Ann Surg Treat Res.  2014 Dec;87(6):290-297. 10.4174/astr.2014.87.6.290.

MicroRNA expression profiling of diagnostic needle aspirates from surgical pancreatic cancer specimens

• Hong TH
• Park IY

• KMID: 2167084
• DOI: http://doi.org/10.4174/astr.2014.87.6.290

Abstract

PURPOSE
MicroRNAs (miRNAs) have been widely investigated as potential biomarkers for several malignancies. To establish the feasibility of miRNA expression profiling of small biopsy samples of pancreatic cancers, we assessed expression profiles in freshly collected aspirates obtained immediately after surgical resection of the pancreas.
METHODS
We used separate fine needles (20-23 gauge) to aspirate the pancreatic cancer and adjacent normal pancreatic tissue. miRNAs that were differentially expressed in pancreatic cancers and matched paraneoplastic normal pancreatic tissues were identified using an miRNA microarray.
RESULTS
We identified 158 aberrantly expressed miRNAs in pancreatic cancers; 51 were overexpressed and 107 underexpressed compared with normal pancreatic tissue. To confirm the microarray findings, quantitative RT-PCR was performed on individual samples. We chose eight miRNAs for further analysis; of which five were overexpressed (miR-21, miR-27a, miR-146a, miR-200a, and miR-196a) and three underexpressed (miR-217, miR-20a, and miR-96) in pancreatic cancer samples compared to benign pancreatic tissue. Expression of miR-21, miR-27a, miR-146a, miR-200a, and miR-196a was significantly increased in cancer fine-needle aspirates relative to matched controls in all samples. Expression of miR-217, miR-20a, and miR-96 was significantly downregulated in almost all pancreatic cancer tissues.
CONCLUSION
We demonstrate the feasibility of performing miRNA profiling on very small specimens obtained using fine-needle aspiration of pancreatic cancers.

Keyword

MicroRNAs; Pancreatic neoplasms; Biopsy; Fine-needle

MeSH Terms

Biomarkers
Biopsy
Biopsy, Fine-Needle
MicroRNAs*
Needles*
Pancreas
Pancreatic Neoplasms*
Biological Markers
MicroRNAs

Figure

• 

  Fig. 1 A heat map and hierarchical clustering showing the overall expression patterns of microRNAs. Red represents an expression level higher than that of the normal control, and green lower expression.

• 

  Fig. 2 Comparative expression analysis of miR-21 (A), miR-27a (B), miR-146a (C), miR-200a (D), miR-196a (E), miR-217 (F), miR-20a (G), and miR-96 (H) levels in 11 fine-needle aspiration specimens from cancer relative to matched normal control tissue, as estimated by quantitative RT-PCR. The expression levels in control paraneoplastic normal pancreatic tissues were set to 1.0 in terms of log2 values.

• 

  Fig. 3 Box plot showing the overall expression levels of eight species of microRNAs (miRNAs) as revealed by quantitative RT-PCR (qRT-PCR) in 11 fine-needle aspiration specimens from cancer compared to control tissue. The expression levels in control paraneoplastic normal pancreatic tissues were set to 1.0 in terms of log2 values.

• 

  Fig. 4 Box plot showing the expression levels of eight species of microRNAs (miRNAs), assessed using quantitative RT-PCR (qRT-PCR), in 19 cryopreserved sections of pancreatic cancer tissue compared with 11 control tissues. The expression levels in control paraneoplastic normal pancreatic tissues were set to 1.0 in terms of log2 values.

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