Department of Thyroid, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, China.
Department of Nuclear Medicine, The Affiliated Yantai Yuhuangding Hospital of Qingdao University, Yantai 264000, China.
Steroids. 2020 Jul;159:108585. doi: 10.1016/j.steroids.2020.108585. Epub 2020 Jan 23.
To explore the effects of 1,25-dihydroxy vitamin D3 [1,25-(OH)D] on the proliferation and apoptosis of papillary thyroid carcinoma and to investigate its possible mechanism.
The papillary thyroid carcinoma cell line TPC-1 was cultured, and the cells were divided into control group, the 1,25-(OH)D group, and the 1,25-(OH)D + ML-098 (Ras agonist) group. Cell proliferation was observed by MTT. The colony formation viability of cells was detected by the plate cloning assay. Cell migration was observed by the scratch assay. Apoptosis was detected by flow cytometry. The expression of Ki67 and Caspase-3, and the activity of Ras-MEK-ERK pathway were detected by western blot.
Compared with the Control group, the proliferation, colony formation and migration ability of cells in the drug group were significantly decreased. The number of apoptotic cells was significantly increased, the expression of Ki67 protein was decreased, and the expression of Caspase-3 protein was upregulated. The phosphorylation levels of Ras, p-ERK1/2, and p-MEK were decreased. Compared with the drug group, the cloning and migration biological activity of cells in the 1,25-(OH)D + ML-098 group was significantly enhanced (p < 0.05). The number of apoptotic cells was decreased, while the Ki67 protein level was increased. In addition, the Caspase-3 protein level was decreased, and the Ras-MEK-ERK level was also enhanced. Furthermore, the antitumor activity of 1,25-(OH)D was reversed by the Ras agonist ML-098.
1,25-(OH)D can inhibit the activity and promote apoptosis of the papillary thyroid carcinoma cell line TPC-1, and its mechanism may be related to the inhibition of the Ras-MEK-ERK pathway activity, thus affecting the proliferation and expression of apoptosis-related proteins.
探讨 1,25-二羟维生素 D3[1,25-(OH)D]对甲状腺乳头状癌细胞增殖和凋亡的影响，并探讨其可能的机制。
培养甲状腺乳头状癌细胞系 TPC-1，将细胞分为对照组、1,25-(OH)D 组和 1,25-(OH)D+ML-098（Ras 激动剂）组。MTT 观察细胞增殖情况，平板克隆实验检测细胞集落形成活力，划痕实验观察细胞迁移能力，流式细胞术检测细胞凋亡情况，Western blot 检测 Ki67 和 Caspase-3 表达及 Ras-MEK-ERK 通路活性。
与对照组相比，药物组细胞增殖、集落形成和迁移能力明显降低，凋亡细胞数量明显增加，Ki67 蛋白表达减少，Caspase-3 蛋白表达上调，Ras、p-ERK1/2 和 p-MEK 磷酸化水平降低。与药物组相比，1,25-(OH)D+ML-098 组细胞的克隆和迁移生物活性明显增强（p<0.05），凋亡细胞数量减少，Ki67 蛋白水平升高，Caspase-3 蛋白水平降低，Ras-MEK-ERK 水平也升高。此外，Ras 激动剂 ML-098 逆转了 1,25-(OH)D 的抗肿瘤活性。
1,25-(OH)D 可抑制甲状腺乳头状癌细胞系 TPC-1 的活性并促进其凋亡，其机制可能与抑制 Ras-MEK-ERK 通路活性有关，从而影响增殖和凋亡相关蛋白的表达。
