Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea.
Laboratory of Biochemistry and Immunology, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 28644, Republic of Korea.
Phytomedicine. 2018 Jan 15;39:100-110. doi: 10.1016/j.phymed.2017.12.006. Epub 2017 Dec 7.
Phytochemicals are derived from plants, vegetables and daily products and exert chemopreventive effects. Malignant melanoma is highly metastatic, and melanoma patients can develop chemotherapeutic resistance against conventional melanoma therapies.
In the present study, we investigated the anti-cancer effect of the phytochemicals kaempferol (Kaem), genistein (Gen), and 3'3-diindolylmethane (DIM) on melanoma cell viability. We also evaluated the altered expression of cell cycle-related genes. We verified the production of intracellular reactive oxygen species (ROS) and endoplasmic reticulum (ER) stress at both the protein and cellular level using a western blot, TUNEL assay, and Dihydrodichlorofluorescein diacetate (DCF-DA) assay.
Treatment of A375SM melanoma cells with phytochemicals resulted in inhibition of cell growth. Treatment with phytochemicals increased the gene expression of p21 and decreased the gene expression of cyclin E and/or cyclin B. The three phytochemicals activated the ROS-p38-p53 apoptotic pathway by increasing the level of phosphorylated p38 MAPK and p53, and they activated the ER stress-mediated apoptotic pathway by increasing the level of phosphorylated eIF2α and C/EBP homologous protein (CHOP). Both the ROS-p38-p53 and ER stress-mediated pathway induced the mitochondrial apoptotic pathway by attenuating Bcl-2 expression and upregulating BAX. Detection of morphological changes demonstrated that Kaem and Gen can induce differentiation in A375SM cell line.
These results indicate that phytochemicals are potentially useful in treatments for melanoma due to their ability to inhibit melanoma cell growth and division via the ROS and ER stress pathway.
植物化学物质来源于植物、蔬菜和日常产品，具有化学预防作用。恶性黑色素瘤具有高度转移性，黑色素瘤患者可能对常规黑色素瘤治疗产生化疗耐药性。
在本研究中，我们研究了植物化学物质山奈酚（Kaem）、染料木黄酮（Gen）和 3'3-二吲哚基甲烷（DIM）对黑色素瘤细胞活力的抗癌作用。我们还评估了细胞周期相关基因的改变表达。我们使用 Western blot、TUNEL 测定和二氢二氯荧光素二乙酸酯（DCF-DA）测定在蛋白质和细胞水平上验证了细胞内活性氧（ROS）和内质网（ER）应激的产生。
植物化学物质处理 A375SM 黑色素瘤细胞导致细胞生长抑制。植物化学物质处理增加了 p21 的基因表达，降低了细胞周期蛋白 E 和/或细胞周期蛋白 B 的基因表达。三种植物化学物质通过增加磷酸化 p38 MAPK 和 p53 的水平激活 ROS-p38-p53 凋亡途径，并通过增加磷酸化 eIF2α 和 C/EBP 同源蛋白（CHOP）的水平激活 ER 应激介导的凋亡途径。ROS-p38-p53 和 ER 应激介导的途径通过减弱 Bcl-2 表达和上调 BAX 诱导线粒体凋亡途径。形态变化的检测表明 Kaem 和 Gen 可以诱导 A375SM 细胞系分化。
这些结果表明，由于植物化学物质能够通过 ROS 和 ER 应激途径抑制黑色素瘤细胞生长和分裂，因此它们在黑色素瘤治疗中具有潜在的用途。
