| 摘要: |
| ［摘要］ 目的 构建耐索拉非尼肝癌细胞株并探讨其耐药机制。方法 应用HepG2、Huh7细胞通过浓度递增法建立耐索拉非尼细胞模型。通过CCK8法检测细胞对不同浓度索拉非尼的敏感性。通过划痕实验检测细胞的迁移能力。通过流式细胞术分析细胞凋亡情况。通过荧光实时定量RT-PCR和Western blot实验检测凋亡相关因子caspase-3和自噬标志因子P62、LC3的表达水平。结果 获得耐索拉非尼肝癌细胞株HepG2-SR、Huh7-SR。随着索拉非尼药物干预浓度的升高，耐药细胞株及野生型细胞株的存活率均逐渐降低。HepG2-SR细胞的半数抑制浓度（IC50）显著高于HepG2细胞［（15.74±0.38）μM vs （7.27±0.44）μM ；t=5.450，P<0.001］；Huh7-SR细胞的IC50显著高于Huh7细胞［（11.73±0.27）μM vs （4.92±0.31）μM；t=4.807，P<0.001］。相较于野生型细胞株，耐药细胞株具有更高的抗凋亡和迁徙能力。耐药细胞株的caspase-3、P62表达水平显著低于野生型细胞株（P<0.05），而LC3表达水平显著高于野生型细胞株（P<0.05）。结论 耐索拉非尼肝癌细胞株HepG2-SR、Huh7-SR具有更强的抗凋亡、抗生殖抑制和迁移能力，且自噬水平更高，为研究肝细胞癌对索拉非尼的耐药机制提供了基础。 |
| 关键词: 索拉非尼 耐药性 肝细胞癌 自噬 |
| DOI：10.3969/j.issn.1674-3806.2023.02.07 |
| 分类号:R 735.7 |
| 基金项目:国家自然科学基金项目（编号：81860654） |
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| Construction of sorafenib-resistant hepatocellular carcinoma cell line and study on its drug-resistant mechanism |
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ZENG Qiong-rong, LIU Ze-feng, WANG Wen-juan, et al.
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Graduate School, Youjiang Medical University for Nationalities, Baise 533000, China
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| Abstract: |
| ［Abstract］ Objective To construct sorafenib-resistant hepatocellular carcinoma cell line and to explore its drug-resistant mechanism. Methods HepG2 and Huh7 cells were used to establish sorafenib-resistant cell models by stepwise increase in drug concentrations method. The sensitivity of cells to sorafenib at different concentrations was detected by CCK8 assay. The migration ability of cells was detected by wound healing assay. The apoptosis of cells was analyzed by flow cytometry. The expression levels of apoptosis-related factor caspase-3 and autophagy marker factors P62 and LC3 were detected by real-time quantitative reverse transcription-polymerase chain reaction(RT-PCR) and Western blot. Results Sorafenib-resistant hepatocellular carcinoma cell lines HepG2-SR and Huh7-SR were obtained.With the increase of sorafenib intervention concentration, the survival rates of drug-resistant cell lines and wild-type cell lines decreased gradually. The half inhibitory concentration(IC50) of HepG2-SR cells was significantly higher than that of HepG2 cells［(15.74±0.38)μM vs (7.27±0.44)μM; t=5.450, P<0.001］. The IC50 of Huh7-SR cells was significantly higher than that of Huh7 cells［(11.73±0.27)μM vs (4.92±0.31)μM; t=4.807, P<0.001］. Compared with the wild-type cell lines, the drug-resistant cell lines showed higher anti-apoptosis and migration ability. The expression levels of caspase-3 and P62 in the drug-resistant cell lines were significantly lower than those in the wild-type cell lines(P<0.05), while the expression level of LC3 in the drug-resistant cell lines was significantly higher than that in the wild-type cell lines(P<0.05). Conclusion Sorafenib-resistant hepatocellular carcinoma cell lines HepG2-SR and Huh7-SR have stronger anti-apoptosis, anti-reproductive inhibition and migration ability, and higher autophagy level, which provides a basis for studying the mechanism of sorafenib resistance in hepatocellular carcinoma. |
| Key words: Sorafenib Drug resistance Hepatocellular carcinoma Autophagy |
