Guizhou Medical University, Medical Imaging College, Guiyang, China.
Eur Rev Med Pharmacol Sci. 2018 Feb;22(3):695-701. doi: 10.26355/eurrev_201802_14295.
Liver cancer is one of the most common digestive system malignant solid tumors. Its incidence and mortality rates keep high, causing serious mental and economic burden. So far, the exact mechanism of liver cancer onset has not been fully elucidated. Metallothionein (MT) widely exists in various types of organisms with highly conserved structure. It contains the short peptide of cysteine and sulfur protein with high affinity to heavy metals, including cadmium, zinc, and copper. MT1M is an important member of MT family that has been verified to participate in regulating hepatocellular carcinoma, thyroid cancer, cervical cancer, and other tumors. However, MT1M expression and mechanism in hepatoma cells have not been fully elucidated.
Hepatoma cell line HepG2 was divided into control and MT1M group. MT1M plasmid was constructed and transfected to MT1M group. Real-time PCR was used to test MT1M expression. MTT assay was applied to detect HepG2 proliferation. Flow cytometry was performed to determine HepG2 apoptosis. Caspase-3 activity was measured. Western blot was used to detect Bcl-2 and Bax protein levels.
MT1M expression significantly increased after MT1M plasmid transfection compared with control (p < 0.05). MT1M group showed inhibited HepG2 proliferation, declined HepG2 apoptosis, enhanced Caspase-3 activity, reduced Bcl-2 protein level, and upregulated Bax protein compared with control (p < 0.05).
MT1M can suppress HepG2 proliferation and induce HepG2 apoptosis through downregulating Bcl-2, upregulating Bax, and enhancing Caspase-3 activity.
肝癌是最常见的消化系统恶性实体肿瘤之一。其发病率和死亡率居高不下，给患者带来严重的精神和经济负担。迄今为止，肝癌发病的确切机制尚未完全阐明。金属硫蛋白（MT）广泛存在于各种生物中，具有高度保守的结构。它含有短肽半胱氨酸和硫蛋白，对包括镉、锌和铜在内的重金属具有高亲和力。MT1M 是 MT 家族的重要成员，已被证实参与调节肝癌、甲状腺癌、宫颈癌等肿瘤。然而，MT1M 在肝癌细胞中的表达和机制尚未完全阐明。
将肝癌细胞系 HepG2 分为对照组和 MT1M 组。构建 MT1M 质粒并转染至 MT1M 组。实时 PCR 用于检测 MT1M 表达。MTT 检测用于检测 HepG2 增殖。流式细胞术用于检测 HepG2 凋亡。检测 Caspase-3 活性。Western blot 用于检测 Bcl-2 和 Bax 蛋白水平。
与对照组相比，MT1M 质粒转染后 MT1M 表达显著增加（p < 0.05）。与对照组相比，MT1M 组 HepG2 增殖受到抑制，凋亡增加，Caspase-3 活性增强，Bcl-2 蛋白水平降低，Bax 蛋白水平升高（p < 0.05）。
MT1M 可以通过下调 Bcl-2、上调 Bax 和增强 Caspase-3 活性来抑制 HepG2 增殖并诱导 HepG2 凋亡。
