夏枯草诱导人甲状腺乳头状癌细胞K1增殖和凋亡的影响及其作用机制

目的:探索夏枯草对人甲状腺乳头状癌细胞(K1)增殖和诱导K1细胞凋亡的影响及其可能的作用机制。方法:采用MTT比色法测定2~200 mg/mL夏枯草作用24 h对K1细胞增殖的影响;采用Hoechst染色法和流式细胞术(Annexin V-FITC/PI联合标记)观察0.3、0.6、1.2、2.4、4.8 mg/mL夏枯草作用24 h K1细胞凋亡的影响;采用蛋白质免疫印迹法(Western blotting)测定1.2、2.4、4.8 mg/mL夏枯草作用24h对K1细胞凋亡相关蛋白表达的影响。结果:在2~200 mg/mL的浓度范围内夏枯草作用24 h对K1细胞增殖有明显抑制作用(P0.05),IC50值为2.427 mg/mL。流式细胞术检测结果显示夏枯草可诱导K1细胞凋亡,2.4、4.8 mg/mL组K1细胞总凋亡率分别为(11.35±0.92)%、(44.57±3.07)%,与对照组相比明显升高(P0.05);与对照组相比,1.2、2.4、4.8 mg/mL夏枯草作用24 h胞浆内凋亡蛋白Bax、细胞色素C(Cyto C)、Caspase-9、活化的Caspase-3(Cleaved Caspase-3)表达增多。结论:夏枯草能抑制人甲状腺癌K1细胞增殖并诱导其凋亡,其机制可能与活化线粒体凋亡通路有关。

Effects and Mechanism of Prunella Vulgaris L. on the Proliferation and Apoptosis of Human Papillary Thyroid Cancer K1 Cells

ABSTRACT Objective: To explore the effect and the possible mechanism of Prunella vulgaris L. on the cell proliferation and apop- tosis in the human Papillary thyroid cancer K1 cells. Methods: The inhibition ratio of Prunella vulgaris L. on the human Papillary thyroid cancer cells K1 was tested by MTT assay, the influence of different concentrations of Prunella vulgaris L. on the apoptosis of human Pap- illary thyroid cancer K1 cells was observed by Hoechst staining method; the apoptotic rates were examined by flow cytometry analysis. The expression of apoptotic protein was detected by Western blotting. Results: Prunella vulgaris L. could obviously inhibit the prolifera- tion of K1 cells in 2~200 mg/mL concentration range(P<0.05). The IC₅₀ with 24 h of Prunella vulgaris L. on the K1 cell was 2.427 mg/mL. Flow cytometry analysis showed that: 2.4, 4.8 mg/mL Prunella vulgaris L. could induce the apoptosis of K1 cell, the apoptotic rates were (11.35±0.92)% and (44.57±3.07)% respectively. Western blotting showed that the expressions of apoptotic protein Bcl-2, Caspase-9, Caspase-3 of Prunella vulgaris L. were decreased and the expressions of Cleaved Caspase 3, Caspase-9, CytoC, Bax were in- creased compared with the control group. Conclusion: Prunella vulgaris L. could inhibit the cell proliferation and induce the apoptosis of human papillary thyroid cancer K1 cells, which might be related to the activation of mitochondrial pathway.