[Effect of phenethyl isothiocyanate on expression of genes during differentiation process of mouse embryonic stem cells in vitro].
OBJECTIVE
To study the effect of phenethyl isothiocyanate on the expression of protein kinase C alpha and the developmental genes (pax-6 and netrin-1) during differentiation process of mouse embryonic stem cells into neuron-like cells in vitro.


METHODS
Mouse embryonic stem cells were collected after 7 days in the presence of 1 x 10(-6) mol/L retinoic acid (RA), these special genes (nestin, glutaminase, Brn-3) of the neuron-like cells were measured by RT-PCR assay. Embryonic stem cells were cultured in the presence of retinoic acid (RA) and under the different concentrations of PEITC. These cultures were collected after 1, 3, 5, 7 and 14 days and mRNA levels of PKC, pax-6 and netrin-1 in undifferentiated and differentiated mouse embryonic stem cells were assessed by RT-PCR.


RESULTS
Large amounts of neurospheres were derived from mouse embryonic stem cells in the selected medium on the 7th day, which could express nestin, glutaminase and Brn-3 genes. RT-PCR assay showed that levels of PKC alpha, pax-6 and netrin-1 transcripts of the neuron-like cells decreased dramatically upon induction on the 1st day, but then raised slowly and resumed to normal level on 14 th day. PEITC down regulated the levels of PKC alpha, pax-6 and netrin-1 during various stages, also with the function concentrations of PEITC increasing, the tendency of drop to be obvious.


CONCLUSIONS
Phenethyl isothiocyanate has certain interference effects on the expression of developmental genes (pax-6, netrin-1) during the process of the ESCs' differentiation through suppressing PKC alpha expression.
Yu
Zhou
Z
Institute of Nutrition and Food Safety, China CDC, Beijing.
Xu
Haibin
H
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology*; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D017879:Isothiocyanates / Q000494:pharmacology*; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics / Q000378:metabolism; D000075388:Netrin-1; D009474:Neurons / Q000166:cytology*; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism; D051571:Protein Kinase C-alpha / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
D005136:Eye Proteins; D018398:Homeodomain Proteins; D017879:Isothiocyanates; D009414:Nerve Growth Factors; C000617353:Ntn1 protein, mouse; D000071841:PAX6 Transcription Factor; D051761:Paired Box Transcription Factors; C000605732:Pax6 protein, mouse; D012333:RNA, Messenger; D012097:Repressor Proteins; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1; C058305:phenethyl isothiocyanate; D051571:Protein Kinase C-alpha