Spindlin1, a novel nuclear protein with a role in the transformation of NIH3T3 cells.
spindlin1, a novel human gene recently isolated by our laboratory, is highly homologous to mouse spindlin gene. In this study, we cloned cDNA full-length of this novel gene and send it to GenBank database as spindlin1 (Homo sapiens spindlin1) with Accession No. AF317228. In order to investigate the function of spindlin1, we studied further the subcellular localization of Spindlin1 protein and the effects of spindlin1 overexpression in NIH3T3 cells. The results showed that the fusion protein pEGFP-N1-spindlin1 was located in the nucleus and the C-terminal is correlated with nuclear localization of Spindlin1 protein. NIH3T3 cells which could stably express spindlin1 as a result of RT-PCR analysis compared with the control cells displayed a complete morphological change; made cell growth faster; and increased the percentage of cells in G2/M and S phase. Furthermore, overexpressed spindlin1 cells formed colonies in soft agar in vitro and formed tumors in nude mice. Our findings provide direct evidence that spindlin1 gene may contribute to tumorigenesis.
Gao
Yanhong
Y
Laboratory of Stem Cell Biology, Beijing Institute of Transfusion Medicine, 27 Taiping Road, Beijing 100850, PR China.
Yue
Wen
W
Zhang
Peng
P
Li
Li
L
Xie
Xiaoyan
X
Yuan
Hongfeng
H
Chen
Lin
L
Liu
Daqing
D
Yan
Fang
F
Pei
Xuetao
X
D000362:Agar / Q000737:chemistry; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019556:COS Cells; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000096:biosynthesis* / Q000502:physiology*; D002455:Cell Division; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005091:Exons; D020224:Expressed Sequence Tags; D005260:Female; D005434:Flow Cytometry; D016195:G2 Phase; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D007438:Introns; D051379:Mice; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis* / Q000502:physiology*; D008969:Molecular Sequence Data; D041681:NIH 3T3 Cells; D016366:Open Reading Frames; D010051:Ovarian Neoplasms / Q000473:pathology; D010750:Phosphoproteins / Q000096:biosynthesis* / Q000502:physiology*; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase; D014162:Transfection
D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D008869:Microtubule-Associated Proteins; D010750:Phosphoproteins; C104434:spindlin; D049452:Green Fluorescent Proteins; D000362:Agar