Su Jing, You Jiang-feng, Wang Jie-liang, Cui Xiang-lin, Fang Wei-gang, Zheng Jie
Department of Pathology, Health Science Center, Peking University, Beijing 100083, China.
Zhonghua Zhong Liu Za Zhi. 2008 Jun;30(6):404-7.
To investigate the effects of tumor metastasis-related gene TMSG-1 overexpression on the proliferation and invasion of a highly metastatic prostate cancer cell line in vitro.
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.
Three TMSG-1 overexpression clones were selected. Cell growth curve and MTT assay showed that TMSG-1 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-1-transfected PC-3M-IE8 cells (P < 0.05).
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 line PC-3M-1E8.
探讨肿瘤转移相关基因TMSG-1过表达对高转移性前列腺癌细胞系体外增殖和侵袭的影响。
将含全长TMSG-1 cDNA的真核表达质粒稳定转染至高转移性前列腺癌细胞系PC-3M-1E8。经G418筛选后,通过RT-PCR和蛋白质免疫印迹分析鉴定高表达TMSG-1的克隆。采用细胞生长曲线、MTT法和软琼脂集落形成试验检测细胞增殖。通过基质胶侵袭试验检测肿瘤细胞的体外侵袭潜能。
筛选出3个TMSG-1过表达克隆。细胞生长曲线和MTT试验显示,与亲代细胞或仅转染载体的细胞相比,从培养第3天起,TMSG-1过表达克隆的增殖受到强烈抑制(P<0.05)。体外分析还显示,与亲代细胞或仅转染载体的细胞相比,转染TMSG-1的克隆在软琼脂中的克隆形成能力降低(P<0.05)。TMSG-1表达显著抑制了转染TMSG-1的PC-3M-IE8细胞的体外侵袭(P<0.05)。
TMSG-1蛋白可能通过抑制高转移性前列腺癌细胞系PC-3M-1E8的细胞增殖和侵袭而发挥肿瘤转移抑制作用。