Sanaei Masumeh, Kavoosi Fraidoon
Research Center for Non-Communicable Diseases, Jahrom University of Medical Sciences, Jahrom, Iran.
Gastroenterol Hepatol Bed Bench. 2021 Fall;14(4):323-333.
The current study investigated the effect of trichostatin A (TSA) on mitochondrial/intrinsic [pro- (Bax, Bak, and Bim) and anti- (Bcl-2, Bcl-xL, and Mcl-1) apoptotic genes] and cytoplasmic/extrinsic (DR4, DR5, FAS, FAS-L, and TRAIL genes) pathways, histone deacetylase 1, 2, and 3, p53, p73, cell viability, and apoptosis in hepatocellular carcinoma (HCC) HCCLM3, MHCC97H, and MHCC97L cell lines.
Modulation of the acetylation status of histones, histones modification, plays an important role in regulating gene transcription and expression. Histone deacetylation controlled by histone deacetylases (HDACs) leads to gene downregulation. Histone deacetylase inhibitors (HDACIs) are an emerging class of therapeutics with potential anticancer effects. They can induce apoptosis by activating both extrinsic and intrinsic apoptotic pathways.
HCCLM3, MHCC97H, and MHCC97L cells were cultured and treated with TSA. To determine viability, apoptosis, and the relative expression level of the mentioned genes, MTT assay, cell apoptosis assay, and qRT-PCR, respectively, were conducted.
TSA up-regulated Bax, Bak, Bim, DR4, DR5, FAS, FAS-L, TRAIL, p53, and p73 and down-regulated Bcl-2, Bcl-xL, Mcl-1, histone deacetylases 1, 2, and 3 significantly, resulting in apoptosis induction. Maximal and minimal apoptosis was seen in the MHCC97H and HCCLM3 cell lines (93.94% and 39.68%, respectively) after 24 and 48 h. Therefore, the MHCC97H cell line was more sensitive to TSA.
The current findings demonstrated that the HDAC inhibitor TSA can induce apoptosis and inhibit cell growth through both mitochondrial/intrinsic and cytoplasmic/extrinsic apoptotic pathways in hepatocellular carcinoma HCCLM3, MHCC97H, and MHCC97L cell lines.
本研究调查了曲古抑菌素A（TSA）对线粒体/内在途径（促凋亡基因Bax、Bak和Bim以及抗凋亡基因Bcl-2、Bcl-xL和Mcl-1）和细胞质/外在途径（DR4、DR5、FAS、FAS-L和TRAIL基因）、组蛋白去乙酰化酶1、2和3、p53、p73、细胞活力以及肝癌（HCC）HCCLM3、MHCC97H和MHCC97L细胞系凋亡的影响。
组蛋白乙酰化状态的调节，即组蛋白修饰，在调节基因转录和表达中起重要作用。由组蛋白去乙酰化酶（HDACs）控制的组蛋白去乙酰化导致基因下调。组蛋白去乙酰化酶抑制剂（HDACIs）是一类新兴的具有潜在抗癌作用的治疗药物。它们可通过激活外在和内在凋亡途径诱导细胞凋亡。
培养HCCLM3、MHCC97H和MHCC97L细胞并用TSA处理。分别通过MTT法、细胞凋亡检测和qRT-PCR来确定细胞活力、凋亡情况以及上述基因的相对表达水平。
TSA显著上调Bax、Bak、Bim、DR4、DR5、FAS、FAS-L、TRAIL、p53和p73，同时显著下调Bcl-2、Bcl-xL、Mcl-1、组蛋白去乙酰化酶1、2和3，从而诱导细胞凋亡。在24小时和48小时后，MHCC97H和HCCLM3细胞系分别出现最大和最小凋亡率（分别为93.94%和39.68%）。因此，MHCC97H细胞系对TSA更敏感。
目前的研究结果表明，HDAC抑制剂TSA可通过线粒体/内在和细胞质/外在凋亡途径诱导肝癌HCCLM3、MHCC97H和MHCC97L细胞系凋亡并抑制细胞生长。
