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miRNAs associated with chemo-sensitivity in cell lines and in advanced bladder cancer.

Nordentoft I, Birkenkamp-Demtroder K, Agerbæk M, Theodorescu D, Ostenfeld MS, Hartmann A, Borre M, Ørntoft TF, Dyrskjøt L - BMC Med Genomics (2012)

Bottom Line: Three miRNAs were associated with both response and survival (886-3p, 923, 944).By changing the cellular level of the response-identified miRNAs in eight bladder cell lines with different cisplatin sensitivity we found that down-regulation of miR-27a, miR296-5p and miR-642 generally reduced the cell viability, whereas up-regulation of miR-138 and miR-886-3p reduced the viability of more than half of the cell lines.Decreasing miR-138 increased the cisplatin sensitivity in half of the cell lines and increasing miR-27a and miR-642 generally increased cisplatin sensitivity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark. iver.nordentoft@ki.au.dk

ABSTRACT

Background: MicroRNA is a naturally occurring class of non-coding RNA molecules that mediate posttranscriptional gene regulation and are strongly implicated in cellular processes such as cell proliferation, carcinogenesis, cell survival and apoptosis. Consequently there is increasing focus on miRNA expression as prognostic factors for outcome and chemotherapy response. Only approximately 50% of patients with bladder cancer respond to chemotherapy. Therefore, predictive markers, such as miRNAs, that can identify subgroups of patients who will benefit from chemotherapy will have great value for treatment guidance.

Methods: We profiled the expression of 671 miRNAs in formalin fixed paraffin embedded tumors from patients with advanced bladder cancer treated with cisplatin based chemotherapy. We delineated differentially expressed miRNAs in tumors from patients with complete response vs. patients with progressive disease and in tumors form patients with short and long overall survival time. Furthermore, we studied the effect of up- and down regulation of key miRNAs on the cisplatin sensitivity in eight bladder cancer cell lines with different sensitivities to cisplatin.

Results: miRNA expression profiling identified 15 miRNAs that correlated with response to chemotherapy and 5 miRNAs that correlated with survival time. Three miRNAs were associated with both response and survival (886-3p, 923, 944). By changing the cellular level of the response-identified miRNAs in eight bladder cell lines with different cisplatin sensitivity we found that down-regulation of miR-27a, miR296-5p and miR-642 generally reduced the cell viability, whereas up-regulation of miR-138 and miR-886-3p reduced the viability of more than half of the cell lines. Decreasing miR-138 increased the cisplatin sensitivity in half of the cell lines and increasing miR-27a and miR-642 generally increased cisplatin sensitivity.

Conclusions: MiRNAs seem to be involved in cisplatin based chemo response and may form a new target for therapy and serve as biomarkers for treatment response.

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Single agent dose-dependent cytotoxicity induced by cisplatin. Human bladder cancer cells UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A were seeded in 96 well plates and treatment with the indicated cisplatin concentration started 48 h after seeding and continued for 48 h. Cell viability was assessed with MTT-assay. Panel A: cell viability cisplatin dose response curve. Panel B: Cisplatin concentration killing 50% of the cells after 48 h relative to non-treated cells (GI50, Growth inhibition 50%). GI50 was estimated based on two independent experiments. Each drug concentration was tested in six individual wells.
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Figure 2: Single agent dose-dependent cytotoxicity induced by cisplatin. Human bladder cancer cells UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A were seeded in 96 well plates and treatment with the indicated cisplatin concentration started 48 h after seeding and continued for 48 h. Cell viability was assessed with MTT-assay. Panel A: cell viability cisplatin dose response curve. Panel B: Cisplatin concentration killing 50% of the cells after 48 h relative to non-treated cells (GI50, Growth inhibition 50%). GI50 was estimated based on two independent experiments. Each drug concentration was tested in six individual wells.

Mentions: In order to investigate the ability of the differentially expressed miRNAs from patient response cohort to predict the sensitivity to cisplatin based chemotherapy we exposed a range of bladder cancer cell lines (UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A) to cisplatin. These cell lines have previously been documented to exhibit differing sensitivity to cisplatin. Above the cell are listed from high to low cisplatin sensitivity. The concentration of cisplatin required to inhibit the cell growth by 50% (GI50) during 48 h incubation in our laboratory was calculated from dose response curves (Figure 2). The studies were conducted with equal cell seeding density to avoid any confluences-related difference in effect observed on cell viability. Next we measured the expression of miRNA-27a, miRNA-138, miRNA-193a-5p, miRNA-296-5p, miRNA-410, miRNA-492, miRNA-642, miRNA-886-3p, miRNA-923 and miRNA-944 using qRT-PCR in the 8 cell lines (Figure 3). We observed no association between cisplatin GI50 and miRNA expression in the cell lines.


miRNAs associated with chemo-sensitivity in cell lines and in advanced bladder cancer.

Nordentoft I, Birkenkamp-Demtroder K, Agerbæk M, Theodorescu D, Ostenfeld MS, Hartmann A, Borre M, Ørntoft TF, Dyrskjøt L - BMC Med Genomics (2012)

Single agent dose-dependent cytotoxicity induced by cisplatin. Human bladder cancer cells UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A were seeded in 96 well plates and treatment with the indicated cisplatin concentration started 48 h after seeding and continued for 48 h. Cell viability was assessed with MTT-assay. Panel A: cell viability cisplatin dose response curve. Panel B: Cisplatin concentration killing 50% of the cells after 48 h relative to non-treated cells (GI50, Growth inhibition 50%). GI50 was estimated based on two independent experiments. Each drug concentration was tested in six individual wells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3473298&req=5

Figure 2: Single agent dose-dependent cytotoxicity induced by cisplatin. Human bladder cancer cells UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A were seeded in 96 well plates and treatment with the indicated cisplatin concentration started 48 h after seeding and continued for 48 h. Cell viability was assessed with MTT-assay. Panel A: cell viability cisplatin dose response curve. Panel B: Cisplatin concentration killing 50% of the cells after 48 h relative to non-treated cells (GI50, Growth inhibition 50%). GI50 was estimated based on two independent experiments. Each drug concentration was tested in six individual wells.
Mentions: In order to investigate the ability of the differentially expressed miRNAs from patient response cohort to predict the sensitivity to cisplatin based chemotherapy we exposed a range of bladder cancer cell lines (UMUC9, UMUC14, SLT4, 253JBV, RT4, CLR2169, HT1197, 575A) to cisplatin. These cell lines have previously been documented to exhibit differing sensitivity to cisplatin. Above the cell are listed from high to low cisplatin sensitivity. The concentration of cisplatin required to inhibit the cell growth by 50% (GI50) during 48 h incubation in our laboratory was calculated from dose response curves (Figure 2). The studies were conducted with equal cell seeding density to avoid any confluences-related difference in effect observed on cell viability. Next we measured the expression of miRNA-27a, miRNA-138, miRNA-193a-5p, miRNA-296-5p, miRNA-410, miRNA-492, miRNA-642, miRNA-886-3p, miRNA-923 and miRNA-944 using qRT-PCR in the 8 cell lines (Figure 3). We observed no association between cisplatin GI50 and miRNA expression in the cell lines.

Bottom Line: Three miRNAs were associated with both response and survival (886-3p, 923, 944).By changing the cellular level of the response-identified miRNAs in eight bladder cell lines with different cisplatin sensitivity we found that down-regulation of miR-27a, miR296-5p and miR-642 generally reduced the cell viability, whereas up-regulation of miR-138 and miR-886-3p reduced the viability of more than half of the cell lines.Decreasing miR-138 increased the cisplatin sensitivity in half of the cell lines and increasing miR-27a and miR-642 generally increased cisplatin sensitivity.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark. iver.nordentoft@ki.au.dk

ABSTRACT

Background: MicroRNA is a naturally occurring class of non-coding RNA molecules that mediate posttranscriptional gene regulation and are strongly implicated in cellular processes such as cell proliferation, carcinogenesis, cell survival and apoptosis. Consequently there is increasing focus on miRNA expression as prognostic factors for outcome and chemotherapy response. Only approximately 50% of patients with bladder cancer respond to chemotherapy. Therefore, predictive markers, such as miRNAs, that can identify subgroups of patients who will benefit from chemotherapy will have great value for treatment guidance.

Methods: We profiled the expression of 671 miRNAs in formalin fixed paraffin embedded tumors from patients with advanced bladder cancer treated with cisplatin based chemotherapy. We delineated differentially expressed miRNAs in tumors from patients with complete response vs. patients with progressive disease and in tumors form patients with short and long overall survival time. Furthermore, we studied the effect of up- and down regulation of key miRNAs on the cisplatin sensitivity in eight bladder cancer cell lines with different sensitivities to cisplatin.

Results: miRNA expression profiling identified 15 miRNAs that correlated with response to chemotherapy and 5 miRNAs that correlated with survival time. Three miRNAs were associated with both response and survival (886-3p, 923, 944). By changing the cellular level of the response-identified miRNAs in eight bladder cell lines with different cisplatin sensitivity we found that down-regulation of miR-27a, miR296-5p and miR-642 generally reduced the cell viability, whereas up-regulation of miR-138 and miR-886-3p reduced the viability of more than half of the cell lines. Decreasing miR-138 increased the cisplatin sensitivity in half of the cell lines and increasing miR-27a and miR-642 generally increased cisplatin sensitivity.

Conclusions: MiRNAs seem to be involved in cisplatin based chemo response and may form a new target for therapy and serve as biomarkers for treatment response.

Show MeSH
Related in: MedlinePlus