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Absolute quantification of cell-free microRNAs in cancer patients.

Ferracin M, Lupini L, Salamon I, Saccenti E, Zanzi MV, Rocchi A, Da Ros L, Zagatti B, Musa G, Bassi C, Mangolini A, Cavallesco G, Frassoldati A, Volpato S, Carcoforo P, Hollingsworth AB, Negrini M - Oncotarget (2015)

Bottom Line: Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results.In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls.The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy.

ABSTRACT
The hypothesis to use microRNAs (miRNAs) circulating in the blood as cancer biomarkers was formulated some years ago based on promising initial results. After some exciting discoveries, however, it became evident that the accurate quantification of cell-free miRNAs was more challenging than expected. Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results. In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls. Then, we assessed the absolute level of nine different miRNAs (miR-320a, miR-21-5p, miR-378a-3p, miR-181a-5p, miR-3156-5p, miR-2110, miR-125a-5p, miR-425-5p, miR-766-3p) in 207 samples from healthy controls and cancer patients using droplet digital PCR (ddPCR) technology. We identified miRNAs specifically modulated in one or more cancer types, according to tissue source. The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively. This study finally powers the use of cell-free miRNAs as cancer biomarkers and propose miR-181a-5p as a diagnostic breast cancer biomarker.

No MeSH data available.


Related in: MedlinePlus

Correlation of miRNA levels in matched plasma-serumConcentration of miR-21-5p A. and miR-181a-5p B. in paired serum and plasma from the same subject. miR-21-5p levels are inversely correlated (Pearson r = −0.32, p = 0.055); miR-181a-5p levels are positively correlated (Pearson r = 0.31, p = 0.023) in the two tissues.
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Figure 3: Correlation of miRNA levels in matched plasma-serumConcentration of miR-21-5p A. and miR-181a-5p B. in paired serum and plasma from the same subject. miR-21-5p levels are inversely correlated (Pearson r = −0.32, p = 0.055); miR-181a-5p levels are positively correlated (Pearson r = 0.31, p = 0.023) in the two tissues.

Mentions: We assessed miR-21-5p (Figure 3A) and miR-181a-5p (Figure 3B) levels in randomly selected, matched, plasma and sera from the same patient. Considering that plasma was prepared using a one-step centrifugation at 1000 x g, we found substantially different levels of the same miRNA in the two tissues. Pearson correlation index was −0.32 (p = 0.055) for miR-21-5p and 0.3 (p = 0.024) for miR-181a-5p. These results suggested that the correlation of miRNA levels in matched plasma and serum can vary for each miRNA species and rely strongly on tissue preparation.


Absolute quantification of cell-free microRNAs in cancer patients.

Ferracin M, Lupini L, Salamon I, Saccenti E, Zanzi MV, Rocchi A, Da Ros L, Zagatti B, Musa G, Bassi C, Mangolini A, Cavallesco G, Frassoldati A, Volpato S, Carcoforo P, Hollingsworth AB, Negrini M - Oncotarget (2015)

Correlation of miRNA levels in matched plasma-serumConcentration of miR-21-5p A. and miR-181a-5p B. in paired serum and plasma from the same subject. miR-21-5p levels are inversely correlated (Pearson r = −0.32, p = 0.055); miR-181a-5p levels are positively correlated (Pearson r = 0.31, p = 0.023) in the two tissues.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4546486&req=5

Figure 3: Correlation of miRNA levels in matched plasma-serumConcentration of miR-21-5p A. and miR-181a-5p B. in paired serum and plasma from the same subject. miR-21-5p levels are inversely correlated (Pearson r = −0.32, p = 0.055); miR-181a-5p levels are positively correlated (Pearson r = 0.31, p = 0.023) in the two tissues.
Mentions: We assessed miR-21-5p (Figure 3A) and miR-181a-5p (Figure 3B) levels in randomly selected, matched, plasma and sera from the same patient. Considering that plasma was prepared using a one-step centrifugation at 1000 x g, we found substantially different levels of the same miRNA in the two tissues. Pearson correlation index was −0.32 (p = 0.055) for miR-21-5p and 0.3 (p = 0.024) for miR-181a-5p. These results suggested that the correlation of miRNA levels in matched plasma and serum can vary for each miRNA species and rely strongly on tissue preparation.

Bottom Line: Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results.In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls.The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Morphology, Surgery and Experimental Medicine, University of Ferrara, Ferrara, Italy.

ABSTRACT
The hypothesis to use microRNAs (miRNAs) circulating in the blood as cancer biomarkers was formulated some years ago based on promising initial results. After some exciting discoveries, however, it became evident that the accurate quantification of cell-free miRNAs was more challenging than expected. Difficulties were linked to the strong impact that many, if not all, pre- and post- analytical variables have on the final results. In this study, we used currently available high-throughput technologies to identify miRNAs present in plasma and serum of patients with breast, colorectal, lung, thyroid and melanoma tumors, and healthy controls. Then, we assessed the absolute level of nine different miRNAs (miR-320a, miR-21-5p, miR-378a-3p, miR-181a-5p, miR-3156-5p, miR-2110, miR-125a-5p, miR-425-5p, miR-766-3p) in 207 samples from healthy controls and cancer patients using droplet digital PCR (ddPCR) technology. We identified miRNAs specifically modulated in one or more cancer types, according to tissue source. The significant reduction of miR-181a-5p levels in breast cancer patients serum was further validated using two independent cohorts, one from Italy (n = 70) and one from US (n = 90), with AUC 0.66 and 0.73 respectively. This study finally powers the use of cell-free miRNAs as cancer biomarkers and propose miR-181a-5p as a diagnostic breast cancer biomarker.

No MeSH data available.


Related in: MedlinePlus