白藜芦醇对人甲状腺乳头状癌IHH4细胞增殖和凋亡的影响

目的:观察白藜芦醇对人甲状腺乳头状癌细胞(IHH4)增殖和凋亡的影响,并初步探讨其可能的机制?方法:采用CCK-8法检测白藜芦醇对IHH4细胞增殖的影响;倒置显微镜?透射电镜?DAPI染色观察细胞的形态学变化;细胞免疫荧光和Western blot法检测细胞内cleaved caspase-3蛋白的表达;应用流式细胞技术(flow cytometry,FCM)观察细胞周期和凋亡的改变?结果:①白藜芦醇可以呈时间和浓度依赖性抑制IHH4细胞增殖;②与正常对照相比,加入白藜芦醇后IHH4细胞皱缩?变圆?脱落,细胞质中出现大小不等的空泡,胞质内容物增多;透射电镜下细胞核的染色质高度凝聚?边缘化;DAPI染色后荧光显微镜下观察,细胞核呈致密浓染;③细胞免疫荧光和Western blot显示,随着白藜芦醇作用浓度和时间的增加cleaved caspase-3蛋白的表达明显增加;④流式Annexin V-FITC/PI双染检测显示白藜芦醇呈浓度依赖性诱导IHH4细胞凋亡?白藜芦醇可引起细胞S期阻滞?结论:白藜芦醇可抑制人甲状腺乳头状癌IHH4细胞增殖,诱导IHH4细胞凋亡?白藜芦醇阻滞细胞于S期,可能是抑制IHH4细胞增殖?促使其凋亡的原因之一?

Effects of resveratrol on proliferation and apoptosis of human papillary thyroid carcinoma IHH4 cell

Objective:To investigate the effects of resveratrol on proliferation and apoptosis of human papillary thyroid carcinoma IHH4 cells and its possible mechanisms. Methods:The inhibition rate of resveratrol on IHH4 cells proliferation was detected by CCK-8 assay. Morphological changes of IHH4 cells were observed by inverted microscope,transmission electron microscope and DAPI staining,respectively. The expression of cleaved caspase-3 was detected by immunofluorescence cytochemistry and Western blot. The change of cell apoptosis and cell cycle distribution were analyzed by flow cytometry. Results:Resveratrol inhibited the growth of IHH4 cells in a time- and dose-dependent manner. Compared with normal control,after resveratrol treatment,IHH4 cells shrank,turned round and began to shed off,various sizes of vacuoles appeared in the cytoplasm,intracytoplasmic content increased under optical microscope. The transmission electron microscope examination showed pyknosis and margination of apoptotic cell nucleus chromatin. DAPI staining showed that the normal nucleichromatin was relatively dispersed with regular shape and smooth surface,while cells treated with resveratrol displayed condensed and fragmented nuclei,which were typical features of apoptosis. Immunofluorescence cytochemistry and Western blot revealed that the expression of cleaved caspase-3 increased significantly in a time- and dose-dependent manner. Resveratrol induced apoptosis of IHH4 cells in a dose-dependent manner with S phase cycle arrest. Conclusion:Resveratrol could inhibit proliferation and induce apoptosis of human IHH4 cells in vitro. One of the possible mechanisms is related to S phase cycle arrest.