Detection of CCN1 and CCN5 mRNA in Human Cancer Samples Using a Modified In Situ Hybridization Technique

Methods Mol Biol. 2017:1489:495-504. doi: 10.1007/978-1-4939-6430-7_41.

Abstract

In situ hybridization is an ideal tool for the detection and localization of mRNA expression of specific gene(s) in tissue sections and cell lines for prognosis, predictive markers, and highlighted potential therapeutic targets. Given the importance of CCN1 and CCN5 in breast and pancreatic cancer progression, these two secretory proteins could be novel therapeutic targets. Thus, evaluating the distribution of mRNA of these targets using in situ hybridization could be important preclinical tools. This chapter describes a detailed in situ hybridization technique for the detection of CCN1 and CCN5 in formalin-fixed, paraffin-embedded patient samples of breast and pancreatic cancers.

Keywords: CCN1; CCN5; In situ hybridization; Probe preparation; RNA isolation; Reverse-transcriptase polymerase chain reaction.

MeSH terms

  • CCN Intercellular Signaling Proteins / genetics*
  • Cell Line, Tumor
  • Cysteine-Rich Protein 61 / genetics*
  • Humans
  • In Situ Hybridization* / methods
  • RNA Probes
  • RNA, Messenger / genetics*
  • Repressor Proteins / genetics*

Substances

  • CCN Intercellular Signaling Proteins
  • CCN1 protein, human
  • CCN5 protein, human
  • Cysteine-Rich Protein 61
  • RNA Probes
  • RNA, Messenger
  • Repressor Proteins