Abstract:
BACKGROUND:Mutation or deletion in the phosphatase and tensin homologue deleted on chromosome ten (PTEN) gene has been identified as an important cause of endometrial carcinoma; stromal cell derived factor-1alpha (SDF-1alpha) exerts growth-promoting effects on endometrial cancer cells through activation of the PI-3 kinase/Akt pathway and downstream effectors such as extracellular-responsive kinase (ERK). In this study, a plasmid containing the PTEN gene was transfected into Ishikawa cells to investigate the difference in growth and signal transduction between Ishikawa-PTEN and Ishikawa cells after SDF-1alpha stimulation, and to study mechanisms of the involvement of PTEN protein in endometrial carcinoma development. METHODS:Ishikawa cells were transfected with a plasmid (pLXSN-PTEN) containing the PTEN gene and a plasmid (pLXSN-EGFP) with enhanced green fluorescent protein (EGFP). Cells were then screened to obtain Ishikawa-PTEN cells and Ishikawa-neo cells that can both stably express PTEN protein and EGFP. Expression of PTEN protein, phosphorylation levels of AKT and ERK (pAKT and pERK) and growth differences in Ishikawa-PTEN, Ishikawa-neo and Ishikawa cells before and after SDF-1alpha stimulation were then determined by Western blots and MTT assays. RESULTS:Western blot analysis showed that Ishikawa cells produced PTEN after transfection with the PTEN gene. At 15 minutes after SDF-1alpha stimulation, the pAKT level of Ishikawa-PTEN cells was lower than that of Ishikawa-neo cells and Ishikawa cells. There was no significant difference in pERK levels among the three cell lines. The positive effect of SDF-1alpha on Ishikawa-PTEN cells growth was markedly less than the effect on Ishikawa-neo and Ishikawa cells. However, in the absence of SDF-1alpha stimulation (baseline), the pAKT level in Ishikawa-PTEN cells was less than that in Ishikawa cells. There was a significant difference in growth between the Ishikawa-PTEN cells and the Ishikawa-neo cells. CONCLUSIONS:PTEN gene transfection can regulate the level of pAKT but not pERK in Ishikawa-PTEN cells. PTEN protein may suppress the growth-promoting effect of SDF-1alpha on endometrial carcinoma by inhibiting the PI-3K/AKT signal transduction pathway.
journal_name
Chin Med J (Engl)journal_title
Chinese medical journalauthors
Li XP,Zhao D,Gao M,Zhao C,Wang JL,Wei LHkeywords:
subject
Has Abstractpub_date
2006-03-05 00:00:00pages
378-83issue
5eissn
0366-6999issn
2542-5641journal_volume
119pub_type
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journal_title:Chinese medical journal
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pub_type: 信件
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journal_title:Chinese medical journal
pub_type: 杂志文章
doi:10.4103/0366-6999.198932
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doi:
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journal_title:Chinese medical journal
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