The combination of ANT2 shRNA and hNIS radioiodine gene therapy increases CTL cytotoxic activity through the phenotypic modulation of cancer cells: combination treatment with ANT2 shRNA and I-131

被引:6
作者
Choi, Yun [1 ]
Lee, Ho Won [2 ]
Lee, Jaetae [2 ,3 ]
Jeon, Yong Hyun [2 ,3 ]
机构
[1] Seoul Natl Univ, Coll Med, Dept Pathol, Seoul 151, South Korea
[2] Kyungpook Natl Univ, Dept Nucl Med, Taegu 700721, South Korea
[3] Kyungpook Natl Univ, Leading Edge Res Ctr Drug Discovery & Dev Diabet, Taegu 700721, South Korea
来源
BMC CANCER | 2013年 / 13卷
基金
新加坡国家研究基金会;
关键词
Human sodium iodide symporter (hNIS); Radioiodine gene therapy; Adenine nucleotide translocase-2 (ANT2); Short hairpin RNA (shRNA); Radiation-induced immune response; Cytotoxic T cells (CTLs); SODIUM-IODIDE SYMPORTER; CLASS-I ANTIGENS; TUMOR-CELLS; SODIUM/IODIDE SYMPORTER; T-LYMPHOCYTES; IMMUNOTHERAPY; SIRNA; RADIOTHERAPY; RNA; INTERFERENCE;
D O I
10.1186/1471-2407-13-143
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Background: It is important to simultaneously induce strong cell death and antitumor immunity in cancer patients for successful cancer treatment. Here, we investigated the cytotoxic and phenotypic modulation effects of the combination of ANT2 shRNA and human sodium iodide symporter (hNIS) radioiodine gene therapy in vitro and in vivo and visualized the antitumor effects in an immunocompromised mouse colon cancer model. Methods: A mouse colon cancer cell line co-expressing hNIS and the luciferase gene (CT26/hNIS-Fluc, named CT26/NF) was established. CT26/NF cells and tumor-bearing mice were treated with HBSS, scramble, ANT2 shRNA, I-131, and ANT2 shRNA + I-131. The apoptotic rates (%) and MHC class I and Fas gene expression levels were determined in treated CT26/NF cells using flow cytometry. Concurrently, the level of caspase-3 activation was determined in treated cells in vitro. For in vivo therapy, tumor-bearing mice were treated with scramble, ANT2 shRNA, I-131, and the combination therapy, and the anti-tumor effects were monitored using bioluminescence. The killing activity of cytotoxic T cells (CTLs) was measured with a lactate dehydrogenase (LDH) assay. Results: For the in vitro experiments, the combination of ANT2 shRNA and I-131 resulted in a higher apoptotic cell death rate compared with ANT2 shRNA or I-131 alone, and the levels of MHC class I and Fas-expressing cancer cells were highest in the cells receiving combination treatment, while single treatment modestly increased the level of MHC class I and Fas gene expression. The combination of ANT2 shRNA and I-131 resulted in a higher caspase-3 activation than single treatments. Interestingly, in vivo combination treatment led to increased gene expression of MHC class I and Fas than the respective mono-therapies; furthermore, bioluminescence showed increased antitumor effects after combination treatment than monotherapies. The LDH assay revealed that the CTL killing activity against CT26/NF cells was most effective after combination therapy. Conclusions: Increased cell death and phenotypic modulation of cancer cells in vitro and in vivo were achieved simultaneously after combination therapy with ANT2 shRNA and I-131, and this combination therapy induced remarkable antitumor outcomes through improvements in CTL immunity against CT26/NF. Our results suggest that combination therapy can be used as a new therapeutic strategy for cancer patients who show resistance to single therapy such as radiation or immunotherapy.
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页数:13
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