目的 考察熊果酸对人甲状腺乳头状癌TPC-1细胞凋亡的影响及其作用机制。方法 TPC-1细胞分别加入不同浓度（0.8、1.6、3.2、3.6、4.0 μmol/L）的熊果酸处理，采用CCK-8试剂盒检测细胞增殖；采用克隆形成实验检测细胞克隆形成能力；采用流式细胞术检测细胞凋亡率；采用Fura-3/AM、JC-1探针分别检测细胞内Ca²⁺水平和线粒体膜电位；采用Western blotting和PCR检测线粒体凋亡相关蛋白和趋化因子受体12（C-X-C chemokine receptor type 12，CXCR12）/CXCR4/CXCR7轴相关基因表达。结果 熊果酸显著降低TPC-1细胞增殖和克隆形成能力（P<0.01），显著降低线粒体膜电位（P<0.05、0.01），显著升高细胞内Ca²⁺水平和细胞凋亡率（P<0.01），显著下调B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl-2）蛋白表达和CXCR12/CXCR4/CXCR7轴相关基因表达（P<0.01），显著上调细胞色素C（cytochrome-C，Cyt-C）、Bcl-2相关X蛋白（Bcl-2 associated X protein，Bax）、半胱氨酸天冬氨酸蛋白酶-9（cystein-asparate protease-9，Caspase-9）和Caspase-3蛋白表达（P<0.05、0.01）。结论 熊果酸可能通过抑制CXCR12/CXCR4/CXCR7轴活化进而诱导TPC-1细胞线粒体凋亡，从而发挥抗甲状腺乳头状癌的作用。

Objective To investigate the effect and mechanism of ursolic acid on apoptosis of human papillary thyroid carcinoma TPC-1 cells. Methods TPC-1 cells were treated with different concentrations (0.8, 1.6, 3.2, 3.6, 4.0 μmol/L) of ursolic acid, and the cell proliferation was detected by CCK-8 kit. The ability of cell clone formation was detected by clone formation experiment. Apoptosis rate was detected by flow cytometry. Fura-3/AM and JC-1 probes were used to detect intracellular Ca²⁺ level and mitochondrial membrane potential respectively. Western blotting and PCR were used to detect the expressions of mitochondrial apoptosis-related proteins and chemokine receptor type 12 (CXCR12)/CXCR4/CXCR7 axis-related genes. Results Ursolic acid significantly reduced the proliferation and clone formation ability of TPC-1 cells (P < 0.01), significantly reduced mitochondrial membrane potential (P < 0.05, 0.01), significantly increased the intracellular Ca²⁺ level and apoptosis rate (P < 0.01), significantly down-regulated expressions of B-cell lymphoma-2 (Bcl-2) protein and CXCR12/CXCR4/CXCR7 axis-related genes (P < 0.01), significantly up-regulated expressions of cytochrome-C (Cyt-C), Bcl-2 associated X protein (Bax), cysteine-aspartic protease-9 (Caspase-9) and Caspase-3 proteins (P < 0.05, 0.01). Conclusion Ursolic acid may induce mitochondrial apoptosis in TPC-1 cells by inhibiting the activation of CXCR12/CXCR4/CXCR7 axis, thus playing an anti-thyroid papillary carcinoma role.

R285.5

河北省卫生健康委员会科研基金资助项目（20180397）