Department of Pediatric Surgery, The Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Stem Cell Biology and Therapy Laboratory, Ministry of Education Key Laboratory of Child Development and Disorders, Chongqing Stem Cell Therapy Engineering Technical Center, The Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Int J Oncol. 2016 Jan;48(1):349-57. doi: 10.3892/ijo.2015.3235. Epub 2015 Nov 6.
Hepatocellular carcinoma (HCC) has the characristics of tumor invasiveness, frequent intrahepatic spread and extra hepatic metastases, which affects the therapy efficiency and prognosis. Epithelial-mesenchymal transition (EMT) is now recognized as a key process in tumor invasion, metastasis and the generation of cancer initiating cells. All-trans retinoic acid (ATRA) is currently used as a potential chemo-therapeutic or chemo-preventive agent because of its anti-proliferative, pro-apoptotic and antioxidant properties. This study investigated the effects of ATRA at different concentrations on the proliferation, migration, invasion, differentiation and functions of the mouse hepa1-6 hepatocarcinoma cell line and explored whether ATRA regulates EMT in the antitumor process. Trypan blue staining and colony formation assay were used to detect cell proliferation. Wound-healing assay and Transwell Matrigel assay were performed to examine migration. Invasion was assessed by using Transwell invasion assay. In the present study, ATRA significantly inhibited the cell growth, colony formation, migration, and invasion capability of hepa1-6 cells in a dose-dependent manner. Furthermore, ATRA at low concentration (0.1 µmol/l) could generate these influences. After treated in the ATRA medium, the expression of mature hepatic markers ALB (albumin), CK18 (cytokeratin 18), TAT (tyrosine aminotransferase), ApoB (apolipoprotein B) decreased and that of hepatocarcinoma marker AFP (α fetoprotein) increased. At day 7 after ATRA induction, hepa1-6 cells showed comparable indocyanine green (ICG) uptake and glycogen storage function to the blank control. The mRNA expression of mesenchymal markers N-cadherin, vimentin, snail and twist decreased, while expression of epithelial marker E-cadherin increased in hepa1-6 cells after treated with ATRA. Therefore, this study demonstrates that ATRA remarkably suppressed the proliferation, migration, invasion of hepa1-6 hepatocarcinoma cell line and effectively induced its differentiation and liver functions in vitro through the reversal of EMT. HCC may be more sensitive to ATRA than other cancers, suggesting the prospective usefulness of ATRA in the treatment of HCC.
肝细胞癌 (HCC) 具有肿瘤侵袭性、频繁的肝内扩散和肝外转移的特点，这影响了治疗效果和预后。上皮-间充质转化 (EMT) 现在被认为是肿瘤侵袭、转移和癌症起始细胞产生的关键过程。全反式视黄酸 (ATRA) 因其具有抗增殖、促凋亡和抗氧化特性，目前被用作潜在的化疗或化学预防药物。本研究探讨了不同浓度的 ATRA 对小鼠 hepa1-6 肝癌细胞系增殖、迁移、侵袭、分化和功能的影响，并探讨了 ATRA 是否在抗肿瘤过程中调节 EMT。台盼蓝染色和集落形成实验用于检测细胞增殖。划痕愈合实验和 Transwell Matrigel 实验用于检测迁移。Transwell 侵袭实验用于评估侵袭。在本研究中，ATRA 显著抑制了 Hepa1-6 细胞的生长、集落形成、迁移和侵袭能力，呈剂量依赖性。此外，低浓度 (0.1μmol/l) 的 ATRA 也能产生这些影响。在 ATRA 培养基中处理后，成熟肝标志物 ALB（白蛋白）、CK18（细胞角蛋白 18）、TAT（酪氨酸转氨酶）、ApoB（载脂蛋白 B）的表达降低，肝癌标志物 AFP（α 胎蛋白）的表达增加。在 ATRA 诱导后第 7 天，Hepa1-6 细胞的吲哚菁绿 (ICG) 摄取和糖原储存功能与空白对照组相当。ATRA 处理后 Hepa1-6 细胞中间充质标志物 N-钙粘蛋白、波形蛋白、snail 和 twist 的 mRNA 表达降低，上皮标志物 E-钙粘蛋白的表达增加。因此，本研究表明，ATRA 可显著抑制 Hepa1-6 肝癌细胞系的增殖、迁移和侵袭，并通过 EMT 的逆转有效诱导其分化和肝功能。HCC 可能比其他癌症对 ATRA 更敏感，这表明 ATRA 在 HCC 治疗中的潜在应用价值。
