Knockdown of cyclin D1 inhibits proliferation, induces apoptosis, and attenuates the invasive capacity of human glioblastoma cells.

Abstract

Elevated cyclin D1 (CCND1) in human glioblastoma correlates with poor clinical prognosis. In this study, the human glioblastoma cell lines SHG-44 and U251 were stably transfected with short hairpin RNA (shRNA) targeting cyclin D1 or with ectogenic cyclin D1 by lentivirus-mediated transfection. Glioblastoma cells overexpressing or underexpressing cyclin D1 were then examined by in vitro growth assays, apoptosis assays, cell cycle analysis, and invasion assays. Cyclin D1 knockdown in SHG-44 cells inhibited cell proliferation, induced apoptosis, and attenuated migration across Matrigel, a model of invasive capacity. Western blot analysis and quantitative reverse-transcription polymerase chain reaction (RT-PCR) revealed that cells underexpressing CCND1 exhibited decreased multidrug resistance protein 1 (MDR1) and B-cell lymphoma-2 (Bcl-2) expression, but enhanced apoptosis effector caspase-3 expression. In contrast, cyclin D1 overexpression promoted cell proliferation, attenuated apoptosis, and enhanced invasive capacity. Furthermore, cyclin D1 overexpression was associated with increased expression of MDR1 and Bcl-2, and decreased caspase-3 expression. Results using the U251 cell line confirmed the effects of CCND1-targeted shRNA and lentivirus-mediated overexpression on proliferation and apoptosis of glioblastoma cells. Overexpression of cyclin D1 enhanced the proliferation and invasive potential of human glioblastoma cells, while reducing apoptosis. The ability to suppress the malignant phenotype by downregulating cyclin D1 expression may provide a new gene therapy approach for patients with malignant glioma.

• Authors: Junyu Wang, Qi Wang, Yong Cui, Zhen Yang Liu, Wei Zhao, Chun Lin Wang, Yan Dong, Lijun Hou, Guohan Hu, Chun Luo, Juxiang Chen, Yicheng Lu
• Journal: Journal of neuro-oncology (J Neurooncol) Vol. 106 Issue 3 Pg. 473-84 (Feb 2012) ISSN: 1573-7373 [Electronic] United States
• PMID: 21912938 (Publication Type: Journal Article)

Chemical References

• ATP Binding Cassette Transporter, Subfamily B, Member 1
• Annexin A5
• CCND1 protein, human
• Proto-Oncogene Proteins c-bcl-2
• RNA, Small Interfering
• Cyclin D1
• Green Fluorescent Proteins
• Caspase 3
• Matrix Metalloproteinase 2
• Matrix Metalloproteinase 9

Topics

• ATP Binding Cassette Transporter, Subfamily B, Member 1 (genetics, metabolism)
• Analysis of Variance
• Annexin A5 (metabolism)
• Apoptosis (drug effects, physiology)
• Caspase 3 (metabolism)
• Cell Cycle (drug effects)
• Cell Line, Transformed
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Cyclin D1 (metabolism)
• Flow Cytometry
• Gene Expression Regulation, Neoplastic (drug effects, physiology)
• Glioblastoma (pathology)
• Green Fluorescent Proteins (genetics, metabolism)
• Humans
• Matrix Metalloproteinase 2 (metabolism)
• Matrix Metalloproteinase 9 (metabolism)
• Neoplasm Invasiveness (genetics, prevention & control)
• Proto-Oncogene Proteins c-bcl-2 (genetics, metabolism)
• RNA, Small Interfering (pharmacology)
• Time Factors
• Transfection