title: Primary functional identification of gene TMSG-1.
abstract: TMSG-1 was a tumor metastasis-related gene identified using mRNA differential display, whose expression level was lower in cancer cell lines with higher metastatic potential and in tumor tissue with metastasis. TMSG-1 was transfected to prostate cancer cell line (PC-3M-1E8) with high metastatic potential to observe the effects of increased expression of TMSG-1 on V-ATPase activity, intracellular pH and cell apoptosis. Subcellular localization of the encoded protein of TMSG-1 was determined by using GFP. Results showed that there were no differences of V-ATPase activity among parental PC-3M-1E8 cell line, pcDNA3 transfectant and anti-TMSG-1 transfectant, whereas the V-ATPase activity was significantly higher in TMSG-1 transfectant than that in parental PC-3M-1E8 cell line, pcDNA3 transfectant and Anti-TMSG-1 transfectant (p<0.001). Intracellular pH (pHi) was detected by using the pH-dependent fluorescence probe BECEF. Results showed the pHi was significantly increased in TMSG-1 transfectant. Cell apoptosis assay demonstrated cell apoptosis was significantly higher in -1 transfectant (p<0.01) and BCL2 expression was down regulated. Subcellular localization of TMSG-1 protein showed TMSG-1 was a transmembrane protein, which predicted TMSG-1 protein was located in cytoplasm system, such as endoplasmic reticulum and mitochondrial. These results indicated TMSG-1 up regulation in prostate cancer cell line could promote V-ATPase activity, increase pHi and cell apoptosis, and inhibit the expression of BCL2.
authors.affiliation: Department of Pathology, Health Science Center, Peking University, 100083, Beijing, China.
authors.forename: Chunshu
authors.initials: C
authors.lastname: Ma
authors.forename: Junyu
authors.initials: J
authors.lastname: Ning
authors.forename: Jiangfeng
authors.initials: J
authors.lastname: You
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.forename: Jieliang
authors.initials: J
authors.lastname: Wang
authors.forename: Xianglin
authors.initials: X
authors.lastname: Cui
authors.forename: Bingquan
authors.initials: B
authors.lastname: Wu
authors.forename: Jie
authors.initials: J
authors.lastname: Zheng