Overexpression of human tumor metastasis-related gene TMSG-1 suppresses cell proliferation and invasion of a highly metastatic prostate cancer cell line PC-3M-1E8 in vitro

目的 研究肿瘤转移抑制基因1(TMSG-1)转染对高转移人前列腺癌细胞体外增殖能力和侵袭能力的影响.方法 将TMSG-1全长真核表达载体稳定转染人前列腺癌细胞高转移亚系PC-3M-1E8,G418筛选,逆转录聚合酶链反应(RT-PCR)和Western blot方法选取TMSG-1过表达阳性克隆.通过活细胞计数、二苯基溴化四氮唑蓝(MTF)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.结果 在稳定转染TMSG-1基因的PC-3M-1E8细胞系中,筛选出3株TMSG-1转录活性及蛋白表达水平均明显升高的细胞用于后续生物学行为实验.活细胞计数和MTT比色实验结果显示,从计数第3天起,与空载体对照组和空白对照组相比,3株正义转染组细胞生长速度均明显减慢(P<0.05).软琼脂集落形成实验结果显示,3株正义转染组的细胞软琼脂集落形成数与空载体对照组、空白对照组相比,均明显减少(P<0.05),分别为26.00±3.21、13.33±1.45和32.83±2.18.Matrigel穿膜实验结果显示,正义转染的各组细胞与空载体对照组及空白对照组相比,穿膜细胞数均明显减少(P<0.05),分别为45.33±4.16、54.00±2.83和26.33±3.79.结论 TMSG-1表达上调可使高转移人前列腺癌细胞体外增殖速度、锚着不依赖性生长能力及侵袭能力明显降低,TMSG-1可能通过抑制肿瘤细胞生长和侵袭抑制肿瘤转移潜能.Objective To investigate the effects of tumor metastasis-related gene TMSG-1overexpression on the proliferation and invasion of a highly metastatic prostate cancer cell line in vitro. Methods The eukaryotic expression plasmids containing full-length TMSG-1 cDNAs were stably transfected into the highly metastatic prostate cancer cell line PC-3M-1E8. Clones highly expressing TMSG-1 were identified by RT-PCR and Western Blot analysis after G418 screening. The cell proliferation was detected by cell growth curve, MTT assay and soft agar colony formation assay. The invasive potential of tumor cells in vitro was tested by Matrigel invasion assay. Results Three TMSG-1 overexpression clones were selected. Cell growth curve and MTT assay showed that TMSG-I overexpression clones exhibited a strong inhibition of proliferation compared with that of the parental cells or those transfected with vector alone from the third day of culture (P<0.05). In vitro analysis also showed that the TMSG-1 transfected clones exhibited a decreased clonogenicity in soft agar compared with that of the parental cells or those transfected with vector only (P<0.05). TMSG-1 expression significantly suppressed cell invasion in vitro of TMSG-l-transfected PC-3M-1E8 cells (P<0.05). Conclusion The TMSG-1 protein may serve as a tumor metastasis suppressor due to inhibiting cell proliferation and invasion of the highly metastatic prostate cancer cell linePC-3M-1E8.国家自然科学基金; 国家重点基础研究发展计划(973计划); 教育部科学技术研究重点项目; 国家科技攻关计划中文核心期刊要目总览(PKU)中国科技核心期刊(ISTIC)中国科学引文数据库(CSCD)06404-4073