An rGONS-based biosensor for simultaneous imaging of p53 and p21 mRNA in living cells

Talanta. 2019 Nov 1:204:20-28. doi: 10.1016/j.talanta.2019.05.087. Epub 2019 May 24.

Abstract

Hepatocarcinoma is the second leading cause of cancer-related death worldwide accompanied by the aberrant expression of many genes. Among of them, p53 and p21 genes played a vital role in the development of liver cancer. Thus, monitoring mRNA levels of the two markers is urgently needed for early diagnosis and understanding the molecular mechanism of hepatocarcinoma development. Herein, a functional nanosystem for in situ and real-time monitoring levels of p53 and p21 mRNA was constructed using reduced graphene oxide nanosheet assisted fluorescence probes. Comparing with traditional methods, the new method indicated high sensitivity and outstanding selectivity towards p53 and p21 mRNA assay in vitro. Moreover, the functional nanosystem was successfully used for monitoring dynamic change of mRNAs in HepG2 cells caused by cisplatin treatment, the reliability of which was confirmed by RT-PCR. In summary, this strategy provides a promising tool to reveal p53 and p21 mRNA regulatory process in cancer cells, which is practicable for drug screening and therapy evaluation on clinic.

Keywords: Cell imaging; Nanosystem; mRNA; p21; p53; rGONS.

MeSH terms

  • Adsorption
  • Antineoplastic Agents / pharmacology
  • Biosensing Techniques / methods
  • Cell Line, Tumor
  • Cisplatin / pharmacology
  • Cyclin-Dependent Kinase Inhibitor p21 / genetics
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism*
  • DNA / chemistry
  • DNA / genetics
  • DNA Probes / chemistry
  • DNA Probes / genetics
  • Fluorescent Dyes / chemistry
  • Graphite / chemistry*
  • Humans
  • Limit of Detection
  • Microscopy, Confocal / methods
  • Microscopy, Fluorescence / methods
  • Nanostructures / chemistry*
  • Nucleic Acid Hybridization
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Tumor Suppressor Protein p53 / genetics
  • Tumor Suppressor Protein p53 / metabolism*

Substances

  • Antineoplastic Agents
  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • DNA Probes
  • Fluorescent Dyes
  • RNA, Messenger
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • graphene oxide
  • Graphite
  • DNA
  • Cisplatin