HAP1对乳腺癌细胞MCF-7生物学特性影响的观察  

Effect of HAP1gene overexpression on biological characteristics of breast cancer cell MCF-7

作　　者：宋雪[1] 朱丽伟[1] 唐金海[1] 吴建中[2] 马蓉[2] 曹海霞[2] 

机构地区：[1]南京医科大学附属江苏省肿瘤医院普外科,江苏南京210000 [2]南京医科大学附属江苏省肿瘤医院临床肿瘤研究中心,江苏南京210000

出　　处：《中华肿瘤防治杂志》2013年第7期497-501,共5页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:探讨亨廷顿相关蛋白1(HAP1)基因过表达对人乳腺癌细胞株MCF-7增殖、体外迁移侵袭和细胞凋亡的影响及其可能机制。方法:通过转染的方法将逆转录病毒pBabe-puro(嘌呤霉素)HAP1质粒和pBabe-puro质粒导入人乳腺癌细胞系MCF-7,用嘌呤霉素筛选稳定表达两质粒的细胞系,荧光定量PCR和蛋白质印迹法鉴定是否成功构建HAP1过表达细胞系;细胞增殖-毒性检测试剂盒(CCK-8)和克隆形成实验检测细胞的生长增殖,Transwell小室法检测细胞的侵袭和迁移,流式细胞仪检测细胞的凋亡。结果:成功构建稳定表达pBabe-HAP1的MCF-7-pBabe-puro-HAP1细胞模型。CCK-8检测72h细胞增殖率,MCF-7-pBabe-puro-HAP1为(75.97±6.76)%,明显低于MCF-7-pBabe-puro细胞(93.98±6.63)%(P=0.03)及MCF-7细胞(100.00±0.00)%,P=0.004;MCF-7-pBabe-puro-HAP1细胞克隆形成率为(22.67±1.26)%,明显低于MCF-7(35.00±0.50)%(P=0.000)和MCF-7-pBabe-puro细胞(33.83±0.76)%,P=0.000;Transwell小室侵袭和迁移实验表明,MCF-7-pBabe-puro-HAP1组的侵袭(3.33±0.58,P=0.000)和迁移(50.00±3.61,P<0.01)能力明显降低;流式细胞仪检测细胞凋亡,MCF-7-pBabe-puro-HAP1凋亡率为(8.03±0.15)%,高于MCF-7-pBabe-puro(3.13±0.25)%(P=0.000)和MCF-7细胞(3.33±0.35)%,P=0.000。结论:HAP1基因能够抑制肿瘤细胞增殖和迁移侵袭,并能诱导细胞凋亡,其可能作为一个抑癌基因在肿瘤发生发展中发挥重要作用。OBJECTIVE： To investigate the effect of HAP1 gene overexpression on proliferation,migration and invasion capability,cell apoptosis of MCF-7 breast cancer cell line in vitro and possible mechanism. METHODS： Human breast cancer cell line MCF-7 was cultured and transfected with recombinant plasmid pBabe-puro-HAP1 or blank plasmid pBabe- puro. Real-time PCR and Western blot were used to detect the mRNA and protein expression of HAP1. The cell proliferation was detected by CCK-8 assay. The migration and invasion capability in vitro was detected by Transwell chamber as say. Clone formation assay was carried out to determine the clonogenicity. Cell apoptosis was determined by flow cytome- try. RESULTS：The cell model MCF-7-pBabe-puro-HAP1 was successfully constructed, which was stably expressing pB- abe-HAP1. After 72 h cuhuring,CCK-8 assay showed that the proliferation rate of MCF-7-pBabe-puro-HAP1 group was （75.97＋6. 76）%,which was higher than those of MCF-7-pBabe-puro （93. 98±6. 63）% （P=0. 03） and MCF-7 （100.00±0.00） %（P=0. 004）groups. The abilities of migration（50.00±3.61 ,P〈0.01） and invasion（3.33±0.58,P= 0. 000） of the MCF-7-pBabe-puro-HAP1 cells were obviously decreased as compared with those MCF-7 and MCF-7-pB- abe-puro cells. The apoptosis rate of MCF-7-pBabe-puro-HAP1 cells was （8.03 ±0. 15）%, which was also higher than those of MCF-7-pBabe-puro（3.13±0. 25） %（P=0. 000）and MCF-7（3.33±0. 35）%（P=0. 000） cells. The cloning effi- ciency of MCF-7-pBabe-puro-HAP1 was （22.67±1.26）%,which was lower than those of MCF-7（35. 00±0.50）%（P= 0. 000） and MCF-7-pBabe-puro（33.83 ± 0.76） % （P = 0. 000）. CONCLUSIONS： HAP1 gene overexpression can inhibit the proliferation,migration and invasion capabilities,and induce the apoptosis of MCF-7 in vitro. It may play an important role in biological function as a tumor suppressor gene.

关 键 词：乳腺肿瘤 亨廷顿相关蛋白1 细胞凋亡 MCF-7 

分 类 号：R737.9[医药卫生—肿瘤]