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WANG Yan; DU Liqing; XU Chang; CAO Jia; WANG Qin; CHEN Fenghua; FU Yue; GUO Yanting; LIU Qiang; FAN Feiyue; FAN Saijun

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更新时间：2021-12-01

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- Journal of Radiation Research and Radiation Processing Vol. 31, Issue 6, Pages: 60202-060202(2013)
- 作者机构：
  
  1. （北京协和医学院中国医学科学院放射医学研究所天津市分子核医学重点实验室）,天津,300192
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WANG Yan, DU Liqing, XU Chang, et al. [J]. Journal of Radiation Research and Radiation Processing 31(6):60202-060202(2013)
DOI：

WANG Yan, DU Liqing, XU Chang, et al. [J]. Journal of Radiation Research and Radiation Processing 31(6):60202-060202(2013) DOI：

摘要

探讨Tat-SmacN7诱导食管癌细胞株EC109和非小细胞肺癌细胞株NCL-H460辐射敏感性的机制。采用体外培养EC109细胞和NCL-H460细胞，实验分为对照组、单纯给药组、单纯照射组和给药联合照射组，用RT-PCR检测CAPS8、CASP9和CASP3的转录水平，Fluorogenic caspase assay试剂盒检测Caspase -3、-8、和-9的活性，ELISA方法检测Caspase活性。结果显示，两种肿瘤细胞在使用Tat-SmacN7联合Caspase的抑制剂z-VAD-fmk后，存活曲线较不使用抑制剂组均有明显上移；Caspase -3、-8、和-9的活性在Tat-SmacN7组明显提高，提示Tat-SmacN7可通过细胞凋亡线粒体途径发挥辐射增敏作用，有望成为一种新的辐射增敏剂。

Abstract

To explore the mechanisms of Tat-SmacN7 in radiosensitization by the activation of caspases and induction of apoptosis pathway in EC109 and NCL-H460 cancer cells. The cells were divided into four groups: the control group, the Tat-SmacN7 group, the radiation group, the combined Tat-SmacN7 with radiation group. The transcriptional level of caspase-3, -8 and -9 were fitted by RNA extraction, cDNA synthesis and quantitative real-time PCR. The activity of caspase-3, -8 and-9, was analyzed using a fluorogenic caspase assay with Ac-DEVD-AFC, Ac-IETD-AFC, Ac-LEHD-AFC as substrates, respectively. z-VAD-fmk treatment completely blocked apoptotic combination of Tat-SmacN7 and radiation in both EC109 and H460 cells. The activity and mRNA expression of caspase-3, -8 and -9 were found increased significantly in Tat-SmacN7 group. Tat-SmacN7 increases tumor radiosensitivity through mitochondria pathway of apoptosis, which is expected to be a new radiosensitization agent in the future.Cited

关键词

Tat-SmacN7EC109NCL-H460半胱天冬酶辐射敏感性

Keywords

Tat-SmacN7EC109NCL-H460CaspaseRadiosensitivity

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