本文選題：芹菜素 + 膠質(zhì)瘤　； 參考：《江蘇大學(xué)》2017年碩士論文

【摘要】：目的探討不同濃度芹菜素對膠質(zhì)瘤SHG-44細(xì)胞增殖抑制和誘導(dǎo)凋亡的作用。方法我們分別以不同濃度(20、40、80μmol/L)的芹菜素處理SHG-44膠質(zhì)瘤細(xì)胞24、48、72 h,采用MTT法檢測不同濃度芹菜素對膠質(zhì)瘤細(xì)胞增殖抑制率的影響。運(yùn)用PI單染流式細(xì)胞儀檢測法觀察芹菜素處理對膠質(zhì)瘤24 h后,細(xì)胞周期的分布情況。采用Hoechst 33258、Tunel染色法、AnnexinV-FITC/PI雙染流式細(xì)胞儀檢測技術(shù)探究不同濃度芹菜素對膠質(zhì)瘤細(xì)胞凋亡的影響。通過蛋白印跡方法(Western blot)觀察細(xì)胞周期相關(guān)蛋白CDK6、CDC25A及凋亡途徑相關(guān)蛋白Bax、Bcl-2的表達(dá)量的變化。結(jié)果芹菜素對SHG-44細(xì)胞的增殖抑制作用顯著,且隨藥物作用時(shí)間和作用濃度的增加而增強(qiáng)。細(xì)胞周期檢測顯示芹菜素使膠質(zhì)瘤細(xì)胞發(fā)生G2/M期阻滯。Hoechst33258免疫熒光染色顯示芹菜素處理組膠質(zhì)瘤細(xì)胞可見染色質(zhì)固縮、核斷裂和調(diào)亡小體等典型細(xì)胞凋亡表現(xiàn),TUNEL免疫熒光染色顯示凋亡細(xì)胞出現(xiàn)綠色熒光。流式細(xì)胞儀分析結(jié)果顯示,隨著芹菜素處理濃度的增加,SHG-44細(xì)胞的凋亡率逐漸增高,不同濃度芹菜素作用24 h的結(jié)果顯示該藥物能呈濃度依賴性地誘導(dǎo)SHG-44細(xì)胞凋亡。芹菜素處理組細(xì)胞的CDK6、CDC25A蛋白表達(dá)量逐漸下降,Bax蛋白表達(dá)量逐漸增加,Bcl-2蛋白表達(dá)量逐漸降低,Bax/Bcl-2比值逐漸增大。結(jié)論在體外實(shí)驗(yàn)中,芹菜素可抑制膠質(zhì)瘤SHG-44細(xì)胞增殖,并呈時(shí)間和劑量依賴性。同時(shí)芹菜素可誘導(dǎo)膠質(zhì)瘤SHG-44發(fā)生凋亡;芹菜素可使膠質(zhì)瘤細(xì)胞發(fā)生G2/M期阻滯,其機(jī)制可能與CDK6、CDC25A表達(dá)的變化有關(guān);芹菜素可通過Bcl-2相關(guān)途徑誘導(dǎo)膠質(zhì)瘤SHG-44細(xì)胞凋亡。
[Abstract]:Objective to investigate the effects of apigenin at different concentrations on the proliferation inhibition and apoptosis induction of glioma SHG-44 cells. Methods SHG-44 glioma cells were treated with apigenin at different concentrations of 4080 渭 mol / L for 2448 ~ 72 h. The effects of apigenin at different concentrations on the proliferation inhibition rate of glioma cells were detected by MTT assay. The distribution of cell cycle in gliomas treated with apigenin for 24 h was observed by Pi single staining flow cytometry. The effect of apigenin at different concentrations on apoptosis of glioma cells was investigated by Hoechst 33258 Tunel staining and Annexin V-FITC / Pi double staining flow cytometry. The expression of cell cycle associated protein CDK6, CDC25A, and apoptosis-related protein BaxanBcl-2 were observed by Western blot. Results apigenin significantly inhibited the proliferation of SHG-44 cells and increased with the time and concentration of action. Cell cycle analysis showed that apigenin caused G2 / M phase arrest. Hoechst33258 immunofluorescence staining showed chromatin pyknosis in glioma cells treated with apigenin. Tunel immunofluorescence staining showed green fluorescence in apoptotic cells such as nuclear fragmentation and apoptotic bodies. Flow cytometry analysis showed that the apoptosis rate of SHG-44 cells increased with the increase of apigenin concentration. The results of 24 h treatment with different concentrations of apigenin showed that the drug could induce apoptosis of SHG-44 cells in a concentration-dependent manner. In apigenin treated group, the expression of CDK6 and CDC25A protein decreased gradually, the expression of Bax protein increased gradually, the expression of Bcl 2 protein decreased gradually and the ratio of Bax / Bax protein gradually increased. Conclusion apigenin can inhibit the proliferation of glioma SHG-44 cells in a time and dose-dependent manner. Apigenin can induce apoptosis of glioma SHG-44, apigenin can block G2 / M phase of glioma cells, and its mechanism may be related to the change of CDK6, CDC25A expression. Apigenin can induce apoptosis of glioma SHG-44 cells through Bcl-2 related pathway.
【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.41

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