Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Senftenberg, Germany.
MVZ Gemeinschaftslabor Cottbus, Cottbus, Germany.
Clin Hemorheol Microcirc. 2024;86(1-2):159-168. doi: 10.3233/CH-238108.
Liver biotransformation is the major route for drug metabolism in humans, often catalysed by cytochrome P450 (CYP) enzymes. This first-pass effect can lead to hepatotoxicity and influences the bioavailability of drugs.
We aimed to establish in vitro culture systems simulating the liver first-pass to study effects of the proteasome inhibitor MG-132 simultaneously on hepatocytes and cancer cells.
The first-pass effect was simulated by conditioned medium transfer (CMT) from pre-treated HepG2 CYP3A4-overexpressing cells to either pancreatic cancer cell line PANC-1 or primary colon cancer cells, and by indirect co-culture (CC) of liver and cancer cells in a shared medium compartment. Experimental proteasome inhibitor MG-132 was used as test substance as it is detoxified by CYP3A4.
Cancer cells showed higher viabilities in the first-pass simulation by CMT and CC formats when compared to monocultures indicating effective detoxification of MG-132 by HepG2 CYP3A4-overexpressing cells. HepG2-CYP3A4 cells showed reduced viabilites after treatment with MG-132.
We successfully established two different culture systems to simulate the liver first-pass effect in vitro. Such systems easily allow to study drug effects simultaneously on liver and on target cancer cells. They are of great value in pre-clinical cancer research, pharmaceutical research and drug development.
肝脏生物转化是人体药物代谢的主要途径，通常由细胞色素 P450（CYP）酶催化。这种首过效应会导致肝毒性，并影响药物的生物利用度。
我们旨在建立模拟肝脏首过效应的体外培养系统，以研究蛋白酶体抑制剂 MG-132 同时对肝细胞和癌细胞的影响。
通过预先用 HepG2 CYP3A4 过表达细胞处理的条件培养基转移（CMT），将首过效应模拟为胰腺癌细胞系 PANC-1 或原代结肠癌细胞，或通过在共享培养基隔室中肝脏和癌细胞的间接共培养（CC）。实验性蛋白酶体抑制剂 MG-132 用作测试物质，因为它被 CYP3A4 解毒。
与单核培养相比，CMT 和 CC 格式的癌细胞在首过模拟中显示出更高的活力，表明 HepG2 CYP3A4 过表达细胞对 MG-132 的有效解毒。用 MG-132 处理后，HepG2-CYP3A4 细胞的活力降低。
我们成功建立了两种不同的体外模拟肝脏首过效应的培养系统。这些系统可轻松用于同时研究药物对肝脏和靶癌细胞的作用。它们在临床前癌症研究、药物研究和药物开发中具有重要价值。
