基于CXCR12/CXCR4/CXCR7轴研究熊果酸诱导人甲状腺乳头状癌细胞线粒体凋亡的作用机制  

Mechanism of ursolic acid on inducing mitochondrial apoptosis of human papillary thyroid carcinoma cells by regulating CXCR12/CXCR4/CXCR7 axis

作　　者：马丽华 赵爱兵 吴长明 李子安 MA Li-hua;ZHAO Ai-bing;WU Chang-ming;LI Zi-an(Department of Breast Surgery,Tangshan Workers’Hospital,Tangshan 063000,China;Second Department of Gastrointestinal Surgery,Tangshan Workers’Hospital,Tangshan 063000,China;First Department of Gastrointestinal Surgery,Tangshan Workers’Hospital,Tangshan 063000,China)

机构地区：[1]唐山市工人医院乳腺外科,河北唐山063000 [2]唐山市工人医院胃肠外二科,河北唐山063000 [3]唐山市工人医院胃肠外一科,河北唐山063000

出　　处：《中草药》2023年第19期6370-6376,共7页Chinese Traditional and Herbal Drugs

基　　金：河北省卫生健康委员会科研基金资助项目(20180397)。

摘　　要：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、4.0μmol/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(0.8,1.6,3.2,3.6,4.0μmol/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.

关 键 词：甲状腺乳头状癌 熊果酸 CXCR12/CXCR4/CXCR7轴 线粒体凋亡 增殖 

分 类 号：R285.5[医药卫生—中药学]