Le Gac Grégoire, Angenard Gaëlle, Clément Bruno, Laviolle Bruno, Coulouarn Cédric, Beloeil Hélène
From the *INSERM, UMR 991, and Université de Rennes 1, Rennes, France; †CHU Rennes, Pôle Anesthésie et Réanimation, Inserm CIC 1414, Rennes, France; and ‡CHU Rennes, Clinical Pharmacology Department and Inserm CIC 1414, Université de Rennes 1, Rennes, France.
Anesth Analg. 2017 Nov;125(5):1600-1609. doi: 10.1213/ANE.0000000000002429.
Hepatocellular carcinoma (HCC) is an aggressive cancer with limited therapeutic options. Retrospective studies have shown that the administration of local anesthetics (LAs) during cancer surgery could reduce cancer recurrence. Besides, experimental studies reported that LAs could inhibit the growth of cancer cells. Thus, the purpose of this study was to investigate the effects of LAs on human HCC cells.
The effects of 2 LAs (lidocaine and ropivacaine) (10 to 10 M) were studied after an incubation of 48 hours on 2 HCC cell lines, namely HuH7 and HepaRG. Cell viability, cell cycle analysis, and apoptosis and senescence tests were performed together with unsupervised genome-wide expression profiling and quantitative real-time polymerase chain reaction for relevant genes.
We showed that LAs decreased viability and proliferation of HuH7 cells (from 92% [P < .001] at 5 × 10 M to 40% [P = .02] at 10 M with ropivacaine and from 87% [P < .001] to 37% [P = .02] with lidocaine) and HepaRG progenitor cells (from 58% at 5 × 10 M [P < .001] to 29% at 10 M [P = .04] with lidocaine and 59% [P < .001] with ropivacaine 5 × 10 M) in concentration-dependent manner. LAs have no effect on well-differentiated HepaRG. Ropivacaine decreased the mRNA level of key cell cycle regulators, namely cyclin A2, cyclin B1, cyclin B2, and cyclin-dependent kinase 1, and the expression of the nuclear marker of cell proliferation MKI67. Lidocaine had no specific effect on cell cycle but increased by 10× the mRNA level of adenomatous polyposis coli (P < .01), which acts as an antagonist of the Wnt/β-catenin pathway. Both LAs increased apoptosis in Huh7 and HepaRG progenitor cells (P < .01).
The data demonstrate that LAs induced profound modifications in gene expression profiles of tumor cells, including modulations in the expression of cell cycle-related genes that result in a cytostatic effect and induction of apoptosis.
肝细胞癌（HCC）是一种侵袭性癌症，治疗选择有限。回顾性研究表明，癌症手术期间给予局部麻醉剂（LA）可降低癌症复发率。此外，实验研究报告称，LA可抑制癌细胞生长。因此，本研究旨在探讨LA对人肝癌细胞的影响。
在2种肝癌细胞系（即HuH7和HepaRG）上孵育48小时后，研究了2种LA（利多卡因和罗哌卡因）（10至10 M）的作用。进行了细胞活力、细胞周期分析、凋亡和衰老测试，并对相关基因进行了无监督全基因组表达谱分析和定量实时聚合酶链反应。
我们发现，LA以浓度依赖性方式降低了HuH7细胞（罗哌卡因从5×10 M时的92%[P <.001]降至10 M时的40%[P =.02]，利多卡因从87%[P <.001]降至37%[P =.02]）和HepaRG祖细胞（利多卡因从5×10 M时的58%[P <.001]降至10 M时的29%[P =.04]，罗哌卡因5×10 M时为59%[P <.001]）的活力和增殖。LA对分化良好的HepaRG无影响。罗哌卡因降低了关键细胞周期调节因子即细胞周期蛋白A2、细胞周期蛋白B1、细胞周期蛋白B2和细胞周期蛋白依赖性激酶1的mRNA水平，以及细胞增殖核标志物MKI67的表达。利多卡因对细胞周期无特异性影响，但使腺瘤性息肉病大肠杆菌的mRNA水平增加了10倍（P <.01），该基因作为Wnt/β-连环蛋白通路的拮抗剂。两种LA均增加了Huh7和HepaRG祖细胞的凋亡（P <.01）。
数据表明，LA可引起肿瘤细胞基因表达谱的深刻改变，包括细胞周期相关基因表达的调节，从而导致细胞生长抑制作用和凋亡诱导。
