Department of Science in Korean Medicine, Kyung Hee University, Seoul, Republic of Korea.
Department of Studies in Molecular Biology, University of Mysore, Manasagangotri, Mysore, Karnataka, India.
J Cell Biochem. 2024 Apr;125(4):e30537. doi: 10.1002/jcb.30537. Epub 2024 Feb 15.
Mitogen-activated protein kinase (MAPK) activation by natural compounds is known to be involved in the induction of apoptosis, paraptosis, and autophagy. Cannabidiol (CBD), a bioactive compound found in Cannabis sativa, is endowed with many pharmacological activities. We investigated the cytotoxic effect of CBD in a panel of colorectal cancer (CRC) cells (HT-29, SW480, HCT-116, and HCT-15). CBD induced significant cytotoxicity as evidenced by the results of MTT assay, live-dead assay, and flow cytometric analysis. Since CBD displayed cytotoxicity against CRC cells, we examined the effect of CBD on apoptosis, paraptosis, and autophagy. CBD decreased the expression of antiapoptotic proteins and increased the Annexin-V-positive as well as TUNEL-positive cells suggesting that CBD induces apoptosis. CBD increased the expression of ATF4 (activating transcription factor 4) and CHOP (CCAAT/enhancer-binding protein homologous protein), elevated endoplasmic reticulum stress, and enhanced reactive oxygen species levels indicating that CBD also promotes paraptosis. CBD also induced the expression of Atg7, phospho-Beclin-1, and LC3 suggesting that CBD also accelerates autophagy. Since, the MAPK pathway is a common cascade that is involved in the regulation of apoptosis, paraptosis, and autophagy, we investigated the effect of CBD on the activation of JNK, p38, and ERK pathways. CBD activated all the forms of MAPK proteins and pharmacological inhibition of these proteins reverted the observed effects. Our findings implied that CBD could induce CRC cell death by activating apoptosis, paraptosis, and autophagy through the activation of the MAPK pathway.
已知天然化合物激活丝裂原活化蛋白激酶 (MAPK) 参与细胞凋亡、副凋亡和自噬的诱导。大麻素（CBD）是大麻中发现的一种生物活性化合物，具有许多药理活性。我们研究了 CBD 在一系列结直肠癌 (CRC) 细胞（HT-29、SW480、HCT-116 和 HCT-15）中的细胞毒性作用。MTT 检测、死活检测和流式细胞术分析结果表明，CBD 诱导了显著的细胞毒性。由于 CBD 对 CRC 细胞显示出细胞毒性，我们检查了 CBD 对细胞凋亡、副凋亡和自噬的影响。CBD 降低了抗凋亡蛋白的表达，并增加了 Annexin-V 阳性和 TUNEL 阳性细胞，表明 CBD 诱导细胞凋亡。CBD 增加了 ATF4（激活转录因子 4）和 CHOP（CCAAT/增强子结合蛋白同源蛋白）的表达，升高了内质网应激水平并增强了活性氧水平，表明 CBD 也促进了副凋亡。CBD 还诱导了 Atg7、磷酸化 Beclin-1 和 LC3 的表达，表明 CBD 也加速了自噬。由于 MAPK 途径是参与细胞凋亡、副凋亡和自噬调节的共同级联，我们研究了 CBD 对 JNK、p38 和 ERK 途径激活的影响。CBD 激活了所有形式的 MAPK 蛋白，并且这些蛋白的药理学抑制逆转了观察到的效应。我们的研究结果表明，CBD 通过激活 MAPK 途径诱导 CRC 细胞死亡，从而诱导细胞凋亡、副凋亡和自噬。
