Influence of recombinant lentiviral vector encoding miR-15a/16-1 in biological features of human nasopharyngeal carcinoma CNE-2Z cells

Cancer Biother Radiopharm. 2014 Dec;29(10):422-7. doi: 10.1089/cbr.2013.1596.

Abstract

MicroRNAs can function as tumor suppressor miRNAs. Bcl-2 is an antiapoptotic gene overexpressed in many tumors, including nasopharyngeal carcinoma (NPC). It is reported that microRNA-15a (miR-15a) and microRNA-16-1 (miR-16-1) could act as bcl-2 inhibitors. To investigate their effects on NPC, the authors used recombinant lentiviral vector to upregulate the expression of miR-15a/16-1 in NPC CNE-2Z cells. The authors divided cells into the control group, transfection group, radiotherapy group, and transfection-radiotherapy group. In this experiment, reverse transcription-quantitative polymerase chain reaction was used to detect the expression of miR-15a/16-1 and bcl-2 mRNA. Cell proliferation was analyzed by MTT assay. Flow cytometry was used to measure cell apoptosis. Radiosensitivity was measured using colony-forming experiment. The protein expression of bcl-2 was measured by western blot, the activation levels of caspase were detected by a spectrophotometric method. After transfection, cell proliferation was inhibited, while the apoptosis rate and radiosensitivity were increased. In addition, the activation of caspase-2 and caspase-3 was aggrandized correspondingly. Although the expression levels of bcl-2 mRNA in each group had no difference, the protein expression of bcl-2 was downregulated. These results suggested that miR-15a/16-1 could inhibit cell proliferation and increase the apoptosis and radiosensitivity of CNE-2 cells, by regulating the bcl-2 gene at post-transcriptional level and by increasing the activation of caspase-2 and caspase-3.

Keywords: apoptosis; bcl-2; microRNAs; nasopharyngeal carcinoma; radiosensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / genetics
  • Carcinoma
  • Caspase 2 / genetics
  • Caspase 3 / genetics
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • Cysteine Endopeptidases / genetics
  • Genetic Vectors / genetics*
  • Humans
  • Lentivirus / genetics*
  • MicroRNAs / genetics*
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms / genetics*
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • RNA, Messenger / genetics
  • Radiation Tolerance / genetics
  • Transfection / methods

Substances

  • MIRN15 microRNA, human
  • MIRN16 microRNA, human
  • MicroRNAs
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • CASP2 protein, human
  • Caspase 2
  • Caspase 3
  • Cysteine Endopeptidases