[Effects of heparanase inhibition by RNA interference on proliferation, invasiveness and apoptosis of lung cancer cells].
OBJECTIVE
To investigate the effects of heparanase expression inhibition on the proliferation, invasiveness and apoptosis of human lung adenocarcinoma cell line A549 cells.


METHODS
Recombinant eukaryotic expression plasmid pshRNA-Hpa targeting human heparanase gene was constructed. A549 cells were cultured in DMEM and transfected with pshRNA-Hpa. The expression of heparanase mRNA and protein were examined by RT-PCR and Western blot. The proliferation, invasiveness and apoptotic rates of A549 cells were determined by MTT method, matrigel invasion assays and flow cytometry respectively.


RESULTS
The expression levels of heparanase mRNA and protein were down-regulated in A549 transfected with pshRNA-Hpa. The number of cells penetrating matrigel and the proliferation ability of A549 cells transfected with pshRNA-Hpa were reduced significantly compared to the control cells. The apoptotic rate of A549 cells transfected with pshRNA-Hpa was 12.53% +/- 0.34%, being significantly higher than that of the control cells (both P < 0.01). Western-blot showed that inhibition of heparanase expression led to reduced Akt phosphorylation.


CONCLUSIONS
The recombinant plasmid pshRNA-Hpa effectively inhibited the expression of heparanase, thus suppressing the proliferation and invasion and inducing apoptosis of A549 cells. The effects may be due to the down-regulation of Akt phosphorylation level.
Zhang
Qing-Fu
QF
Department of Pathology, College of Basic Medical Sciences, China Medical University and Department of Pathology, the First Affiliated Hospital of China Medical University, Shenyang 110001, China.
Huang
Qin
Q
Liu
Nan
N
Jiang
Li-Li
LL
Qiu
Xue-Shan
XS
Wang
En-Hua
EH
D000230:Adenocarcinoma / Q000473:pathology; D017209:Apoptosis / Q000187:drug effects*; D002289:Carcinoma, Non-Small-Cell Lung / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D015536:Down-Regulation; D005966:Glucuronidase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000201:enzymology / Q000473:pathology*; D034622:RNA Interference / Q000276:immunology*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology*; D014162:Transfection; D000099175:Heparanase
D005966:Glucuronidase; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D000099175:Heparanase