expression plasmid (Peak8) containing PTEN or not in vitro by lipofectin, the ASPC-1 cell were named ASPC-l-pEAK8-PTEN, ASPC-l-pEAK8 or ASPC-1 cell respectively according to the contents of the cells. ASPC-l-pEAK8-PTEN, ASPC-l-pEAK8 and ASPC-1 cells were assigned to normal oxygen group, hypoxia group, radiation group and hypoxia radiation group respectively. Radiation groups and hypoxia radiation group (1%O2 for 24h) were treated with the dose of 4Gy X-ray. Flow cytometry and cologenic survival assay were adopted to measure the effects of PTEN on cell cycle and the plating efficiency of ASPC-1 cells respectively. Results: Under normal oxygen condition, the cell cycle analysis demonstrated an increase of the ASPC-1-pEAK8-PTEN cells in the G2/M and S phase whereas an decrease in G1 phase comparing to the ASPC-l,ASPC-1-pEAK8 cells. Twenty-foue hours after exposing to 4Gy X-rays, the G2/M phase block were not obviously in the ASPC-1 and ASPC-1-pEAK8 cells, whereas that in the ASPC-1-pEAK8-PTEN cells was obviously. But 24 h after exposing in hypoxia condition, all the three kinds of cells demonstrated G2/M phase block decrease obviously, whereas G1 phase increase obviously, and it demonstrated the same results when the radiation exposed under the hypoxia condition. In normal oxygen condition and radiation under that, the ASPC-1,ASPC-1-pEAK8, ASPC-1-pEAK8-PTEN cells plating efficiency were 33. 33 ± 8.38 %31.67 ± 4.32 %,24.00 ± 3.90 % and 5.53 ± . 52%,5.33 ± .74%,3.73 ± .20% respectively. In hypoxia condition and radiation under that, the ASPC-1,ASPC-1-pEAK8,ASPC-1-pEAK8-PTEN cells plating efficiency were 29.67 ± 4.97%,29.50 ± 3.39%,19.83 ± 5.12% 和 12. 08 ± .78%, 11.17 ± .73% and 738 ± .58% respectively. The results showed that in normal oxygen condition, the ASPC-1-pEAK8-PTEN cells plating efficiency was lower than that of ASPC-1,ASPC-1-pEAK8 cells(F=4.283, P<0.05), in radiation under normal oxygen condition, the ASPC-1-pEAK8-PTEN cells plating efficiency was much lower than that of ASPC-1,ASPC-1-pEAK8 cells (F=7.766, P <0.01),in hypoxia condition, the ASPC-1-pEAK8-PTEN cells plating efficiency was lower than that of ASPC-1, ASPC-1-pEAK8 cells (F=9.152, P<0.01), in radiation under hypoxia condition, the ASPC-1-pEAK8-PTEN cells plating efficiency was lower than that of ASPC-1 ,ASPC-1-pEAK8 cells (F=75.802, P<0.01). Conclusion: Exogenous PTEN can block ASPC-1 pancreatic cancer cells in G2/M phase, increase the ability of X-rays in G2/M phase blocking and decrease the plating efficiency of ASPC-1 cells under normal oxygen and hypoxia condition.
- 【会议录名称】 第四届中国肿瘤学术大会暨第五届海峡两岸肿瘤学术会议论文集
- 【会议名称】第四届中国肿瘤学术大会暨第五届海峡两岸肿瘤学术会议
- 【会议时间】2006-10
- 【会议地点】中国天津
- 【分类号】R735.9
- 【主办单位】中国抗癌协会、中华医学会肿瘤学分会