A vector-based short hairpin RNA targeting Aurora A inhibits breast cancer growth.
Aurora A plays an essential role in centrosome maturation, separation and in the formation of the mitotic bipolar spindle. Overexpression or amplication of Aurora A gene has been detected in many cancer cell lines and various tumor tissues, including breast cancer, suggesting that Aurora A might be drug target for breast cancer treatment. In the current study, short hairpin RNA targeting Aurora A was cloned into pGenesil-2 plasmid vector and then transfected into MDA-MB-435S and ZR-75-30 human breast cancer cells using cationic liposome. Reduced expression of Aurora A was detected by RT-PCR and Western blot. The effect of pGenesil-2-shAURKA plasmid on tumor growth in MDA-MB-435S xenogenic implantation model was studied. pGenesil-2-shAURKA plasmid inhibited tumor growth significantly by systemantic administration. To further study the underlying mechanisms, cell apoptosis and proliferation were investigated by flow cytometric analysis, propidium iodide staining, TUNEL and Ki-67 immunostaining respectively. Increased apoptosis and reduced cell proliferation were detected in vitro and in vivo studies. In summary, our results suggested that specific knockdown of Aurora A expression by vector based shRNA may be a potential therapy for human breast cancer.
Wan
Yang
Y
State Key Laboratory of Biotherapy, West China Hospital and West China Medical School, Sichuan University, Sichuan, 610041, P.R. China.
Huang
Anliang
A
Yang
Yang
Y
Xie
Gang
G
Chen
Xiang
X
Hu
Jia
J
Chen
Xiancheng
X
Yang
Li
L
Li
Jiong
J
Chen
Lijuan
L
Jiang
Yu
Y
Zhao
Xia
X
Wei
Yuquan
Y
Deng
Hongxin
H
D064127:Aurora Kinases; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000188:drug therapy* / Q000235:genetics*; D002412:Cations; D045744:Cell Line, Tumor; D004247:DNA / Q000378:metabolism*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D005821:Genetic Techniques; D005822:Genetic Vectors; D006801:Humans; D019394:Ki-67 Antigen / Q000096:biosynthesis; D008081:Liposomes / Q000737:chemistry; D009368:Neoplasm Transplantation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002412:Cations; D019394:Ki-67 Antigen; D008081:Liposomes; D004247:DNA; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases