Antisense epidermal growth factor receptor RNA transfection in human malignant glioma cells leads to inhibition of proliferation and induction of differentiation.
The epidermal growth factor receptor (EGFR) is a protooncogene that is frequently observed with alterations in late stage gliomas, suggesting an important role of this gene in glial tumorigenesis and progression. In this study we evaluated an antisense EGFR approach as an alternative therapeutic modality for glioblastomas. We transfected U-87MG cells with an antisense EGFR construct and obtained several clones stably expressing lower or undetectable levels of EGFR protein. These clones were found to have impaired proliferation as well as a reduced transforming potential to grow in soft agarose. The number of cells positive for the cell cycle-specific nuclear antigen Ki-67 was also significantly decreased (P < 0.05) in antisense EGFR-transfected clones compared with parental or empty vector-transfected cells. Flow cytometric analysis revealed that the proportion of cells in G0/G1 phases of the cell cycle in the antisense clones increased by up to 31% compared with control cells, whereas the proportion of cells in S phase decreased by up to 58%. In addition, the antisense EGFR-transfected cells showed higher expression of glial fibrillary acidic protein and a more differentiated form, with smaller cell bodies possessing fine tapering cell processes. These results suggest that EGFR plays a major role in modulating cell growth and differentiation in glioblastoma cells. Our experimental model of antisense EGFR provides a basis for future development of antisense EGFR oligodeoxynucleotides in treatment of glioblastomas.
Tian
X X
XX
Department of Anatomical & Cellular Pathology, Prince of Wales Hospital, Chinese University of Hong Kong, Shatin, China.
Lam
P Y
PY
Chen
J
J
Pang
J C
JC
To
S S
SS
Di-Tomaso
E
E
Ng
H K
HK
D000911:Antibodies, Monoclonal; D016375:Antisense Elements (Genetics) / Q000494:pharmacology*; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D066246:ErbB Receptors / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005909:Glioblastoma / Q000628:therapy*; D006801:Humans; D019394:Ki-67 Antigen / Q000032:analysis; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000166:cytology
D000911:Antibodies, Monoclonal; D016375:Antisense Elements (Genetics); D005904:Glial Fibrillary Acidic Protein; D019394:Ki-67 Antigen; D066246:ErbB Receptors