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铂纳米胶体联合γ射线辐射对正常人食管上皮细胞的影响

Effects of Platinum Nanocolloid in Combination with Gamma Irradiation on Normal Human Esophageal Epithelial Cells.

作者信息

Li Qiang, Tanaka Yoshiharu, Saitoh Yasukazu, Miwa Nobuhiko

出版信息

J Nanosci Nanotechnol. 2016 May;16(5):5345-52. doi: 10.1166/jnn.2016.12362.

Abstract

Our previous study demonstrated that platinum nanocolloid (Pt-nc), combined with lower-dose gamma irradiation at 3, 5, and 7 Gy significantly decreased proliferation and accelerated apoptosis of the human esophageal squamous cell carcinoma-derived cell line KYSE-70. The aim of the present study was to determine, under the same conditions as our previous study where gamma rays combined with Pt-nc were carcinostatic to KYSE-70 cells, if we could induce a radioprotective or the radiation-sensitizing effect on the human normal esophageal epithelial cells (HEEpiC). HEEpiC were treated with various Pt-nc concentrations and then irradiated with various gamma-ray doses. The proliferative status of HEEpiC was evaluated using trypan blue dye-exclusion and WST-8 assays. The cellular and nucleic morphological features were determined using crystal violet and Hoechst 33342 stainings, respectively. The intracellular level of reactive oxygen species (ROS) in HEEpiC was evaluated with a nitro blue tetrazolium (NBT) assay. The apoptotic status was detected with caspase-3, Bax, and Bcl-2 by Western blotting. Either Pt-nc or gamma irradiation could inhibit the growth of HEEpiC; however, their combined use exerted a significant proliferation-inhibitory effect in a Pt-nc dose-dependent manner than gamma irradiation alone. Pt-nc resulted in radiation sensitization rather than radiation protection on HEEpiC in vitro similar to KYSE-70 cells, when Pt-nc was administrated alone or combined with gamma irradiation. Thus, Pt-nc has an inhibitory effect on cell proliferation, a facilitative effect on apoptosis, and a certain degree of toxicity against HEEpiC.

摘要

我们之前的研究表明,铂纳米胶体(Pt-nc)与3、5和7 Gy的低剂量γ射线联合使用,可显著降低人食管鳞状细胞癌衍生细胞系KYSE-70的增殖并加速其凋亡。本研究的目的是在与我们之前的研究相同的条件下(即γ射线与Pt-nc联合对KYSE-70细胞具有抗癌作用),确定我们是否能对人正常食管上皮细胞(HEEpiC)诱导辐射防护或辐射增敏作用。用不同浓度的Pt-nc处理HEEpiC,然后用不同剂量的γ射线进行照射。使用台盼蓝染料排除法和WST-8检测法评估HEEpiC的增殖状态。分别使用结晶紫染色和Hoechst 33342染色确定细胞和细胞核的形态特征。用硝基蓝四氮唑(NBT)检测法评估HEEpiC中活性氧(ROS)的细胞内水平。通过蛋白质免疫印迹法检测caspase-3、Bax和Bcl-2来检测凋亡状态。单独使用Pt-nc或γ射线均可抑制HEEpiC的生长;然而,与单独使用γ射线相比,它们联合使用时以Pt-nc剂量依赖性方式发挥显著的增殖抑制作用。与KYSE-70细胞类似,当单独使用Pt-nc或与γ射线联合使用时,Pt-nc在体外对HEEpiC产生辐射增敏作用而非辐射防护作用。因此,Pt-nc对细胞增殖具有抑制作用,对凋亡具有促进作用,并且对HEEpiC具有一定程度的毒性。

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