Kasi Reena A P, Moi Chye Soi, Kien Yip Wai, Yian Koh Rhun, Chin Ng Wei, Yen Ng Khuen, Ponnudurai Gnanajothy, Fong Seow Heng
Department of Human Biology, Cells and Molecules, International Medical University, Kuala Lumpur 57000, Malaysia.
Department of Pathology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor 43400, Malaysia.
Mol Med Rep. 2015 Mar;11(3):2262-8. doi: 10.3892/mmr.2014.2979. Epub 2014 Nov 19.
para‑Phenylenediamine (p‑PD) is a potential carcinogen, and widely used in marketed hair dye formulations. In the present study, the role of the protein tyrosine kinase (PTK)/Ras/Raf/c‑Jun N‑terminal kinase (JNK) and phosphoinositide 3‑kinase (PI3k)/protein kinase B (Akt) pathways on the growth of NRK‑52E cells was investigated. The results demonstrated that p‑PD reduced cell viability in a dose‑dependent manner. The cell death due to apoptosis was confirmed by cell cycle analysis and an Annexin‑V‑fluorescein isothiocyanate binding assay. Subsequent to staining with 2',7'‑dichlorofluorescin diacetate, the treated cells demonstrated a significant increase in reactive oxygen species (ROS) generation compared with the controls. The effects of p‑PD on the signalling pathways were analysed by western blotting. p‑PD‑treated cells exhibited an upregulated phospho‑stress‑activated protein kinase/JNK protein expression level and downregulated Ras and Raf protein expression levels; however, Akt, Bcl‑2, Bcl‑XL and Bad protein expression levels were not significantly altered compared with the control. In conclusion, p‑PD induced apoptosis by a PTK/Ras/Raf/JNK‑dependent pathway and was independent of the PI3K/Akt pathway in NRK‑52E cells.
对苯二胺（p-PD）是一种潜在致癌物，广泛用于市售染发剂配方中。在本研究中，研究了蛋白酪氨酸激酶（PTK）/Ras/Raf/c-Jun氨基末端激酶（JNK）和磷酸肌醇3激酶（PI3k）/蛋白激酶B（Akt）信号通路对NRK-52E细胞生长的作用。结果表明，p-PD以剂量依赖性方式降低细胞活力。通过细胞周期分析和膜联蛋白V-异硫氰酸荧光素结合试验证实细胞因凋亡而死亡。用二乙酸2',7'-二氯荧光素染色后，与对照组相比，处理后的细胞活性氧（ROS）生成显著增加。通过蛋白质印迹分析p-PD对信号通路的影响。p-PD处理的细胞磷酸化应激激活蛋白激酶/JNK蛋白表达水平上调，Ras和Raf蛋白表达水平下调；然而，与对照组相比，Akt、Bcl-2、Bcl-XL和Bad蛋白表达水平没有显著变化。总之，p-PD通过PTK/Ras/Raf/JNK依赖性途径诱导NRK-52E细胞凋亡，且不依赖于PI3K/Akt途径。
