Silencing Id-1 with RNA interference inhibits adenoid cystic carcinoma in mice
Zhenggang Chen1, Shaohua Liu, Tomoki Sumida
1School of Stomatology, Shandong University, Jinan, People's Republic of China.
Background:
The helix-loop-helix (HLH) protein Id-1 (inhibitor of DNA binding/differentiation) has been demonstrated to play an important role in tumor development. Our previous in vitro research has shown that Id-1 is a potential target in the treatment of human adenoid cystic carcinoma (ACCM). The purpose of this study was to analyze the influence of Id inhibition on ACCM in mice.
Materials And Methods:
To suppress the expression of Id-1 gene, we used lentivirus-mediated RNA interference to silence the Id-1 gene post-transcriptionally in ACCM models that stably express GFP in mice. Tumor development was evaluated by size measurement. Effects of Id-1 siRNA on mRNA and protein expression of Id-1 were analyzed using quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting respectively. Ki-67 expression was measured by immunohistochemistry. In vitro studies of Hoechst staining for cell apoptosis, Boyden-chamber assay for cell invasion, and MTT-tests for cell growth were performed as well.
Results:
Id-1 knockdown resulted in inhibition of tumor growth in mice. Id-1 siRNA significantly decreased not only Id-1 in mRNA and protein level, but also Ki-67 expression. In addition, apoptosis was induced and cell proliferation activity and invasion were significantly reduced.
Conclusions:
Lentivirus-mediated gene knockdown by silencing Id-1 constitute a valid methodological approach, which may represent an attractive, potent and specific therapeutic tool for the treatment of ACCM.
Insights
Silencing the Id-1 gene in mice with adenoid cystic carcinoma (ACCM) inhibited tumor growth. This Id-1 gene knockdown reduced proliferation and invasion, showing potential for ACCM treatment.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- The helix-loop-helix (HLH) protein Id-1 is implicated in tumor development.
- Previous in vitro studies identified Id-1 as a potential therapeutic target for human adenoid cystic carcinoma (ACCM).
Purpose of the Study:
- To investigate the therapeutic potential of Id-1 inhibition in a mouse model of ACCM.
- To evaluate the effects of suppressing Id-1 expression on ACCM progression in vivo.
Main Methods:
- Lentivirus-mediated RNA interference was employed to silence the Id-1 gene in ACCM mouse models.
- Tumor growth was monitored, and Id-1, Ki-67, mRNA, and protein expression levels were analyzed.
- In vitro assays assessed cell apoptosis, invasion, and proliferation.
Main Results:
- Id-1 gene knockdown significantly inhibited tumor growth in mice.
- Reduced Id-1 and Ki-67 expression levels were observed post-silencing.
- Apoptosis was induced, while cell proliferation and invasion were markedly decreased.
Conclusions:
- Lentivirus-mediated Id-1 gene silencing is a viable strategy for treating ACCM.
- This approach demonstrates potential as a potent and specific therapeutic tool for ACCM.
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