Effects of CDC7 gene silencing and Rituximab on apoptosis in diffuse large B cell lymphoma cells

J Cancer Res Clin Oncol. 2012 Dec;138(12):2027-34. doi: 10.1007/s00432-012-1283-2. Epub 2012 Jul 18.

Abstract

Background: The cell division cycle 7 (CDC7) is a serine-threonine kinase, which is required for DNA replication and is high expressed in diffuse large B cell lymphoma (DLBCL).

Methods: In this study, we targeted CDC7 in human DLBCL-ABC subtype cells (ly3) and examined the subsequent alterations in cellular apoptosis. The expression of CDC7 was silenced with small interfering RNA (siRNA)-expressing plasmid. CDC7 gene silencing cells were then incubated with or without rituximab for 24 h. Following treatment, Annexin V/propidium iodide staining followed by flow cytometry was used to examine cellular apoptosis. Furthermore, the expression of caspase 3, Bax, and Bcl-2 protein was analyzed by Western blotting. The expression of Bax and Bcl-2 mRNA was analyzed by quantitative real-time PCR.

Results: Compared to non-treated or control siRNA-transfected cells, significantly higher levels of apoptosis were detected in siCDC7-transfected cells and rituximab-treated cells (P < 0.05), which was further enhanced in CDC7-targeted cells with rituximab treatment (P < 0.05). The pro-apoptotic effects were accompanied with up-regulation of caspase 3 and Bax, meanwhile down-regulation of Bcl-2.

Conclusion: Combined treatments using rituximab and CDC7 gene silencing significantly increases apoptosis in ly3 cells and plays synergistic effect. CDC7 is a novel therapeutic target in DLBCL patients. CDC7 inhibitors combined with rituximab will be the new therapy for the ABC-DLBCL patients.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal, Murine-Derived / pharmacology*
  • Antineoplastic Agents / pharmacology*
  • Apoptosis / genetics*
  • Caspase 3 / genetics
  • Caspase 3 / metabolism
  • Cell Cycle Proteins / genetics*
  • Cell Cycle Proteins / metabolism
  • Cell Line, Tumor
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Gene Silencing*
  • Humans
  • Lymphoma, Large B-Cell, Diffuse / drug therapy*
  • Lymphoma, Large B-Cell, Diffuse / genetics*
  • Lymphoma, Large B-Cell, Diffuse / metabolism
  • Phosphorylation
  • Protein Serine-Threonine Kinases / genetics*
  • Protein Serine-Threonine Kinases / metabolism
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Rituximab
  • Up-Regulation / drug effects
  • Up-Regulation / genetics
  • bcl-2-Associated X Protein / genetics
  • bcl-2-Associated X Protein / metabolism

Substances

  • Antibodies, Monoclonal, Murine-Derived
  • Antineoplastic Agents
  • Cell Cycle Proteins
  • RNA, Messenger
  • bcl-2-Associated X Protein
  • Rituximab
  • CDC7 protein, human
  • Protein Serine-Threonine Kinases
  • Caspase 3