siRNA Targeting HIF-1 alpha Induces Apoptosis of Pancreatic Cancer Cells through NF-kappa B-independent and -dependent Pathways under Hypoxic Conditions

作者：Chen Chuangui; Yu Zhentao^*

来源：Anticancer Research, 2009, 29(4): 1367-1372.

• Recombinant adeno-associated virus (rAAV)
• hypoxia inducible factor (HIF)
• small interference RNA (siRNA)
• nuclear factor-kappaB (NF-kappa B)
• proliferation
• apoptosis

Objective: The primary objective was to explore the molecular mechanism of small interference RNA (siRNA) targeting hypoxic inducible factor-1 alpha (HIF-1 alpha) for inducing apoptosis of pancreatic cancer cells through a nuclear factor-kappaB (NF-kappa B)-independent or -dependent pathway tinder hypoxic conditions. Materials and Methods: A cassette encoding siRNA targeting HIF-1 alpha mediated by recombinant adeno-associated virus (rAAV) was constructed, giving rAAV-siHIF. rAAV-siHIF or rAAV-hrGFP were transfected into exponentially growing MiaPaCa2 cells under hypoxic conditions. The expression of HIF-1 alpha mRNA and protein and the activity of NF-kappa B were observed by, real-time PCR, Western blot and electromobility shift assay (EMSA). The proliferation and apoptosis of MiaPaCa2 cells with or without PDTC, as an inhibitor of NF-kappa B, were investigated by MTT and TUNEL. Results: rAAV-siHIF inhibited the expression of HIF-1 alpha mRNA and protein and the activity of NF-kappa B in MiaPaCa2 cells under hypoxic conditions. At the same time, rAAV-siHIF decreased MiaPaCa2 cell proliferation and induced apoptosis, but these effects were not abrogated by PDTC. Moreover, PDTC also inhibited MiaPaCa2 cell proliferation and induced apoptosis while rAAV-hrGFP did not have these effects. Conclusion: Under hypoxic conditions, HIF-1 alpha plays a key role in the Proliferation of MiaPaCa2 cells and inhibition of HIF-1 alpha expression may lead to MiaPaCa2 cell apoptosis through NF-kappa B-independent and -dependent pathways.

• 出版日期2009-4
• 单位天津医科大学