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Published on: December 15, 2010
Induced apoptosis with ultrasound-mediated microbubble destruction and shRNA targeting survivin in transplanted
Zhi-Yi Chen1, Kun Liang, Ming-Xing Xie
1Department of Medical Ultrasound, The Third Affiliated Hospital of Guangzhou Medical College, Guangdong, 510150, China. winchen1408@126.com
Introduction:
This study was designed to evaluate the consequences of survivin down-modulation on tumor growth in a nude mice model combined with short hairpin RNA recombinant vector (shRNA) and ultrasound-mediated microbubble destruction (UMMD).
Methods:
BALB/c nude mice were inoculated subcutaneously with cervical cancer cells (HeLa) and tumors (5-10 mm) developed. A shRNA recombinant vector that targeted the survivin gene (survivin-shRNA) was constructed. The mice were divided into three groups (n=6 in each group) and injected with survivin-shRNA: plasmid group (P), plasmid+ultrasound exposure group (P+US), and plasmid+microbubble (SonoVue(R))+ultrasound group (P+UMMD). Protein expression of survivin, proliferating cell nuclear antigen (PCNA), and caspase-3 were investigated by immunohistochemistry, and proliferation index (PI) and apoptotic index (AI) were measured.
Results:
The protein expression of survivin and PCNA was markedly downregulated, while caspase-3 was markedly upregulated in the P+UMMD group as compared with that of the P group and P+US group. PI decreased significantly (P<0.05), whereas AI increased remarkably (P<0.01) in the P+UMMD group as compared with that of the P group and P+US group. These data indicate that the combined strategy of UMMD and survivin-shRNA effectively induces silencing of the survivin gene, resulting in inhibition of proliferation and induction of apoptosis in nude mice.
Conclusions:
Survivin could be regarded as an ideal target for anticancer intervention of cervical cancer. The combination of shRNA and UMMD could enhance antitumor efficacy as a result of synergism. This may be a powerful, promising non-viral technology that could be used in tumor gene therapy.
Insights
Combining ultrasound-mediated microbubble destruction (UMMD) with survivin short hairpin RNA (shRNA) effectively suppresses cervical cancer growth in mice. This synergistic approach inhibits tumor proliferation and promotes apoptosis, offering a promising non-viral gene therapy strategy.
Area of Science:
- Oncology
- Gene Therapy
- Biotechnology
Background:
- Cervical cancer remains a significant health concern, necessitating novel therapeutic strategies.
- Survivin is a key protein implicated in tumor growth and resistance to apoptosis.
- Targeting survivin offers a potential avenue for effective cancer intervention.
Purpose of the Study:
- To evaluate the efficacy of survivin down-modulation using short hairpin RNA (shRNA) combined with ultrasound-mediated microbubble destruction (UMMD).
- To assess the impact of this combined therapy on tumor growth, proliferation, and apoptosis in a nude mouse model of cervical cancer.
Main Methods:
- Cervical cancer cells (HeLa) were implanted in nude mice to establish tumors.
- A survivin-specific shRNA recombinant vector was constructed and administered.
- Mice were divided into groups receiving plasmid, plasmid plus ultrasound, or plasmid plus UMMD.
- Survivin, PCNA, and caspase-3 protein expression, along with proliferation and apoptotic indices, were analyzed.
Main Results:
- The combination of UMMD and survivin-shRNA significantly downregulated survivin and PCNA expression.
- Caspase-3 expression was markedly upregulated in the UMMD group.
- Tumor proliferation index decreased, while the apoptotic index increased significantly in the UMMD group.
Conclusions:
- Survivin is a viable molecular target for cervical cancer therapy.
- The synergistic combination of shRNA and UMMD enhances antitumor efficacy.
- This non-viral technology presents a promising approach for tumor gene therapy.
