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Published on: October 16, 2017
Effect of small interference RNA targeting HIF-1alpha mediated by rAAV combined L: -ascorbate on pancreatic tumors in
Chuangui Chen1, Jinjin Sun, Geng Liu
1Department of Surgery, The Second Hospital of TianJin Medical University, Tianjin, 300211, China.
Abstract:
To study the effect of recombinant adeno-associated virus (rAAV) vector bearing small inference RNA (siRNA) targeting hypoxia inducible factor 1alpha (HIF-1alpha) combined L: -ascorbate on pancreatic tumors in athymic mice primarily. A cassette encoding siRNA targeting HIF-1alpha mediated by rAAV was constructed, giving rAAV-siHIF. In vitro, rAAV-hrGFP, rAAV-siHIF and L: -ascorbate which were used alone or in combination were delivered to exponentially growing MiaPaCa2 cells. Then, we examined the expression of HIF-1alpha mRNA and protein, the secretion of VEGF in MiaPaCa2 cells under hypoxic condition with Real-time PCR, Western Blot, ELISA, respectively. In vivo, MiaPaCa2 cells were inoculated subcutaneously on the back of nude mice. Nude mice with xenograft tumor were randomly divided into equal groups and were injected with rAAV-hrGFP or rAAV-siHIF or were fed with L: -ascorbate. Then, we measured the size of tumor every 3 days and drew a tumor growth curve. After 30 days, all mice were sacrificed and the tumors were dissected. At last, we examined the expression of HIF-1alpha, VEGF and CD34 by immunohistochemistry and counted micro-vessel density (MVD). In vitro, we found that rAAV-siHIF could inhibit the expression of HIF-1alpha mRNA and protein in MiaPaCa2 human pancreatic cancer cells but L: -ascorbate could only restrain the expression of HIF-1alpha protein. Moreover, rAAV-siHIF and L: -ascorbate could all inhibit the secretion of vascular VEGF. In vivo, we found that rAAV-siHIF could inhibit the growth of nude mice xenograft tumor and the expression of HIF-1alpha and VEGF and MVD while L: -ascorbate can only inhibit the growth of xenograft tumor in the early and middle stage. These results suggest that rAAV-siHIF and L: -ascorbate can inhibit the growth of nude mice xenograft tumor and HIF-1alpha could be a target of pancreatic cancer genetic and pharmacological therapy.
Insights
Recombinant adeno-associated virus (rAAV) carrying siRNA targeting hypoxia-inducible factor 1-alpha (HIF-1alpha) and L-ascorbate effectively inhibit pancreatic tumor growth in mice. HIF-1alpha is a promising target for pancreatic cancer therapy.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Pancreatic cancer is a leading cause of cancer-related deaths.
- Hypoxia-inducible factor 1-alpha (HIF-1alpha) is a key regulator of tumor growth and angiogenesis.
- Targeting HIF-1alpha presents a potential therapeutic strategy for pancreatic cancer.
Purpose of the Study:
- To investigate the efficacy of a recombinant adeno-associated virus (rAAV) vector encoding siRNA against HIF-1alpha (rAAV-siHIF), combined with L-ascorbate, in inhibiting pancreatic tumor growth.
- To evaluate the molecular mechanisms underlying the anti-tumor effects of rAAV-siHIF and L-ascorbate in vitro and in vivo.
Main Methods:
- Construction of rAAV vector expressing siRNA targeting HIF-1alpha (rAAV-siHIF).
- In vitro studies using MiaPaCa2 cells treated with rAAV-siHIF and/or L-ascorbate to assess HIF-1alpha expression and VEGF secretion.
- In vivo studies using nude mice xenograft models to evaluate tumor growth, HIF-1alpha, VEGF, and micro-vessel density (MVD) after treatment.
Main Results:
- rAAV-siHIF significantly inhibited HIF-1alpha mRNA and protein expression in MiaPaCa2 cells.
- Both rAAV-siHIF and L-ascorbate reduced VEGF secretion in vitro.
- In vivo, rAAV-siHIF suppressed tumor growth, HIF-1alpha expression, and MVD, while L-ascorbate showed efficacy in early to middle stages of tumor growth.
Conclusions:
- rAAV-siHIF demonstrates potent anti-tumor activity against pancreatic cancer by targeting HIF-1alpha.
- Combination therapy with rAAV-siHIF and L-ascorbate shows promise for pancreatic cancer treatment.
- HIF-1alpha is a viable therapeutic target for genetic and pharmacological interventions in pancreatic cancer.
