| 范冬梅,卢 杨,杨圆圆.可溶性肿瘤坏死因子相关的凋亡诱导配体与5-Fu联用对肝癌细胞的抑制作用[J].中国肿瘤,2017,26(11):904-909. |
| 可溶性肿瘤坏死因子相关的凋亡诱导配体与5-Fu联用对肝癌细胞的抑制作用 |
| Inhibition of Soluble TNF-related Apoptosis-inducing Ligand Combined with 5-Fu on Human Hepatic Carcinoma Cells |
| 投稿时间：2017-03-09 |
| DOI：10.11735/j.issn.1004-0242.2017.11.A012 |
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| 中文关键词: TRAIL 氟尿嘧啶 凋亡相关蛋白 肝癌 |
| 英文关键词:TRAIL fluorouracil apoptosis-related protein hepatocarcinoma |
| 基金项目:国家自然科学基金项目（81400176）；天津市应用基础与前沿技术研究计划（14JCYBJC23700） |
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| 摘要点击次数: 2015 |
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| 中文摘要: |
| 摘 要：［目的］ 探讨肿瘤坏死因子(tumor necrosis factor，TNF)相关的凋亡诱导配体ILZ-sTRAIL与5-氟尿嘧啶（5-fluorouracil，5-Fu）联合应用，对HepG2肝癌细胞生长的抑制作用。［方法］采用聚合酶链反应（PCR）、酶切、连接等方法构建CMV启动子驱动TNF相关凋亡诱导配体（TRAIL）表达的载体pLentiR.ILZ-sTRAIL及其对照载体pLentiR.CopGFP，经测序验证其正确性。将pLentiR.ILZ-sTRAIL和pLentiR.CopGFP分别转染至293T细胞，48h后收集上清并利用ELISA检测目的蛋白含量。利用CCK8细胞增殖抑制实验、Western blot检测ILZ-sTRAIL与5-Fu联用对HepG2肝癌细胞生长的抑制作用及凋亡蛋白的表达变化。［结果］成功构建表达载体pLentiR.ILZ-sTRAIL及pLentiR.CopGFP，测序正确。ILZ-sTRAIL的表达量达（9.475±0.786）ng/ml。体外ILZ-sTRAIL与5-Fu联用，可协同抑制HepG2肝癌细胞的生长。其中凋亡相关蛋白Bcl-2表达降低，Bax表达增加，Caspase-8、9、3及PARP被剪切活化。［结论］ ILZ-sTRAIL蛋白与5-Fu联合应用对肝癌细胞HepG2具有抑制作用，为肝癌的治疗提供了一种新策略。 |
| 英文摘要: |
| Abstract：［Purpose］To investigate the inhibition effects of TNF-related apoptosis-inducing ligand (ILZ-sTRAIL) combined with 5-Fu on human hepatic carcinoma HepG2 cells. ［Methods］ Polymerase chain reaction(PCR)，restriction enzyme cleavage and linkage were used to construct the expression vector pLentiR.ILZ-sTRAIL and its control vector pLentiR.CopGFP. The plasmids were verified by DNA sequencing. pLentiR.ILZ-sTRAIL and pLentiR. CopGFP were transfected to 293T cells，respectively；the supernatants of 293T cells were harvested 48h after transfection and the ILZ-sTRAIL in the supernatants was quantified by ELISA. CCK8 assay was employed to detect the growth inhibition of ILZ-sTRAIL on HepG2 cells when combined with low does of 5-Fu. In addition，the apoptosis-related protein in HepG2 cells were determined by Western blotting. ［Results］ Plasmids pLentiR.ILZ-sTRAIL and pLentiR.CopGFP were successfully constructed and verified by DNA sequencing. The concentration of ILZ-sTRAIL in supernatants reached(9.475±0.786)ng/ml. When treated with ILZ-sTRAIL and 5-Fu in vitro，the growth of HepG2 cells were inhibited significantly. And the expression of Bcl-2 decreased，Bax increased，and caspase-8，9，3 and PARP were cleaved and activated. ［Conclusion］ The combining of ILZ-sTRAIL with 5-Fu can effectively inhibit human hepatic carcinoma HepG2 cells in vitro，which may provide a new approach for treatment of hepatic carcinoma. |
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