| RGD修饰的肿瘤抑素19肽对SK-Hep-1肝癌细胞增殖及凋亡的影响 |
| 投稿时间：2025-06-06 修订日期：2025-07-23 点此下载全文 |
| 引用本文：王顺,胡月,赵正林,牛淑冬,贾迪,杨超,衣同辉,李淑艳.RGD修饰的肿瘤抑素19肽对SK-Hep-1肝癌细胞增殖及凋亡的影响[J].医学研究杂志,2025,54(12):134-139 |
| DOI：
10.11969/j.issn.1673-548X.2025.12.023 |
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| 基金项目:黑龙江省齐齐哈尔市科学技术局面上项目(LSFGG-2023030) |
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| 中文摘要:目的 研究精氨酸-甘氨酸-天冬氨酸(arginine-glycine-aspartic acid,RGD)修饰的肿瘤抑素19肽(RGD-T-19)对肝癌细胞增殖和凋亡的影响。方法 利用CCK-8实验及克隆形成实验检测T-19和RGD-T-19对SK-Hep-1细胞活力及增殖能力的影响；通过DAPI染色观察细胞凋亡的形态学变化；流式细胞术检测细胞凋亡率；实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)检测天冬氨酸蛋白水解酶caspase-3和caspase-9的mRNA表达水平的变化；Western blot法检测cleaved caspase-3、cleaved caspase-8、cleaved caspase-9和裂解的多腺苷二磷酸核糖聚合酶1(cleaved poly ADP-ribosepolymerase 1, cleaved PARP1)蛋白水平的变化。结果 CCK-8实验结果显示,作用24h和48h后,T-19的半数抑制浓度(half maximal inhibitory concentration, IC50)分别为190.0μg/ml和160.0μg/ml；RGD-T-19的IC50分别为150.0μg/ml和99.0μg/ml。克隆形成实验结果显示,RGD-T-19组的相对细胞克隆数明显少于T-19组(P<0.05)。DAPI染色结果显示,与T-19组比较,RGD-T-19组细胞凋亡形态学变化更加明显；流式细胞术结果显示,RGD-T-19组细胞凋亡率明显大于T-19组(P<0.05)。RT-qPCR和Western blot法检测结果显示,RGD-T-19能显著上调caspase-3和caspase-9的基因表达(P<0.05)和cleaved caspase-3、cleaved caspase-8、cleaved caspase-9和cleaved PARP1的蛋白表达(P<0.05)。结论 RGD-T-19能够明显抑制SK-Hep-1细胞增殖和诱导细胞凋亡。 |
| 中文关键词:RGD 肿瘤抑素 肝癌 增殖 凋亡 |
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| Effect of RGD-T-19 on the Proliferation and Apoptosis of Liver Cancer SK-Hep-1 Cells. |
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| Abstract:Objective To study the effect of arginine-glycine-aspartic acid (RGD)-modified tumstatin 19 peptide (RGD-T-19) on the proliferation and apoptosis of liver cancer cells. Methods CCK-8 assay and clone formation test was used to detect the effects of T-19 and RGD-T-19 on the viability and proliferation of SK-Hep-1 cells. DAPI staining assay was used to observe the morphological changes of apoptosis. Apoptosis rate was detected by flow cytometry. The mRNA expression levels of cysteine aspartate-specific protease caspase-3 and caspase-9 were detected by real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). The protein expression levels of cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, and cleaved poly ADP-ribosepolymerase 1 (cleaved PARP1) were detected by Western blot assay. Results The results of CCK-8 test showed that the half maximal inhibitory concentration (IC50) of T-19 and RGD-T-19 of 24h were 190.0μg/ml and 150.0μg/ml, respectively, the IC50 of T-19 and RGD-T-19 of 48hours were 160.0μg/ml and 99.0μg/ml, respectively. The results of the clone formation test showed that the relative cell clone number of the RGD-T-19 group was significantly less than that of the T-19 group (P<0.05). The results of DAPI staining showed that the morphological changes caused by apoptosis of RGD-T-19 group was more significant than that of T-19 (P<0.05). The results of flow cytometry showed that the apoptosis rate of RGD-T-19 group was higher than that of T-19 group (P<0.05). The results of RT-qPCR and Western blot detection showed that RGD-T-19 can significantly up-regulated the gene expressions of caspase-3 and caspase-9 (P<0.05), and the protein expressions of cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, and cleaved PARP1 (P<0.05). Conclusion The RGD-T-19 can significantly inhibit the proliferation and induce cell apoptosis of SK-Hep-1 cells. |
| keywords:RGD Tumstatin Liver cancer Proliferation Apoptosis |
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