Abstract | BACKGROUND: RESULTS: This study documents the molecular presence of human chorionic gonadotropin beta subunit in uterine cervix cancer tissues and investigates a novel technique to reduce hCGbeta levels based on expression of a modified U1 snRNA as a method to study the hormone's role in biology of human cervical cancer cells cultured in vitro. The property of U1 snRNA to block the accumulation of specific RNA transcript when it binds to its donor sequence within the 3' terminal exon was used. The first 10 nucleotides of the human U1 snRNA gene, which normally binds to the 5'ss in pre-mRNA were replaced by a sequence complementary to a 10-nt segment in the terminal exon of the hCGbeta mRNA. Three different 5' end-mutated U1 snRNA expression plasmids were tested, each targeting a different sequence in the hCGbeta mRNA, and we found each one blocked the expression of hCGbeta in HeLa cells, a cervix carcinoma cell line, as shown by immunohistochemistry and qRT-PCR. Reduction of hCGbeta levels resulted in a significantly increased apoptosis rate with almost 90% of cells transfected with modified anti-hCGbeta U1 snRNAs showing morphological changes characteristic of the apoptotic process. CONCLUSION:
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Authors | Anna Jankowska, Samuel I Gunderson, Miroslaw Andrusiewicz, Beata Burczynska, Anna Szczerba, Artur Jarmolowski, Ewa Nowak-Markwitz, Jerzy B Warchol |
Journal | Molecular cancer
(Mol Cancer)
Vol. 7
Pg. 26
(Mar 14 2008)
ISSN: 1476-4598 [Electronic] England |
PMID | 18339208
(Publication Type: Journal Article, Research Support, N.I.H., Extramural)
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Chemical References |
- Chorionic Gonadotropin, beta Subunit, Human
- RNA, Messenger
- RNA, Small Nuclear
- U1 small nuclear RNA
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Topics |
- Apoptosis
- Cell Cycle
- Chorionic Gonadotropin, beta Subunit, Human
(antagonists & inhibitors, genetics, metabolism)
- Female
- Gene Expression Regulation, Neoplastic
- Gene Silencing
- HeLa Cells
- Humans
- Immunohistochemistry
- RNA, Messenger
(genetics, metabolism)
- RNA, Small Nuclear
(metabolism)
- Transfection
- Uterine Cervical Neoplasms
(genetics, pathology)
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