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Mxi1 inhibits the proliferation of U87 glioma cells through down-regulation of cyclin B1 gene expression.

Manni I, Tunici P, Cirenei N, Albarosa R, Colombo BM, Roz L, Sacchi A, Piaggio G, Finocchiaro G - Br. J. Cancer (2002)

Bottom Line: Interestingly, the expression of cyclin B1 was inhibited to about 60% in U87.Mxi1.14 cells.This inhibition occurs at the transcriptional level and depends, at least in part, on the E-box present on the cyclin B1 promoter.Consistent with this, the endogenous Mxi1 binds this E-box in vitro.

View Article: PubMed Central - PubMed

Affiliation: Istituto Regina Elena, Centro Ricerca Sperimentale, Laboratorio di Oncogenesi Molecolare, Via delle messi D'Oro 156, 00158 Rome, Italy.

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Mxi1 inhibits expression of the cyclin B1 gene. Northern blot analysis was performed on total RNA extracted from wild-type, Mxi1.22, and Mxi1.14 U87 cell lines. RNAs were size-fractionated on a 1% agarose gel, blotted on a nylon membrane, hybridized with 32P-labelled cyclin B1, cyclin A and MXI1 cDNAs and assessed by autoradiography. Exposure was for 3 h (after overnight exposure the endogenous MXI1 mRNA was visible). To normalize RNA loading, the membrane was re-hybridized with the 32P-labelled cDNA of the housekeeping gene β-actin. The relative intensity of the bands was quantitated by densitometry.
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fig5: Mxi1 inhibits expression of the cyclin B1 gene. Northern blot analysis was performed on total RNA extracted from wild-type, Mxi1.22, and Mxi1.14 U87 cell lines. RNAs were size-fractionated on a 1% agarose gel, blotted on a nylon membrane, hybridized with 32P-labelled cyclin B1, cyclin A and MXI1 cDNAs and assessed by autoradiography. Exposure was for 3 h (after overnight exposure the endogenous MXI1 mRNA was visible). To normalize RNA loading, the membrane was re-hybridized with the 32P-labelled cDNA of the housekeeping gene β-actin. The relative intensity of the bands was quantitated by densitometry.

Mentions: To dissect the molecular mechanism through which Mxi1 induces accumulation in the G2/M phase of the cell cycle, we investigated, in U87 proliferating cells, the expression of cyclin B1 gene and of cyclin A as control. The level of the two cyclin mRNAs was evaluated by Northern blot on RNA of U87.Mxi1.14, U87.Mxi1.22 and U87 wild-type cells. MXI1 and beta-actin transcripts were also analyzed on the same membrane. Figure 5Figure 5


Mxi1 inhibits the proliferation of U87 glioma cells through down-regulation of cyclin B1 gene expression.

Manni I, Tunici P, Cirenei N, Albarosa R, Colombo BM, Roz L, Sacchi A, Piaggio G, Finocchiaro G - Br. J. Cancer (2002)

Mxi1 inhibits expression of the cyclin B1 gene. Northern blot analysis was performed on total RNA extracted from wild-type, Mxi1.22, and Mxi1.14 U87 cell lines. RNAs were size-fractionated on a 1% agarose gel, blotted on a nylon membrane, hybridized with 32P-labelled cyclin B1, cyclin A and MXI1 cDNAs and assessed by autoradiography. Exposure was for 3 h (after overnight exposure the endogenous MXI1 mRNA was visible). To normalize RNA loading, the membrane was re-hybridized with the 32P-labelled cDNA of the housekeeping gene β-actin. The relative intensity of the bands was quantitated by densitometry.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2375210&req=5

fig5: Mxi1 inhibits expression of the cyclin B1 gene. Northern blot analysis was performed on total RNA extracted from wild-type, Mxi1.22, and Mxi1.14 U87 cell lines. RNAs were size-fractionated on a 1% agarose gel, blotted on a nylon membrane, hybridized with 32P-labelled cyclin B1, cyclin A and MXI1 cDNAs and assessed by autoradiography. Exposure was for 3 h (after overnight exposure the endogenous MXI1 mRNA was visible). To normalize RNA loading, the membrane was re-hybridized with the 32P-labelled cDNA of the housekeeping gene β-actin. The relative intensity of the bands was quantitated by densitometry.
Mentions: To dissect the molecular mechanism through which Mxi1 induces accumulation in the G2/M phase of the cell cycle, we investigated, in U87 proliferating cells, the expression of cyclin B1 gene and of cyclin A as control. The level of the two cyclin mRNAs was evaluated by Northern blot on RNA of U87.Mxi1.14, U87.Mxi1.22 and U87 wild-type cells. MXI1 and beta-actin transcripts were also analyzed on the same membrane. Figure 5Figure 5

Bottom Line: Interestingly, the expression of cyclin B1 was inhibited to about 60% in U87.Mxi1.14 cells.This inhibition occurs at the transcriptional level and depends, at least in part, on the E-box present on the cyclin B1 promoter.Consistent with this, the endogenous Mxi1 binds this E-box in vitro.

View Article: PubMed Central - PubMed

Affiliation: Istituto Regina Elena, Centro Ricerca Sperimentale, Laboratorio di Oncogenesi Molecolare, Via delle messi D'Oro 156, 00158 Rome, Italy.

Show MeSH
Related in: MedlinePlus