丙泊酚通过诱导自噬激活AMPK，从而抑制HepG2细胞的生长以及异种移植小鼠肿瘤模型中的肝癌发生。
Propofol activates AMPK to inhibit the growth of HepG2 cells and hepatocarcinogenesis in xenograft mouse tumor models by inducing autophagy.
作者信息
Wang Yixiong, Xu Baozhu, Zhou Jianying, Wu Xianyan
机构信息
Department of Anesthesiology, The Quanzhou First Affiliated Hospital of Fujian Medical University, Quanzhou, China.
Department of Radiology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, China.
出版信息
J Gastrointest Oncol. 2020 Dec;11(6):1322-1332. doi: 10.21037/jgo-20-472.
BACKGROUND
Hepatocellular carcinoma (HCC) is a fatal malignant tumor with a poor prognosis, and is the third leading cause of cancer-related deaths worldwide. This study aimed to investigate the anti-tumor effect of propofol on the proliferation, apoptosis, and cell cycle of HCC by regulating adenosine monophosphate-activated protein kinase (AMPK) and .
METHODS
The cell counting Kit-8 (CCK-8) assay was employed to screen the effect of propofol on HepG2 cell viability at various concentrations (0.3, 0.6, 1.2, 2.5, 5, 10, 20, 40, 80 and 160 µM). We selected propofol at concentrations of 5, 10 and 20 µM for subsequent experiments. Flow cytometry was used to examine the apoptosis and cell cycle of HCC. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was applied to measure the messenger ribonucleic acid (mRNA) expression levels of proliferating cell nuclear antigen (PCNA) and survivin. Western blotting was applied to measure the protein expression levels of PCNA, survivin, cleaved caspase-3, cleaved caspase-9, p27 (Kip1), and cyclin A. The effects of propofol were evaluated by establishing a xenograft tumor model.
RESULTS
After treatment with propofol, the mRNA expression levels of PCNA and survivin were decreased compared with the 0 µM propofol (control) group. The colony formation assay showed that the colony formation rate was obviously down-regulated. Flow cytometry demonstrated that HepG2 cell apoptosis was increased. G0/G1 was enhanced compared with the control group, while G2/M was restrained. The levels of cleaved caspase-3, cleaved caspase-9, p27, phospho-AMP-activated protein kinase α1 (p-AMPKα1), phospho-mammalian target of rapamycin (p-mTOR), and phospho-Unc-51 like autophagy activating kinase 1 (p-ULK1) were notably elevated, while the levels of cyclin A were suppressed. The xenograft tumor volume declined compared with the HepG2 xenograft group. The expression levels of cell proliferation markers (PCNA) were significantly down-regulated markedly, while the expression levels of cell cycle markers (p27) were notablyup-regulated. Terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining showed that cell apoptosis was increased. The levels of p-AMPKα1 were also up-regulated.
CONCLUSIONS
Propofol inhibits the proliferation, apoptosis, and cell cycle of HCC by regulating AMPK and .
背景
肝细胞癌（HCC）是一种预后较差的致命性恶性肿瘤，是全球癌症相关死亡的第三大主要原因。本研究旨在通过调节腺苷单磷酸激活蛋白激酶（AMPK）等来探讨丙泊酚对肝癌细胞增殖、凋亡和细胞周期的抗肿瘤作用。
方法
采用细胞计数试剂盒-8（CCK-8）法检测不同浓度（0.3、0.6、1.2、2.5、5、10、20、40、80和160μM）丙泊酚对HepG2细胞活力的影响。我们选择5、10和20μM浓度的丙泊酚用于后续实验。采用流式细胞术检测肝癌细胞的凋亡和细胞周期。应用定量实时逆转录-聚合酶链反应（qRT-PCR）检测增殖细胞核抗原（PCNA）和生存素的信使核糖核酸（mRNA）表达水平。采用蛋白质印迹法检测PCNA、生存素、裂解的半胱天冬酶-3、裂解的半胱天冬酶-9、p27（Kip1）和细胞周期蛋白A的蛋白表达水平。通过建立异种移植肿瘤模型评估丙泊酚的作用。
结果
与0μM丙泊酚（对照组）相比，丙泊酚处理后PCNA和生存素的mRNA表达水平降低。集落形成试验表明集落形成率明显下调。流式细胞术显示HepG2细胞凋亡增加。与对照组相比，G0/G1期增强，而G2/M期受到抑制。裂解的半胱天冬酶-3、裂解的半胱天冬酶-9、p27、磷酸化腺苷单磷酸激活蛋白激酶α1（p-AMPKα1）、磷酸化雷帕霉素哺乳动物靶蛋白（p-mTOR）和磷酸化Unc-51样自噬激活激酶1（p-ULK1）水平显著升高，而细胞周期蛋白A水平受到抑制。与HepG2异种移植组相比，异种移植肿瘤体积减小。细胞增殖标志物（PCNA）的表达水平显著下调，而细胞周期标志物（p27）的表达水平显著上调。末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）染色显示细胞凋亡增加。p-AMPKα1水平也上调。
结论
丙泊酚通过调节AMPK等来抑制肝癌细胞的增殖、凋亡和细胞周期。
Zotero 插件