中华猕猴桃根氯仿部分抑制喉癌HEP-2细胞增殖作用的研究

Study on Inhibition Effect of Actinidia Chinensis Planch Extracted by Chloroform Proliferation of HEP-2 Cells

目的:探讨中华猕猴桃根氯仿提取部分(Actinidiachinensis planch extracted by chloroform,EC-ACP)对喉癌HEP-2的增殖抑制及诱导凋亡的作用,并探讨可能的机制。方法:运用CCK-8法检测EC-ACP对喉癌HEP-2的抑制作用及IC50值;流式细胞术检测EC-ACP对喉癌HEP-2的凋亡率及周期的影响;Western blot法检测EC-ACP对喉癌HEP-2细胞蛋白表达的影响。结果:24、48、36 h后不同浓度的EC-ACP作用于HEP-2细胞均能明显的抑制细胞增殖(P<0.05),且存在时间-浓度依赖性。Annexin V-FITC/PI双染法结果提示EC-ACP作用于喉癌HEP-2细胞48 h后,随着浓度的增加,凋亡率逐渐增加。PI染色结果提示EC-ACP将HEP-2细胞周期阻滞于G1期。Western-blot结果显示随着加药浓度的增加,周期相关蛋白P53表达逐渐增大,周期相关蛋白Cyclin D1表达减少。结论:中华猕猴桃根氯仿提取部分可以抑制HEP-2细胞的增值,其发生可能与其促进p53,抑制cyclin D1基因表达有关。

Objective ： To study the inhibition of Actinidia chinensis planch extracted by chloroform on proliferation of human cancer cell line HEP - 2 cells and induction on apoptosis and explore its mechanism. Method： Use the CCK - 8 assay, flow cytometric （FCM） analysis and Western blot method to detect the EC -ACP＇s affect on HEP -2 cells. Result： The result of CCK - 8 showed that after EC - ACP traetment, the proliferation of HEP - 2 cell was obviously inhibited in a dose - and time -dependent manner. The apoptotic cells were detected after treating with 40 μg/mL,80 μg/mL and 160 μg/mL of the EC - ACP for 48 h and the effect was enhanced as the dosage increase. The percentages of the G0/G1 phase cells were increased and that of S phase cells were decreased after treated by EC - ACP for 48 h. Apoptptic protein P53 was up - regulated, while cyclinD1 protein down - regulated. Conclusion ： EC - ACP inhibits the proliferation of HEP - 2 cells, probably through the up - regulation of P53 and down - regulation of cyclinD1.