组蛋白去乙酰化酶1在人肺癌细胞株A549中的表达及其对裸鼠移植瘤放射敏感性的影响  

Expression of histone deacetylase 1 gene in human lung cancer cell line A549 and the effect on the radiation sensitivity of nude mouse transplantation tumor

作　　者：郭佳[1] 何彬[2] 黄际远[1] 宋文忠[1] 张明[3] 

机构地区：[1]四川省人民医院核医学科,成都610072 [2]四川省人民医院胸外科,成都610072 [3]四川省人民医院肿瘤科,成都610072

出　　处：《中华实验外科杂志》2017年第11期1931-1933,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨组蛋白去乙酰化酶1（HDAC1）基因在人肺癌A549细胞中的表达，并通过构建裸鼠移植瘤检测其对裸鼠放射敏感性的影响。方法将HDAC1小干扰RNA（siRNA）和重组表达载体pcDNA3.1-HDAC1分别转染肺癌A549细胞后，G418筛选得到稳定转染细胞株。反转录-聚合酶链反应（RT-PCR）和Western blot分别检测细胞中HDAC1 mRNA和蛋白表达水平；细胞计数试剂盒（CCK-8）检测细胞增殖，克隆形成实验观察上调和下调HDAC1表达对A549细胞放射敏感性的影响；原位缺口末端标记法（TUNEL）法检测细胞凋亡影响；裸鼠移植瘤实验检测上调和下调HDAC1表达联合X射线照射对肺癌的生长抑制作用。 结果HDAC1下调组肺癌A549细胞内HDAC1基因和蛋白表达分别降低为1.86±0.09和2.41±0.10（P＝0.015、0.006），HDAC1上调组明显升高至3.65±0.14和5.41±0.12（P＝0.012、0.008）；HDAC1下调组肺癌A549细胞放射敏感性升高（P＝0.025），HDAC1上调组肺癌A549细胞放射敏感性降低（P＝0.009）；HDAC1下调组肺癌A549细胞增殖被抑制，细胞凋亡率增加至（42.17±3.19）%（P＝0.006）；HDAC1上调组A549细胞增殖能力明显增强，细胞凋亡率降低至（9.11±1.46）%（P＝0.008），HDAC1下调组裸鼠移植瘤平均体积[（547.37±89.68） mm3]明显小于对照组[（1 238.44±212.36） mm3 ，P＝0.006]；HDAC1上调组的瘤体平均体积[（1 681.27±195.37） mm3]大于对照组（P＝0.011）。20 Gy γ射线照射后HDAC1下调组瘤体平均体积[（214.15±54.26） mm3]较照射前明显减小（P＝0.007）；而上调组裸鼠皮下种植瘤平均体积较照射前比较差异无统计学意义（P＝0.264）。结论下调HDAC1表达能抑制人肺癌A549细胞裸鼠移植瘤的生长，增强瘤体的放射敏感性，上调HDAC1表达则使肿瘤辐射抗拒。Objective To investigate the expression of histone deacetylase 1 （ HDAC1 ） gene in hu- man lung cancer cell line A549 and the effect on the radiation sensitivity of nude mouse transplantation tumor. Methods HDAC1 small interfering RNA （siRNA） and recombinant expression vector pcDNA3. 1 - HDAC1 were transfected with A549 lung cancer cells, respectively. Stable transfected cell lines were screened by G418 selection, reverse transcriptase - polymerase chain reaction （ RT - PCR） and Western blotting were used to detect the expression levels of HDAC1 mRNA and HDAC1 protein in the cell; cell proliferation was measured by cell counting kit - 8 （ CCK - 8 ）. Clone formation experiment was used to ob- serve the effect of up - regulation and down - regulation of HDAC1 expression on the radiation sensitivity of A549 cells, cell apoptosis was detected by transferase - mediated dUTP nick - end labelling （TUNEL） method. Nude mice experiment of transplantable tumor was applied for in vivo detection of the influence of up - regulated and down - regulation of HDAC1 expression combined with X - ray irradiation in the inhibi- tion of lung cancer growth. Results The expression of HDAC1 gene and protein were significantly lower to 1.86 ±0. 09 and 2.41 ±0. 10 （P =0. 015,0. 006） in HDAC1 cell line A549, and the HDAC1 up - regu- lation group were significantly higher to 3.65 ±0. 14 and 5.41 ±0. 12 （P =0. 012, 0. 008） ; The radiation sensitivity of lung cancer A549 cells was increased in the down - regulation group of HDAC1 （P = 0. 025 ）, and the radiation sensitivity of lung cancer A549 cells was decreased （P = 0. 009 ） in the HDAC1 up - regu- lation group. Down- regulation of HDAC1 inhibited the proliferation of A549 lung cancer, increased the rate of apoptosis to （42. 17 ± 3.19） % （P = 0. 008 ）, proliferation of A549 cells was significantly enhanced in HDAC1 up - regulated group, associated with reduced percentage of apoptotic cells （9. 11 ± 1.46） % （P= 0.006）; Average volume in

关 键 词：肺癌 A549细胞 组蛋白去乙酰化酶1 移植瘤 裸鼠 

分 类 号：R734.2[医药卫生—肿瘤]