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.
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.
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DOI：
10.1186/1471-2407-13-143
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发表时间：
2013-03-22
期刊：
影响因子：
3.8
通讯作者：
Jeon YH
中科院分区：
文献类型：
--
作者：
Choi Y;Lee HW;Lee J;Jeon YH
同时诱导癌症患者强烈的细胞死亡和抗肿瘤免疫对于成功的癌症治疗非常重要。在这里，我们研究了细胞毒性和表型调制作用的组合ANT 2 shRNA和人钠碘同向转运体（hNIS）放射性碘基因治疗在体外和体内，并可视化的抗肿瘤作用在免疫功能低下的小鼠结肠癌模型。建立了共表达hNIS和荧光素酶基因的小鼠结肠癌细胞系（CT 26/hNIS-Fluc，简称CT 26/NF）。用HBSS、scramble、ANT 2 shRNA、I-131和ANT 2 shRNA + I-131处理CT 26/NF细胞和荷瘤小鼠。用流式细胞仪检测CT 26/NF细胞凋亡率（%）、MHC I类分子和Fas基因表达水平。同时，在体外测定经处理的细胞中半胱天冬酶-3活化的水平。对于体内治疗，荷瘤小鼠接受scramble、ANT 2 shRNA、I-131和联合治疗，并使用生物发光监测抗肿瘤效果。用乳酸脱氢酶（LDH）测定法测定细胞毒性T细胞（CTL）的杀伤活性。对于体外实验，与单独的ANT 2 shRNA或I-131相比，ANT 2 shRNA和I-131的组合导致更高的凋亡细胞死亡率，并且在接受组合处理的细胞中表达MHC I类和Fas的癌细胞的水平最高，而单一处理适度地增加了MHC I类和Fas基因表达的水平。ANT 2 shRNA和I-131的组合导致比单一处理更高的caspase-3活化。有趣的是，体内联合治疗导致MHC I类和Fas的基因表达比各自的单一疗法增加;此外，生物发光显示联合治疗后的抗肿瘤作用比单一疗法增加。LDH活性测定结果显示，联合治疗后，CTL对CT 26/NF细胞的杀伤活性最强。在用ANT 2 shRNA和I-131联合治疗后，在体外和体内同时实现了增加的细胞死亡和癌细胞的表型调节，并且这种联合治疗通过改善针对CT 26/NF的CTL免疫而诱导显著的抗肿瘤结果。我们的研究结果表明，联合治疗可以作为一种新的治疗策略，用于对单一治疗（如放射或免疫治疗）表现出耐药性的癌症患者。
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. 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. 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. 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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影响因子：
5.7
作者：
Jeon, Yong Hyun;Choi, Yun;Chung, June-Key
通讯作者：
Chung, June-Key
影响因子：
4.4
作者：
Chakraborty, M;Abrams, SI;Hodge, JW
通讯作者：
Hodge, JW
影响因子：
4.2
作者：
Rosenberg, SA;Yang, JC;White, DE
通讯作者：
White, DE
DOI：
10.1186/bcr1857
发表时间：
2008
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Jang JY;Choi Y;Jeon YK;Kim CW
通讯作者：
Kim CW
影响因子：
11.5
作者：
Gulley, JL;Arlen, PM;Dahut, W
通讯作者：
Dahut, W