Comparative transcript profiles of cell cycle-related genes in mouse primordial germ cells, embryonic stem cells and embryonic germ cells.
We used cDNA array to compare the relative transcript levels of 96 cell cycle-related genes in mouse primordial germ cells (PGCs), embryonic germ (EG) cells and embryonic stem (ES) cells. Among 38 genes of the G1 phase analysed, Ccnd3 (CyclinD3), Cdkn1c (p57(kip2)), Rb1, and Tceb1l (Skip1-like) were expressed at significantly higher levels in PGCs than in EG and ES cells; Ccnd1 (CyclinD1) was more abundant in EG cells than in PGCs. Except for higher mRNA levels of Ccng (CyclinG1) in EG and ES cells in comparison to PGCs, no difference among 20 genes of the S and 12 genes of G2/M phases was found. Less than half of the 26 genes regarded as DNA damage checkpoint/Trp53/Atm pathway genes showed significant transcript levels in all three cell populations. Among these, the transcript levels of Ube1x and Atm were significantly higher in PGCs than in EG and ES cells while that of Ube3a was higher in these latter. In addition, relatively high mRNA levels of Timp3 characterizes EG cells while transcripts of this gene were very low in PGCs and barely detectable in ES cells. With the exception of Tceb1l, differential transcript levels found in the cDNA array assay were confirmed by real time RT-PCR. Using this method, we also analysed the transcripts of two genes not present in the cDNA array: c-myc, known to be critical for the control of cell cycle in many cell types, and Eras, specifically expressed in ES cells and involved in the control of ES cell proliferation and their tumorigenic properties. While c-myc transcripts were present at similar levels in all three cell types examined, Eras was expressed at high levels in ES cells (10-fold) and even more so in EG cells (almost 40-fold) in comparison to PGCs. Taken together, these results indicate that despite similarities between PGCs and ES or EG cells, their cell cycles are differently regulated. In particular, it appears that PGCs, like most mitotic cells, possess a more regulatable control of G1 phase than EG and ES cells. Moreover, our data provide useful clues for further studies aimed at identifying cell cycle genes critical for PGC growth and their transformation in tumorigenic cells.
Sorrentino
Eleonora
E
Department of Public Health and Cell Biology, University of Rome Tor Vergata, Rome, Italy.
Nazzicone
Valeria
V
Farini
Donatella
D
Campagnolo
Luisa
L
De Felici
Massimo
M
D000818:Animals; D002453:Cell Cycle / Q000235:genetics*; D002455:Cell Division / Q000235:genetics / Q000502:physiology; D056743:Cyclin D3; D050761:Cyclin-Dependent Kinase Inhibitor p57 / Q000235:genetics; D016213:Cyclins / Q000235:genetics; D000075065:Elongin; D053595:Embryonic Stem Cells / Q000502:physiology*; D016193:G1 Phase / Q000235:genetics / Q000502:physiology; D016195:G2 Phase / Q000235:genetics / Q000502:physiology; D020869:Gene Expression Profiling; D005854:Germ Cells / Q000502:physiology*; D051379:Mice; D016160:Retinoblastoma Protein / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic
C534697:Ccnd3 protein, mouse; C493965:Cdkn1c protein, mouse; D056743:Cyclin D3; D050761:Cyclin-Dependent Kinase Inhibitor p57; D016213:Cyclins; D000075065:Elongin; D016160:Retinoblastoma Protein; D014157:Transcription Factors