Irie Nanami, Warita Katsuhiko, Tashiro Jiro, Zhou Yaxuan, Ishikawa Takuro, Oltvai Zoltán N, Warita Tomoko
Graduate School of Science and Technology, Kwansei Gakuin University, 1 Gakuen Uegahara, Sanda, Hyogo 669-1330, Japan.
Department of Veterinary Anatomy, School of Veterinary Medicine, Tottori University, 4-101 Koyama Minami, Tottori, Tottori 680-8553, Japan.
Heliyon. 2023 Jul 8;9(7):e18017. doi: 10.1016/j.heliyon.2023.e18017. eCollection 2023 Jul.
Statins have anticancer effects and may be used as anticancer agents via drug repositioning. In reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assays, the internal reference gene must not be affected by any experimental conditions. As statins exert a wide range of effects on cells by inhibiting the mevalonate pathway, it is possible that statin treatment might alter the expression of housekeeping genes used as internal reference genes, thereby misleading the assessment of obtained gene expression data. Here, we evaluated the expression stability of internal reference genes in atorvastatin-treated cancer cell lines. We treated both statin-sensitive and statin-resistant cancer cell lines with atorvastatin at seven different concentrations and performed RT-qPCR on 15 housekeeping genes whose expression stability was then assessed using five different algorithms. In both statin-sensitive and statin-resistant cancer cell lines, atorvastatin affected the expression of certain internal reference genes in a dose-dependent and cancer cell line-dependent manner; therefore, caution should be exercised when comparing target gene expression between cells. Our findings emphasize the importance of the validation of internal reference genes in gene expression analyses in drug treatment-based cancer research.
他汀类药物具有抗癌作用，可通过药物重新定位用作抗癌剂。在逆转录定量聚合酶链反应（RT-qPCR）分析中，内参基因不得受任何实验条件的影响。由于他汀类药物通过抑制甲羟戊酸途径对细胞产生广泛影响，他汀类药物治疗可能会改变用作内参基因的管家基因的表达，从而误导对所获得基因表达数据的评估。在此，我们评估了阿托伐他汀处理的癌细胞系中内参基因的表达稳定性。我们用七种不同浓度的阿托伐他汀处理他汀类药物敏感和耐药的癌细胞系，并对15个管家基因进行RT-qPCR，然后使用五种不同算法评估其表达稳定性。在他汀类药物敏感和耐药的癌细胞系中，阿托伐他汀均以剂量依赖性和癌细胞系依赖性方式影响某些内参基因的表达；因此，在比较细胞间靶基因表达时应谨慎。我们的研究结果强调了在基于药物治疗的癌症研究中基因表达分析中验证内参基因的重要性。
