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SV40 associated miRNAs are not detectable in mesotheliomas.

Gee GV, Stanifer ML, Christensen BC, Atwood WJ, Ugolini D, Bonassi S, Resnick MB, Nelson HH, Marsit CJ, Kelsey KT - Br. J. Cancer (2010)

Bottom Line: The Institute of Medicine has recommended the development of more sensitive and specific tests to resolve this controversy.Using this sensitive and specific detection method, we were unable to identify SV40 miRNA expression in human malignant pleural mesothelioma (MM) samples.Our work indicates that SV40 miRNAs are not likely to contribute to mesothelioma tumourogenesis, but highlights the value of this approach when compared with the relatively unspecific current testing methods.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Brown University, Box G-E-5, Providence, RI 02912, USA.

ABSTRACT

Background: Simian virus-40 (SV40) is a DNA tumour virus that was introduced into the human population with contaminated poliovirus vaccine, and its role in mesothelioma is widely debated. PCR based testing has been called into question, as false positives can be because of cross-reactivity with related viruses, or to laboratory contamination. The Institute of Medicine has recommended the development of more sensitive and specific tests to resolve this controversy.

Methods: We have characterized highly sensitive RT-PCR based assays that are specific for SV40-encoded microRNAs (miRNAs), as an alternative to current testing methods.

Results: Using this sensitive and specific detection method, we were unable to identify SV40 miRNA expression in human malignant pleural mesothelioma (MM) samples.

Conclusion: Our work indicates that SV40 miRNAs are not likely to contribute to mesothelioma tumourogenesis, but highlights the value of this approach when compared with the relatively unspecific current testing methods.

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Related in: MedlinePlus

Quantitative RT–PCR of SV40 microRNA. Quantitative RT–PCR was performed on 94 human malignant mesothelioma biopsies, 28 nonmalignant biopsies and 20 lung adenocarcinomas. Quantitative PCR was performed to measure the levels of miRNA relative to the human control small RNA, RNU48. The relative quantification of miRNA was performed according to the 2−ΔΔCt method and measurements were made relative to RNA isolated from HEK cells 72 h after SV40 infection.
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fig4: Quantitative RT–PCR of SV40 microRNA. Quantitative RT–PCR was performed on 94 human malignant mesothelioma biopsies, 28 nonmalignant biopsies and 20 lung adenocarcinomas. Quantitative PCR was performed to measure the levels of miRNA relative to the human control small RNA, RNU48. The relative quantification of miRNA was performed according to the 2−ΔΔCt method and measurements were made relative to RNA isolated from HEK cells 72 h after SV40 infection.

Mentions: We used the minimal 5′ and 3′ assays to detect the expression of SV40-encoded miRNAs in 94 human malignant mesothelioma tumour biopsies as well as in 20 lung adenocarcinoma cancerous and 28 non-malignant control biopsies (Figure 4). Despite the high sensitivity of these assays, we were unable to find any evidence of SV40 miRNA expression in human tissues. To ensure that the lack of SV40 miRNA expression in mesotheliomas was not because of technical problems isolating miRNA from these tissue samples, or to a general miRNA processing defect of this tumour type, we used quantitative PCR to detect three separate host-encoded miRNAs in mesotheliomas compared with non-malignant lung tissue (Supplementary Figure 1).


SV40 associated miRNAs are not detectable in mesotheliomas.

Gee GV, Stanifer ML, Christensen BC, Atwood WJ, Ugolini D, Bonassi S, Resnick MB, Nelson HH, Marsit CJ, Kelsey KT - Br. J. Cancer (2010)

Quantitative RT–PCR of SV40 microRNA. Quantitative RT–PCR was performed on 94 human malignant mesothelioma biopsies, 28 nonmalignant biopsies and 20 lung adenocarcinomas. Quantitative PCR was performed to measure the levels of miRNA relative to the human control small RNA, RNU48. The relative quantification of miRNA was performed according to the 2−ΔΔCt method and measurements were made relative to RNA isolated from HEK cells 72 h after SV40 infection.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2966630&req=5

fig4: Quantitative RT–PCR of SV40 microRNA. Quantitative RT–PCR was performed on 94 human malignant mesothelioma biopsies, 28 nonmalignant biopsies and 20 lung adenocarcinomas. Quantitative PCR was performed to measure the levels of miRNA relative to the human control small RNA, RNU48. The relative quantification of miRNA was performed according to the 2−ΔΔCt method and measurements were made relative to RNA isolated from HEK cells 72 h after SV40 infection.
Mentions: We used the minimal 5′ and 3′ assays to detect the expression of SV40-encoded miRNAs in 94 human malignant mesothelioma tumour biopsies as well as in 20 lung adenocarcinoma cancerous and 28 non-malignant control biopsies (Figure 4). Despite the high sensitivity of these assays, we were unable to find any evidence of SV40 miRNA expression in human tissues. To ensure that the lack of SV40 miRNA expression in mesotheliomas was not because of technical problems isolating miRNA from these tissue samples, or to a general miRNA processing defect of this tumour type, we used quantitative PCR to detect three separate host-encoded miRNAs in mesotheliomas compared with non-malignant lung tissue (Supplementary Figure 1).

Bottom Line: The Institute of Medicine has recommended the development of more sensitive and specific tests to resolve this controversy.Using this sensitive and specific detection method, we were unable to identify SV40 miRNA expression in human malignant pleural mesothelioma (MM) samples.Our work indicates that SV40 miRNAs are not likely to contribute to mesothelioma tumourogenesis, but highlights the value of this approach when compared with the relatively unspecific current testing methods.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Laboratory Medicine, Brown University, Box G-E-5, Providence, RI 02912, USA.

ABSTRACT

Background: Simian virus-40 (SV40) is a DNA tumour virus that was introduced into the human population with contaminated poliovirus vaccine, and its role in mesothelioma is widely debated. PCR based testing has been called into question, as false positives can be because of cross-reactivity with related viruses, or to laboratory contamination. The Institute of Medicine has recommended the development of more sensitive and specific tests to resolve this controversy.

Methods: We have characterized highly sensitive RT-PCR based assays that are specific for SV40-encoded microRNAs (miRNAs), as an alternative to current testing methods.

Results: Using this sensitive and specific detection method, we were unable to identify SV40 miRNA expression in human malignant pleural mesothelioma (MM) samples.

Conclusion: Our work indicates that SV40 miRNAs are not likely to contribute to mesothelioma tumourogenesis, but highlights the value of this approach when compared with the relatively unspecific current testing methods.

Show MeSH
Related in: MedlinePlus