[Effect of RNA interference mediated gene silencing of DJ-1 on proliferation of laryngeal carcinoma cell line Hep-2].
OBJECTIVE
To assess the effect of small interfering RNA (siRNA)-mediated gene silencing of DJ-1 on the proliferation of human laryngeal carcinoma cell line Hep-2.


METHODS
Three siRNA sequences specific to DJ-1 gene were synthesized according to GenBank. Human laryngeal carcinoma cell line Hep-2 was cultured and divided into 4 groups: non-specific group (siRNA control) and 3 RNAi groups, transfected with specific DJ-1 siRNA (siRNA1, siRNA2, siRNA3). The mRNA and protein levels of DJ-1 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot respectively. Cell apoptosis were analyzed by flow cytometry. The proliferation of Hep-2 cells was assessed by MTT assay.


RESULTS
DJ-1 siRNA down-regulated the mRNA and protein levels of DJ-1 in Hep-2 cells. After transfection, the expression of DJ-1 mRNA and protein levels in Hep-2 cells of the DJ-1 siRNA1 group were significantly lower than those of non-specific siRNA control group. MTT assay showed that DJ-1 siRNA1 group inhibited proliferation of Hep-2 cells. Flow cytometry showed that apoptosis rate of the DJ-1 siRNA1 group (15.7%) was significantly higher than that of non-specific siRNA control group (4.5%) or untransfected group (3.5%), t = 4.736, P < 0.01.


CONCLUSIONS
Specific siRNA targeting DJ-1 can effectively inhibit DJ-1 expression, resulting in the reduced proliferation and the enhanced apoptosis in Hep-2 cells.
Zhu
Xiao-Lin
XL
Otorhinolaryngology Hospital, First Affiliated Hospital, Otorhinolaryngology Institute of Sun Yat-sen University, Guangzhou 510080, China.
Wen
Wei-Ping
WP
Jiang
Ai-Yun
AY
Lei
Wen-Bin
WB
Su
Zhen-Zhong
ZZ
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics*; D007822:Laryngeal Neoplasms / Q000235:genetics* / Q000473:pathology*; D015513:Oncogene Proteins / Q000235:genetics*; D000071617:Protein Deglycase DJ-1; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics
D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1