The detection of differentially expressed microRNAs from the serum of ovarian cancer patients using a novel real-time PCR platform

Gynecol Oncol. 2009 Jan;112(1):55-9. doi: 10.1016/j.ygyno.2008.08.036. Epub 2008 Oct 26.

Abstract

Objective: To determine the utility of serum miRNAs as biomarkers for epithelial ovarian cancer.

Methods: Twenty-eight patients with histologically confirmed epithelial ovarian cancer were identified from a tissue and serum bank. Serum was collected prior to definitive therapy. Fifteen unmatched, healthy controls were used for comparison. Serum was obtained from all patients. RNA was extracted using a derivation of the single step Trizol method. The RNA from 9 cancer specimens was compared to 4 normal specimens with real-time PCR using the TaqMan Array Human MicroRNA panel. Twenty-one miRNAs were differentially expressed between normal and patient serum. Real-time PCR for the 21 individual miRNAs was performed on the remaining 19 cancer specimens and 11 normal specimens.

Results: Eight miRNAs of the original twenty-one were identified that were significantly differentially expressed between cancer and normal specimens using the comparative C(t) method. MiRNAs-21, 92, 93, 126 and 29a were significantly over-expressed in the serum from cancer patients compared to controls (p<.01). MiRNAs-155, 127 and 99b were significantly under-expressed (p<.01). Additionally, miRs-21, 92 and 93 were over-expressed in 3 patients with normal pre-operative CA-125.

Conclusion: We demonstrate that the extraction of RNA and subsequent identification of miRNAs from the serum of individuals diagnosed with ovarian cancer is feasible. Real-time PCR-based microarray is a novel and practical means to performing high-throughput investigation of serum RNA samples. miRNAs-21, 92 and 93 are known oncogenes with therapeutic and biomarker potential.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Biomarkers, Tumor / blood*
  • Biomarkers, Tumor / genetics*
  • CA-125 Antigen / blood
  • Case-Control Studies
  • Female
  • Humans
  • MicroRNAs / blood*
  • Middle Aged
  • Neoplasm Staging
  • Oligonucleotide Array Sequence Analysis / methods
  • Ovarian Neoplasms / blood*
  • Ovarian Neoplasms / genetics*
  • Ovarian Neoplasms / pathology
  • Polymerase Chain Reaction / methods*
  • RNA, Neoplasm / blood*

Substances

  • Biomarkers, Tumor
  • CA-125 Antigen
  • MicroRNAs
  • RNA, Neoplasm