shRNA-mediated EMMPRIN silencing inhibits human leukemic monocyte lymphoma U937 cell proliferation and increases chemosensitivity to adriamycin.

Abstract

EMMPRIN is a widely distributed cell surface glycoprotein, which plays an important role in tumor progression and confers resistance to some chemotherapeutic drugs. Recent studies have shown that EMMPRIN overexpression indicates poor prognosis in acute myeloid leukemia (AML). However, little was known on the role of EMMPRIN in leukemia. Human leukemia cell line U937 was stably transfected with a EMMPRIN-targeted shRNA-containing vector to investigate the effect of EMMPRIN on cellular functions. EMMPRIN expression was monitored by qRT-PCR and Western blotting. Cell viability and proliferation were determined by trypan blue exclusion and BrdU labeling, respectively. Cell cycle and apoptosis were analyzed by flow cytometry. Cytotoxicity of chemotherapeutic agent adriamycin on cells was assessed by MTT assay. Knockdown of EMMPRIN gene significantly inhibited cell viability and decreased cell proliferation. Fluorescence-activated cell-sorting analysis revealed that the reduced EMMPRIN expression resulted in cell cycle arrest at G1 phase and induced apoptosis. Meanwhile, western blotting analysis showed that EMMPRIN knockdown was associated with downregulation of cell cycle- and apoptosis-related molecules including cyclin D1, cyclin E, as well as increase in cleavage of caspase-3 and PARP. This study also showed that silencing of EMMPRIN sensitized U937 cells to Adriamycin. EMMPRIN is involved in proliferation, growth, and chemosensitivity of human AML line U937, indicating that EMMPRIN may be a promising therapeutic target for AML.

• Authors: Hui Gao, Qixiao Jiang, Yantao Han, Jianjun Peng, Chunbo Wang
• Journal: Cell biochemistry and biophysics (Cell Biochem Biophys) Vol. 71 Issue 2 Pg. 827-35 (Mar 2015) ISSN: 1559-0283 [Electronic] United States
• PMID: 25260396 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Retracted Publication)

Chemical References

• Antineoplastic Agents
• RNA, Small Interfering
• Basigin
• Doxorubicin

Topics

• Antineoplastic Agents (pharmacology)
• Apoptosis (drug effects, genetics)
• Basigin (genetics)
• Cell Cycle Checkpoints (drug effects, genetics)
• Cell Proliferation (drug effects, genetics)
• Cell Survival (drug effects, genetics)
• Doxorubicin (pharmacology)
• G1 Phase (drug effects, genetics)
• Gene Expression Regulation, Neoplastic (drug effects, genetics)
• Gene Knockdown Techniques
• Gene Silencing
• Humans
• Leukemia (pathology)
• Lymphoma (pathology)
• RNA, Small Interfering (genetics)
• U937 Cells