【摘要】：目的 研究紫草素對人喉癌Hep-2細胞增殖和凋亡的作用。 方法 以不同濃度的紫草素作用于人喉癌Hep-2細胞, MTT法檢測紫草素對人喉癌Hep-2細胞增殖的抑制作用,熒光顯微鏡觀察人喉癌Hep-2細胞凋亡狀況,流式細胞儀分析細胞周期分布及凋亡率變化,熒光定量PCR法測量人喉癌Hep-2細胞增殖誘導(dǎo)配體(a proliferation-inducing ligand,APRIL)的表達量。 結(jié)果 紫草素對人喉癌Hep-2細胞的增殖有明顯抑制作用,并呈明顯的量效關(guān)系和時間依賴性。紫草素促進人喉癌Hep-2細胞凋亡。Hoechst 33258熒光檢測發(fā)現(xiàn)對照組可見正常細胞核為藍色,染色質(zhì)均勻分布;實驗組可見典型的凋亡細胞形態(tài)特征,可見凋亡小體;流式細胞儀檢測結(jié)果顯示:不同濃度紫草素作用48h后的細胞凋亡率依次上升,呈劑量依賴性,細胞周期呈G2/M期阻滯。紫草素作用人喉癌Hep-2細胞后其APRIL的mRNA表達量隨時間的延長和濃度的增加而升高,于作用后12h-48h各不同濃度組與空白組比較差異有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論 紫草素可以抑制喉癌Hep-2細胞的增殖,誘導(dǎo)人喉癌Hep-2細胞凋亡。紫草素作用人喉癌Hep-2細胞后未凋亡的細胞因APRIL的表達升高而有增殖能力增強的潛在趨勢。
[Abstract]:Objective to study the effect of Shikonin on proliferation and apoptosis of human laryngeal carcinoma Hep-2 cells. Methods Human laryngeal carcinoma Hep-2 cells were treated with different concentrations of porphyrin. The inhibitory effect of shikonin on the proliferation of human laryngeal carcinoma Hep-2 cells was detected by MTT assay. The apoptosis of Hep-2 cells was observed by fluorescence microscope. The changes of cell cycle distribution and apoptosis rate were analyzed by flow cytometry. The expression of a proliferation-inducing ligand,APRIL was measured by fluorescence quantitative PCR in human laryngeal carcinoma Hep-2 cells. Results Shikonin inhibited the proliferation of human laryngeal carcinoma Hep-2 cells in a dose-dependent and time-dependent manner. The results of fluorescence detection showed that the normal nucleus was blue and chromatin was uniformly distributed in the control group, and typical apoptotic cells and apoptotic bodies were found in the experimental group. The results of flow cytometry showed that the apoptosis rate increased in a dose-dependent manner after 48 h treatment with different concentrations of porphyrin, and the cell cycle was blocked in G _ 2 / M phase. The expression of APRIL mRNA in human laryngeal carcinoma Hep-2 cells increased with the prolongation of time and the increase of the concentration after the treatment of Shikonin, and there was significant difference between the different concentration groups of 12h-48h and the blank group (P0.05). Conclusion Shikonin can inhibit the proliferation of Hep-2 cells and induce apoptosis of human laryngeal carcinoma Hep-2 cells. The unapoptotic Hep-2 cells of human laryngeal carcinoma have the potential tendency of proliferation enhancement due to the increase of APRIL expression.
【學(xué)位授予單位】：遼寧醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R739.65

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