JS-K对人胃癌细胞的抑瘤效应及其相关机制的研究
发布时间：2018-07-13 17:33
【摘要】：背景:胃癌是我国的一种常见的消化道恶性肿瘤,由于我国人口众多且经济社会发展的不均衡,我国早期胃癌的就诊比例比较低,而进展期胃癌的比例很高,这严重降低了患者的5年生存率。目前针对进展期胃癌的治疗模式主要是D2根治术联合术后化疗。由于临床常用的化疗药物副作用较大且作用不确定,临床急需新的治疗药物。JS-K是一种新和成的一氧化氮(NO)前体药,研究显示JS-K对多种肿瘤都具有良好的抗肿瘤效应,但JS-K在胃癌中的研究尚未见诸报道。方法:在本研究中,我们通过一系列的体外试验和体内试验,对JS-K在胃癌中的抗肿瘤效应进行评估,并对其具体的作用机制进行探讨。我们通过MTT试验、流式细胞仪检测技术、线粒体分离技术、目的蛋白的质粒过表达、siRNA、Western blot以及胃癌的皮下移植瘤模型等一系列的生化检测手段对上述问题进行研究。结果:JS-K可以通过诱导细胞凋亡对胃癌细胞进行杀伤,且杀伤效果呈时间和剂量依赖性。JS-K诱导胃癌细胞发生的凋亡是caspase依赖性的,泛caspase抑制剂Z-VAD-FMK可以完全阻止JS-K上述的杀伤效果,而caspase3的抑制剂Z-DEVD-FMK和caspase 9抑制剂Z-LEHD-FMK只能部分地阻止JS-K的杀伤效果。JS-K通过抑制呼吸链复合体1 (ComplexⅠ)和复合体Ⅳ (Complex Ⅳ)使胃癌细胞活性氧(ROS)的产生增多,同时又降低SOD1和Catalase这两种活性氧(ROS)清除蛋白的含量,使活性氧(ROS)在胃癌细胞内累积。细胞内累积的活性氧(ROS)损伤线粒体的功能,使线粒体膜电位下降(△Ψm),细胞色素C由线粒体进入细胞质,激活caspase 9,而活化的caspase 9又激活caspase 3和剪切PARP从而导致凋亡的发生。JS-K通过活性氧(ROS)使胃癌细胞发生凋亡,而活性氧清除剂N-乙酰-L-半胱氨酸(NAC)可以逆转JS-K的上述作用。JS-K可以降低胃癌细胞内的抗凋亡蛋白Bcl-2和Bcl-xL的含量,而通过质粒过表达Bcl-2和Bcl-xL这两种蛋白可以保护胃癌细胞免受JS-K的杀伤。通过小干扰RNA (siRNA)技术,我们发现凋亡诱导因子(AIF)和核算内切酶G (Endo G)的核转位并不参与JS-K诱导的胃癌细胞发生的凋亡过程。在体内试验中,JS-K可以很好抑制胃癌皮下瘤体积和质量的增长,但对小鼠的体重并不产生明显的影响,说明其生物耐受性良好。结论:JS-K可以通过使胃癌细胞累积活性氧(ROS)而损伤线粒体,通过线粒体途径使胃癌细胞发生caspase依赖性的凋亡,从而发挥抗肿瘤效应,且其生物耐受性良好,是一种良好的潜在的可以用于治疗胃癌的药物,值得在今后的临床应用和试验中进一步的研究。
[Abstract]:Background: gastric cancer is a common malignant tumor of digestive tract in China. Due to the large population and unbalanced economic and social development, the proportion of patients with early gastric cancer in China is low, but the proportion of advanced gastric cancer is very high. This severely reduces the 5-year survival rate. At present, the main treatment mode for advanced gastric cancer is D 2 radical resection combined with postoperative chemotherapy. Because the side effects of the commonly used chemotherapeutic drugs are large and uncertain, JS-K is a new precursor of nitric oxide (no), which is in urgent need of clinical treatment. Studies show that JS-K has a good anti-tumor effect on many kinds of tumors. However, the study of JS-K in gastric cancer has not been reported. Methods: in this study, we evaluated the antitumor effect of JS-K in gastric cancer by a series of experiments in vitro and in vivo, and discussed its mechanism. We studied the above problems by a series of biochemical methods, such as MTT test, flow cytometry, mitochondrial isolation, plasmid overexpression siRNA blot and subcutaneous transplanted tumor model of gastric cancer. Results the cell apoptosis of gastric cancer cells was induced by 1: JS-K in a time-and dose-dependent manner. The apoptosis of gastric cancer cells induced by JS-K was caspase dependent. Z-VAD-FMK, a pan-caspase inhibitor, could completely prevent the killing effect of JS-K. Z-DEVD-FMK, an inhibitor of caspase3, and Z-LEHD-FMK, an inhibitor of caspase 9, could partially prevent the cytotoxicity of JS-K. JS-K inhibited the production of reactive oxygen species (Ros) in gastric cancer cells by inhibiting the production of complex I and complex 鈪，
本文编号：2120189
[Abstract]:Background: gastric cancer is a common malignant tumor of digestive tract in China. Due to the large population and unbalanced economic and social development, the proportion of patients with early gastric cancer in China is low, but the proportion of advanced gastric cancer is very high. This severely reduces the 5-year survival rate. At present, the main treatment mode for advanced gastric cancer is D 2 radical resection combined with postoperative chemotherapy. Because the side effects of the commonly used chemotherapeutic drugs are large and uncertain, JS-K is a new precursor of nitric oxide (no), which is in urgent need of clinical treatment. Studies show that JS-K has a good anti-tumor effect on many kinds of tumors. However, the study of JS-K in gastric cancer has not been reported. Methods: in this study, we evaluated the antitumor effect of JS-K in gastric cancer by a series of experiments in vitro and in vivo, and discussed its mechanism. We studied the above problems by a series of biochemical methods, such as MTT test, flow cytometry, mitochondrial isolation, plasmid overexpression siRNA blot and subcutaneous transplanted tumor model of gastric cancer. Results the cell apoptosis of gastric cancer cells was induced by 1: JS-K in a time-and dose-dependent manner. The apoptosis of gastric cancer cells induced by JS-K was caspase dependent. Z-VAD-FMK, a pan-caspase inhibitor, could completely prevent the killing effect of JS-K. Z-DEVD-FMK, an inhibitor of caspase3, and Z-LEHD-FMK, an inhibitor of caspase 9, could partially prevent the cytotoxicity of JS-K. JS-K inhibited the production of reactive oxygen species (Ros) in gastric cancer cells by inhibiting the production of complex I and complex 鈪，
本文编号：2120189
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