人肿瘤转移抑制基因1转染对人乳腺癌细胞MDA-MB-231体外生物学行为的影响  

Overexpression of tumor metastasis suppressor gene 1 suppresses proliferation and invasion,but enhances apoptosis of human breast cancer cells MDA-MB-231 cells

作　　者：苏静[1] 由江峰[1] 王洁良[1] 崔湘琳[1] 方伟岗[1] 郑杰[1] 

机构地区：[1]北京大学医学部病理学系,100083

出　　处：《中华病理学杂志》2007年第10期672-676,共5页Chinese Journal of Pathology

基　　金：国家自然科学基金(30170363);国家973重点基础研究发展规划资助项目(2002CB513100);教育部科学技术研究重大项目基金(01003);国家"十五"科技攻关资助项目(2001BA703805)

摘　　要：目的研究人肿瘤转移抑制基因1(TMSG-1)转染引起人乳腺癌细胞 MDA-MB-231体外生物学行为的改变及对肿瘤转移表型的影响。方法构建 TMSG-1全长编码序列真核表达载体,稳定转染人乳腺癌 MDA-MB-231细胞系,G418筛选挑取 TMSG-1过表达阳性克隆。通过 MTT 比色实验、软琼脂集落形成实验检测体外细胞生长能力;Matrigel 穿膜实验检测肿瘤细胞体外侵袭能力。TMSG-1瞬时转染24、48 h,分别用 Annexin-V 碘化丙啶(PI)双标流式细胞术检测肿瘤细胞凋亡情况。结果从稳定转染 TMSG-1的 MDA-MB-231细胞中挑取3个 TMSG-1-FLAG 融合蛋白表达量较高的阳性克隆株用于下游生物学行为实验。MTT 比色实验及软琼脂集落形成实验结果显示,TMSG-1正义转染各组(S1、S2、S3)细胞增殖速度及克隆形成数与未转染组及转染空载体组相比均明显减低(P<0.05);Matrigel 穿膜实验显示,正义转染各组的穿膜细胞数[(72.3±8.1)个、(85.0±4.2)个、(73.5±7.8)个]与未转染组[(187.5±2.1)个]和转染空载体组[(162.3±6.8)个]相比均明显减少(P<0.01)。TMSG-1瞬时转染 MDA-MB-231细胞,转染24和48 h 均可引起细胞凋亡率的增加(P<0.05)。结论 TMSG-1表达上调可使人乳腺癌细胞 MDA-MB-231体外生长速度、锚着不依赖性生长能力及侵袭能力明显降低,细胞凋亡增加。该实验为 TMSG-1是一个新发现的肿瘤转移抑制基因提供了证据。Objective To investigate the effects of tumor metastasis suppressor gene 1 （ TMSG-1 ） overexpression on the proliferation, invasion and apoptosis of breast cancer cells and to determine possible correlations of TMSG-1 and metastasis of breast cancer. Methods Full-length human TMSG-1 coding sequences were cloned into plasmid pcDNA3.0-FLAG. The recombinant plasmids constructs were transfeced into MDA-MB-231, a highly malignant breast cancer cell line. Parental, vector-only stable transfectant and TMSG-1 stable transfectant clones were tested by MTY, soft agar colony formation and Boyden chamber assays. At twenty-four hours and forty-eight hours post transient transfection, double staining with Annexin- V-FITC and PI were employed to distinguish apoptotic cells from living cells by flow cytometry analysis. Results Three TMSG-1 overexpression clones were selected. Compared with the control cells, TMSG-1 overexpression MDA-MB-231 cells showed strong inhibition of proliferation and decreased clonogenicity in soft agar（ P 〈 0. 05）. Transfection of TMSG-1 into MDA-MB-231 cells significantly suppressed the cell invasion ability in vitro （ decreased numbers of cells trespassing the matrigel in three experiments ： 72. 3 ± 8.1, 85. 0 ±4. 2, and 73.5±7. 8） in comparison with naive cells without transfection （187.5 ±2. 1 ） and cells transfected with the control vector （ 162. 3 ± 6. 8 ） （ P 〈 0. 01 ）. Transient transfection of TMSG-1 into MDA-MB-231 cells could promote cell apoptosis at 24 and 48 hours after transfection （ P 〈 0. 05 ）. Conclusions TMSG-1 protein may have multiple functions in the regulation of proliferation, invasion andapoptosis of metastatic breast cancer cells, likely as a metastasis suppressor gene.

关 键 词：乳腺肿瘤 肿瘤转移 转染 基因 TMSG-1 

分 类 号：R737.9[医药卫生—肿瘤]