Sanaei Masumeh, Kavoosi Fraidoon
Department of Anatomy, Research Center for Non Communicable Diseases, Jahrom University of Medical Sciences, Jahrom, Iran.
Int J Prev Med. 2023 May 27;14:63. doi: 10.4103/ijpvm.ijpvm_191_21. eCollection 2023.
The alteration of DNA cytosine methylation is one of the most common epigenetic changes that can play a significant role in human cancers. The enzymes involved in DNA methylation of promoter regions of the genes are DNA methyltransferases (DNMTs). The therapeutic activities and apoptotic effects of DNA methyltransferase inhibitors (DNMTIs) have been reported in various cancers. This study was assigned to assess the effect of zebularine on intrinsic and extrinsic pathways, DNAT 1, 3a, and 3b, p21, and p53, viability, and apoptosis in hepatocellular carcinoma (HCC) cell lines.
Hepatocellular carcinoma cell lines (HCCLM3, MHCC97H, and MHCC97L) were purchased from the National Cell Bank of Iran, Pasteur Institute, treated with zebularine, and the MTT assay was performed. Then, flow cytometry assay and real-time RT-PCR analysis were performed with zebularine. Statistical comparisons between groups were made using GraphPad Prism software version 8.0. A significant difference was considered as < 0.05.
Zebularine up-regulated DR4, DR5, FAS, FAS-L, TRAIL, Bax, Bak, Bim, p21WAF/CIP1 (p21), and p53 and down-regulated DNMTs (DNAT 1, 3a, and 3b), Bcl-2, Bcl-xL, and Mcl-1, significantly resulting in apoptosis induction in HCC cell lines. Maximal and minimal apoptosis was seen in HCCLM3 and MHCC97L cell lines, respectively.
Our findings indicated that DNMTI zebularine can induce apoptosis and inhibit cell growth through both pathways (extrinsic and intrinsic) in HCC cell lines HCCLM3, MHCC97H, and MHCC97L.
DNA胞嘧啶甲基化改变是最常见的表观遗传变化之一，可在人类癌症中发挥重要作用。参与基因启动子区域DNA甲基化的酶是DNA甲基转移酶（DNMTs）。DNA甲基转移酶抑制剂（DNMTIs）的治疗活性和凋亡作用已在多种癌症中得到报道。本研究旨在评估zebularine对肝细胞癌（HCC）细胞系内在和外在途径、DNAT 1、3a和3b、p21和p53、活力及凋亡的影响。
从伊朗巴斯德研究所国家细胞库购买肝细胞癌细胞系（HCCLM3、MHCC97H和MHCC97L），用zebularine处理，然后进行MTT试验。接着，对zebularine进行流式细胞术检测和实时RT-PCR分析。使用GraphPad Prism软件8.0版本进行组间统计比较。差异有统计学意义定义为P<0.05。
zebularine上调DR4、DR5、FAS、FAS-L、TRAIL、Bax、Bak、Bim、p21WAF/CIP1（p21）和p53，下调DNMTs（DNAT 1、3a和3b）、Bcl-2、Bcl-xL和Mcl-1，显著导致HCC细胞系凋亡诱导。分别在HCCLM3和MHCC97L细胞系中观察到最大和最小凋亡。
我们的研究结果表明，DNMTI zebularine可通过外在和内在两条途径诱导HCC细胞系HCCLM3、MHCC97H和MHCC97L凋亡并抑制细胞生长。
