Mohammadi Saeed, Seyedhosseini Fakhri Sadat, Behnampour Nasser, Yazdani Yaghoub
a Student Research Committee, Department of Molecular Medicine, School of Advanced Technologies in Medicine , Golestan University of Medical Sciences , Gorgan , Iran.
b Infectious Diseases Research Center and Laboratory Science Research Center , Golestan University of Medical Sciences , Gorgan , Iran.
J Recept Signal Transduct Res. 2017 Oct;37(5):506-514. doi: 10.1080/10799893.2017.1360351.
The role of aryl hydrocarbon receptor (AhR) in carcinogenesis has been studied recently. Indole-3-carbinol (I3C) is an AhR agonist and a potential anticancer agent. Here, we investigated the effects of I3C on cell cycle progression and apoptosis through activation of AhR on THP-1 acute myeloid leukemia (AML) cell line.
MTT viability assay was used to measure the cytotoxic effects of I3C on THP-1 cells. Apoptosis and cell cycle assays were investigated using flow cytometry. Real time RT-PCR was conducted to measure the alterations in the expression of AhR gene, key genes associated with AhR activation (IL1β and CYP1A1) and major genes involved in cell cycle regulation and apoptosis including P27, P21, CDK2, P53, BCL2 and FasR.
Our findings revealed that I3C inhibits the proliferation of THP-1 cells in a dose- and time-dependent manner with minimal toxicity over normal monocytes. The AhR target genes (CYP1A1, IL1β) were overexpressed upon I3C treatment (p < .05 to p < .001). The antiproliferative effects of I3C were in association with programed cell death. I3C downregulated BCL2 and upregulated FasR in THP-1 cells (p < .05 to p < .001). G1 cell cycle arrest was also observed using flow cytometry. G1-acting cell cycle genes (P21, P27 and P53) were overexpressed (p < .05 to p < .001), while CDK2 was downregulated upon I3C treatment (p < .01 to p < .001).
I3C could exert its antileukemic effects through AhR activation which is associated with programed cell death and G1 cell cycle arrest in a dose- and time-dependent manner. Therefore, AhR could be targeted as a novel treatment possibility in AML.
近期对芳烃受体（AhR）在致癌过程中的作用进行了研究。吲哚 - 3 - 甲醇（I3C）是一种AhR激动剂和潜在的抗癌剂。在此，我们通过激活AhR，研究了I3C对THP - 1急性髓系白血病（AML）细胞系细胞周期进程和细胞凋亡的影响。
采用MTT活力测定法检测I3C对THP - 1细胞的细胞毒性作用。使用流式细胞术研究细胞凋亡和细胞周期检测。进行实时RT - PCR以测量AhR基因、与AhR激活相关的关键基因（IL1β和CYP1A1）以及参与细胞周期调控和细胞凋亡的主要基因（包括P27、P21、CDK2、P53、BCL2和FasR）表达的变化。
我们的研究结果显示，I3C以剂量和时间依赖性方式抑制THP - 1细胞的增殖，对正常单核细胞的毒性最小。I3C处理后，AhR靶基因（CYP1A1、IL1β）过表达（p < 0.05至p < 0.001）。I3C的抗增殖作用与程序性细胞死亡相关。I3C下调THP - 1细胞中的BCL2并上调FasR（p < 0.05至p < 0.001）。使用流式细胞术还观察到G1期细胞周期阻滞。G1期作用的细胞周期基因（P21、P27和P53）过表达（p < 0.05至p < 0.001），而I3C处理后CDK2下调（p < 0.01至p < 0.001）。
I3C可通过激活AhR发挥其抗白血病作用，这与程序性细胞死亡和G1期细胞周期阻滞呈剂量和时间依赖性相关。因此，AhR可作为AML的一种新的治疗靶点。
