EFFECT OF miR-21, miR-182 AND let-7i ON TSP-1 EXPRESSION IN COLON CANCER CELL LINE

Risultato della ricerca: Book/Film/Article review

Abstract

Background:MicroRNAs (miRNAs) are small non-coding RNAsthat regulate the expression of different genes, including genesinvolved in cancer progression, angiogenesis and metastasis.Thrombospondin-1 (TSP-1) has been shown to contrastangiogenesis in vivo. TSP-1 expression levels are inverselycorrelated with tumor vascularity and metastasis in coloncancer. Bio-informatic statistical analysis indicated that TSP-1 ishypothetical target of miR-21, miR-182, overexpressed in CRC, andlet-7i which expression is down-regulated in this tumor. In thiswork we investigated whether TSP-1 expression could be regulatedby miR-21, mir-182 and let-7i in HT29 colon cancer cell line.Methods:To investigated whether miR-21, mir-182 and let-7idirectly modulates TSP-1 expression, we transfected HT29 cell linewith pre-mir21, pre-mir182 and pre-let7i by using siPortNeo FXtranfection agent and after 48h we evaluated TSP-1 mRNA, usingQuantitative Real Time-PCR, and intracellular and secreted proteinlevel performed by Western blotting and ELISA. To confirm themodulation of TSP-1 by miRNAs we transfected HT29 cell line withanti-mir to target the mature form of miR-21, miR182 and let-7i.Results:Using Real-Time PCR we did not find any variation ofTSP-1 mRNA expression levels after transfection with pre-mir21 inHT29 cell line, but we observed a down-regulation of cytosolic andsecreted protein by Western blot and ELISA. In cells transfectedwith pre-mir182 we did not observe any down-regulation both TSP-1 mRNA and cytosolic and secreted protein. Finally, we didnot find any variation of TSP-1 level in cells transfected with let-7i. Results were confirmed by transfection with anti-mir21, anti-mir182 and anti-let7i and, using the same method, we evaluatedTSP-1 expression.Conclusions:Data suggest that mir-182 induces degradation ofTSP-1 mRNA in HT29 cell line, whereas mir-21 affects probablyby blockage of TSP-1 translation. Let-7i does not seem involved inregulation of TSP-1 expression in HT29 cells. Understanding themolecular mechanism by which miRNAs regulate TSP-1 expressioncould be used to restore TSP-1 expression to contrast angiogenicevents in colon cancer.
Lingua originaleEnglish
pagine (da-a)104-105
Numero di pagine1
RivistaCancer Treatment Reviews
Volume36
Stato di pubblicazionePublished - 2010

Fingerprint

Thrombospondin 1
Colonic Neoplasms
Cell Line
HT29 Cells
MicroRNAs
Messenger RNA
Transfection
Real-Time Polymerase Chain Reaction
Down-Regulation
Western Blotting
Enzyme-Linked Immunosorbent Assay
Neoplasm Metastasis
Neoplasms
Computational Biology

Cita questo

@article{b39af9f1b27045e1a25d02283052b95c,
title = "EFFECT OF miR-21, miR-182 AND let-7i ON TSP-1 EXPRESSION IN COLON CANCER CELL LINE",
abstract = "Background:MicroRNAs (miRNAs) are small non-coding RNAsthat regulate the expression of different genes, including genesinvolved in cancer progression, angiogenesis and metastasis.Thrombospondin-1 (TSP-1) has been shown to contrastangiogenesis in vivo. TSP-1 expression levels are inverselycorrelated with tumor vascularity and metastasis in coloncancer. Bio-informatic statistical analysis indicated that TSP-1 ishypothetical target of miR-21, miR-182, overexpressed in CRC, andlet-7i which expression is down-regulated in this tumor. In thiswork we investigated whether TSP-1 expression could be regulatedby miR-21, mir-182 and let-7i in HT29 colon cancer cell line.Methods:To investigated whether miR-21, mir-182 and let-7idirectly modulates TSP-1 expression, we transfected HT29 cell linewith pre-mir21, pre-mir182 and pre-let7i by using siPortNeo FXtranfection agent and after 48h we evaluated TSP-1 mRNA, usingQuantitative Real Time-PCR, and intracellular and secreted proteinlevel performed by Western blotting and ELISA. To confirm themodulation of TSP-1 by miRNAs we transfected HT29 cell line withanti-mir to target the mature form of miR-21, miR182 and let-7i.Results:Using Real-Time PCR we did not find any variation ofTSP-1 mRNA expression levels after transfection with pre-mir21 inHT29 cell line, but we observed a down-regulation of cytosolic andsecreted protein by Western blot and ELISA. In cells transfectedwith pre-mir182 we did not observe any down-regulation both TSP-1 mRNA and cytosolic and secreted protein. Finally, we didnot find any variation of TSP-1 level in cells transfected with let-7i. Results were confirmed by transfection with anti-mir21, anti-mir182 and anti-let7i and, using the same method, we evaluatedTSP-1 expression.Conclusions:Data suggest that mir-182 induces degradation ofTSP-1 mRNA in HT29 cell line, whereas mir-21 affects probablyby blockage of TSP-1 translation. Let-7i does not seem involved inregulation of TSP-1 expression in HT29 cells. Understanding themolecular mechanism by which miRNAs regulate TSP-1 expressioncould be used to restore TSP-1 expression to contrast angiogenicevents in colon cancer.",
author = "Daniele Fanale and {La Paglia}, Laura and Valeria Amodeo and Marta Castiglia and Naomi Margarese and Viviana Bazan and Antonio Russo and Marianna Terrasi and {Di Piazza}, Florinda and Lavinia Insalaco and Corsini, {Lidia Rita}",
year = "2010",
language = "English",
volume = "36",
pages = "104--105",
journal = "Cancer Treatment Reviews",
issn = "0305-7372",
publisher = "W.B. Saunders Ltd",

}

TY - JOUR

T1 - EFFECT OF miR-21, miR-182 AND let-7i ON TSP-1 EXPRESSION IN COLON CANCER CELL LINE

AU - Fanale, Daniele

AU - La Paglia, Laura

AU - Amodeo, Valeria

AU - Castiglia, Marta

AU - Margarese, Naomi

AU - Bazan, Viviana

AU - Russo, Antonio

AU - Terrasi, Marianna

AU - Di Piazza, Florinda

AU - Insalaco, Lavinia

AU - Corsini, Lidia Rita

PY - 2010

Y1 - 2010

N2 - Background:MicroRNAs (miRNAs) are small non-coding RNAsthat regulate the expression of different genes, including genesinvolved in cancer progression, angiogenesis and metastasis.Thrombospondin-1 (TSP-1) has been shown to contrastangiogenesis in vivo. TSP-1 expression levels are inverselycorrelated with tumor vascularity and metastasis in coloncancer. Bio-informatic statistical analysis indicated that TSP-1 ishypothetical target of miR-21, miR-182, overexpressed in CRC, andlet-7i which expression is down-regulated in this tumor. In thiswork we investigated whether TSP-1 expression could be regulatedby miR-21, mir-182 and let-7i in HT29 colon cancer cell line.Methods:To investigated whether miR-21, mir-182 and let-7idirectly modulates TSP-1 expression, we transfected HT29 cell linewith pre-mir21, pre-mir182 and pre-let7i by using siPortNeo FXtranfection agent and after 48h we evaluated TSP-1 mRNA, usingQuantitative Real Time-PCR, and intracellular and secreted proteinlevel performed by Western blotting and ELISA. To confirm themodulation of TSP-1 by miRNAs we transfected HT29 cell line withanti-mir to target the mature form of miR-21, miR182 and let-7i.Results:Using Real-Time PCR we did not find any variation ofTSP-1 mRNA expression levels after transfection with pre-mir21 inHT29 cell line, but we observed a down-regulation of cytosolic andsecreted protein by Western blot and ELISA. In cells transfectedwith pre-mir182 we did not observe any down-regulation both TSP-1 mRNA and cytosolic and secreted protein. Finally, we didnot find any variation of TSP-1 level in cells transfected with let-7i. Results were confirmed by transfection with anti-mir21, anti-mir182 and anti-let7i and, using the same method, we evaluatedTSP-1 expression.Conclusions:Data suggest that mir-182 induces degradation ofTSP-1 mRNA in HT29 cell line, whereas mir-21 affects probablyby blockage of TSP-1 translation. Let-7i does not seem involved inregulation of TSP-1 expression in HT29 cells. Understanding themolecular mechanism by which miRNAs regulate TSP-1 expressioncould be used to restore TSP-1 expression to contrast angiogenicevents in colon cancer.

AB - Background:MicroRNAs (miRNAs) are small non-coding RNAsthat regulate the expression of different genes, including genesinvolved in cancer progression, angiogenesis and metastasis.Thrombospondin-1 (TSP-1) has been shown to contrastangiogenesis in vivo. TSP-1 expression levels are inverselycorrelated with tumor vascularity and metastasis in coloncancer. Bio-informatic statistical analysis indicated that TSP-1 ishypothetical target of miR-21, miR-182, overexpressed in CRC, andlet-7i which expression is down-regulated in this tumor. In thiswork we investigated whether TSP-1 expression could be regulatedby miR-21, mir-182 and let-7i in HT29 colon cancer cell line.Methods:To investigated whether miR-21, mir-182 and let-7idirectly modulates TSP-1 expression, we transfected HT29 cell linewith pre-mir21, pre-mir182 and pre-let7i by using siPortNeo FXtranfection agent and after 48h we evaluated TSP-1 mRNA, usingQuantitative Real Time-PCR, and intracellular and secreted proteinlevel performed by Western blotting and ELISA. To confirm themodulation of TSP-1 by miRNAs we transfected HT29 cell line withanti-mir to target the mature form of miR-21, miR182 and let-7i.Results:Using Real-Time PCR we did not find any variation ofTSP-1 mRNA expression levels after transfection with pre-mir21 inHT29 cell line, but we observed a down-regulation of cytosolic andsecreted protein by Western blot and ELISA. In cells transfectedwith pre-mir182 we did not observe any down-regulation both TSP-1 mRNA and cytosolic and secreted protein. Finally, we didnot find any variation of TSP-1 level in cells transfected with let-7i. Results were confirmed by transfection with anti-mir21, anti-mir182 and anti-let7i and, using the same method, we evaluatedTSP-1 expression.Conclusions:Data suggest that mir-182 induces degradation ofTSP-1 mRNA in HT29 cell line, whereas mir-21 affects probablyby blockage of TSP-1 translation. Let-7i does not seem involved inregulation of TSP-1 expression in HT29 cells. Understanding themolecular mechanism by which miRNAs regulate TSP-1 expressioncould be used to restore TSP-1 expression to contrast angiogenicevents in colon cancer.

UR - http://hdl.handle.net/10447/104201

M3 - Book/Film/Article review

VL - 36

SP - 104

EP - 105

JO - Cancer Treatment Reviews

JF - Cancer Treatment Reviews

SN - 0305-7372

ER -