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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。
Objective: To observe the effects of resveratrol on the proliferation and apoptosis of human thyroid papillary carcinoma cells (IHH4) and to explore its possible mechanism. Methods: The effect of resveratrol on the proliferation of IHH4 cells was detected by CCK-8 method. Morphological changes of cells were observed by inverted microscope, transmission electron microscope and DAPI staining. The expression of cleaved caspase-3 protein was detected by immunofluorescence and Western blot The changes of cell cycle and apoptosis were observed by flow cytometry (FCM). Results: 1 Resveratrol could inhibit the proliferation of IHH4 cells in a time and concentration-dependent manner. 2 Compared with the normal control group, resveratrol reduced the size of IHH4 cells in the cytoplasm, The contents of vacuoles and cytoplasm increased; the chromatin of the nucleus under the TEM was highly condensed and marginalized; DAPI staining showed that the nuclei were densely stained under the fluorescence microscope; immunofluorescence and Western blot showed that with the resveratrol The concentration and time of cleaved caspase-3 protein expression increased significantly; 4-flow Annexin V-FITC / PI double staining showed resveratrol concentration-dependently induced IHH4 cell apoptosis. Resveratrol can cause cell S phase arrest. Conclusion: Resveratrol can inhibit the proliferation of human thyroid papillary carcinoma IHH4 cells and induce the apoptosis of IHH4 cells. Resveratrol blocks cells in S phase, which may be one of the reasons that inhibits IHH4 cell proliferation and induces apoptosis.