A novel dithiocarbamate derivative induces cell apoptosis through p53-dependent intrinsic pathway and suppresses the expression of the E6 oncogene of human papillomavirus 18 in HeLa cells
A novel dithiocarbamate derivative induces cell apoptosis through p53-dependent intrinsic pathway and suppresses the expression of the E6 oncogene of human papillomavirus 18 in HeLa cells
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DOI：
10.1007/s10495-015-1114-4
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发表时间：
2015-06-01
期刊：
影响因子：
7.2
通讯作者：
Xiao, Xiangwen
中科院分区：
文献类型：
--
作者：
Li, Yanhong;Qi, Hongxue;Xiao, Xiangwen
二硫代氨基甲酸盐（DTC）具有广谱的抗肿瘤活性，但其抗肿瘤的分子机制尚未阐明。此前，我们合成了一系列新型二硫代氨基甲酸酯衍生物。检查了这些 DTC 对五种人类癌细胞系的细胞毒性活性。本研究选择了一种对HeLa细胞具有较高潜力的二硫代氨基甲酸盐（DTC1）来研究其抗肿瘤活性的分子机制。 DTC1可抑制HeLa细胞增殖，并通过激活caspase-3、-6和-9高度诱导细胞凋亡；此外，Caspase-3、-6和-9的活性被泛Caspase抑制剂Z-VAD-FMK抑制。此外，DTC1以时间依赖性方式降低Bcl-2和Bcl-xL的水平，并增加胞质细胞色素c、Bak、Bax和p53的表达，但对Rb的水平没有影响。野生型 p53 抑制剂 Pifithrin-α 测试显示，DTC1 通过 p53 依赖性途径诱导 HeLa 细胞凋亡。此外，通过实时 PCR 和蛋白质印迹评估了 HPV18 阳性（HeLa 细胞）中 E6 和 E7 的相对表达。结果首先证明DTC1抑制HPV18中E6 mRNA和E6癌蛋白的表达，但对E7 mRNA和蛋白的表达没有影响。我们的结果表明，DTC1 可能作为治疗宫颈癌的新型化疗药物，也是值得进一步研究的潜在抗 HPV 病毒候选药物。
Dithiocarbamates (DTCs) exhibit a broad spectrum of antitumor activities, however, their molecular mechanisms of antitumor have not yet been elucidated. Previously, we have synthesized a series of novel dithiocarbamate derivatives. These DTCs were examined for cytotoxic activities against five human cancer cell lines. In this study, one of dithiocarbamate (DTC1) with higher potential for HeLa cells was chosen to investigate molecular mechanisms for its anti-tumor activities. DTC1 could inhibit proliferation, and highly induce apoptosis in HeLa cells by activating caspase-3, -6 and -9; moreover, activities of caspase-3, -6 and -9 were inhibited by pan-caspase inhibitor, Z-VAD-FMK. Furthermore, DTC1 decreased the levels of Bcl-2 and Bcl-xL, and increased expression of cytosol cytochrome c, Bak, Bax and p53 in a time-dependent manner but had no effect on the level of Rb. It was shown that DTC1 induced HeLa cells apoptosis through a p53-dependent pathway as tested by the wild type p53 inhibitor, pifithrin-alpha. Additionally, the relative expression of E6 and E7 were evaluated in HPV18-positive (HeLa cells) by real-time PCR and western blotting. The results firstly demonstrated that DTC1 suppressed both expression of E6 mRNA and E6 oncoprotein, but had no effect on the expression of E7 mRNA and protein in HPV18. Our results suggested that DTC1 may serve as novel chemotherapeutic agents in the treatment of cervical cancer and potential anti-HPV virus candidates that merit further studies.