本文關(guān)鍵詞： 中藥艾迪 人喉癌Hep-2細胞 熒光定量PCR 環(huán)氧化酶-2　出處：《遼寧醫(yī)學(xué)院》2012年碩士論文　論文類型：學(xué)位論文

【摘要】：目的 研究中藥艾迪對人喉癌Hep-2細胞的生長抑制作用及誘導(dǎo)凋亡作用 方法 本研究設(shè)實驗組和對照組，實驗組為0.4、0.8、1.0、2.0ug/ml的中藥艾迪，對照組為未加藥物即0ug/ml。實驗組與對照組分別作用于人喉癌Hep-2細胞24h、48h及72h,四亞基噻唑藍（MTT）法測定中藥艾迪對人喉癌Hep-2細胞的生長抑制作用，通過Hoechst33258熒光染色后熒光顯微鏡下觀察人喉癌細胞Hep-2細胞的凋亡狀況，流式細胞儀（FCM）分析人喉癌Hep-2細胞的細胞周期及凋亡率的變化，熒光定量PCR法測定人喉癌Hep-2細胞中環(huán)氧化酶-2（COX-2）的表達量。 結(jié)果 ①MTT檢測示：實驗組對人喉癌Hep-2細胞的生長有抑制作用，與對照組比較，差異具有統(tǒng)計學(xué)意義（P＜0.05），并有明顯的時間和劑量依賴性。經(jīng)0.4、0.8、1.0、2.0ug/ml作用72h后，其細胞抑制率分別為23.08%、37.42%、43.38%、54.48%。②熒光檢測發(fā)現(xiàn)實驗組可見典型的凋亡細胞形態(tài)特征，可見凋亡小體，正常細胞明顯減少，，對照組未見明顯細胞凋亡形態(tài)變化，可見正常細胞核為藍色，染色質(zhì)均勻分布。③流式細胞儀檢測結(jié)果顯示實驗組作用72h后能增加G_0/G_1期的比例，減少S期和G_2/M的比例；實驗組作用后72h后的細胞凋亡率依次上升，由28.62±1.23%增加到54.90±2.40%，呈劑量依賴性，差異具有統(tǒng)計學(xué)意義（P＜0.01）。④熒光定量PCR法檢測，實驗組人喉癌Hep-2細胞所表達的環(huán)氧化酶-2（COX-2）水平呈劑量遞減關(guān)系。實驗組中2.0ug/ml作用人喉癌Hep-2細胞后所表達的環(huán)氧化酶-2（COX-2）水平最低，分別為對照組倍數(shù)的0.950±0.07、0.847±0.05、0.369±0.02。與對照組比較差異有統(tǒng)計學(xué)意義（P＜0.05）。 結(jié)論 中藥艾迪在體外能明顯抑制人喉癌細胞Hep-2生長增殖及誘導(dǎo)其凋亡，誘導(dǎo)細胞凋亡的機制可能與通過下調(diào)COX-2的表達有關(guān)。
[Abstract]:Purpose Study on the effect of Aidi on the growth inhibition and apoptosis of Hep-2 cells of Human Laryngeal carcinoma Method In this study, the experimental group and the control group, the experimental group is 0.4m 0.8U 1.0g / ml of Chinese medicine Aidi. The experimental group and the control group were treated with Hep-2 cells for 24 h and 72 h, respectively. The inhibitory effect of Aidi on the growth of human laryngeal cancer Hep-2 cells was determined by tetra-subunit thiazolium blue TTassay. The apoptosis of human laryngeal carcinoma cell line Hep-2 was observed under fluorescence microscope after Hoechst33258 fluorescence staining. Flow cytometry (FCM) was used to analyze the changes of cell cycle and apoptosis rate in human laryngeal carcinoma Hep-2 cells. The expression of cyclooxygenase-2 and cyclooxygenase-2 in human laryngeal carcinoma Hep-2 cells was determined by fluorescence quantitative PCR. Results 1MTT assay showed that the growth of human laryngeal carcinoma Hep-2 cells was inhibited in the experimental group, and the difference was statistically significant compared with the control group (P < 0.05). The cell inhibition rates were 23.08% and 37.42%, respectively, after 72 hours of treatment with 0.48 渭 g / ml of 0.48 渭 g / ml and 1.0 渭 g / ml of 0.48 渭 g / ml for 72 h, and in a time-and dose-dependent manner, the inhibition rate of the cells was 23.08%. Fluorescence detection of 43.38 and 54.48.2 found typical apoptotic cell morphological features, apoptotic bodies, normal cells decreased significantly in the experimental group, but no obvious changes in apoptotic morphology were found in the control group. The results showed that the normal nucleus was blue and chromatin distributed evenly. 3. The results of flow cytometry showed that after 72 hours of treatment, the proportion of G _ (0) / G _ (1) phase was increased, and the ratio of S phase to G _ 2 / M was decreased in the experimental group. The apoptosis rate of the experimental group increased from 28.62 鹵1.23% to 54.90 鹵2.40 in a dose-dependent manner. The difference was statistically significant (P < 0.01). 4 fluorescence quantitative PCR method was used to detect. Expression of cyclooxygenase-2mCOX-2 in human laryngeal carcinoma Hep-2 cells in experimental group. The level of cyclooxygenase-2 (COX-2) was the lowest in the experimental group (2.0ugml / ml) after the treatment of human laryngeal carcinoma Hep-2 cells. The ratio of the control group was 0.950 鹵0.07, 0.847 鹵0.05, 0.369 鹵0.02.Compared with the control group, the difference was statistically significant (P < 0.05). Conclusion Aidi can obviously inhibit the growth and proliferation of human laryngeal carcinoma cell line Hep-2 and induce its apoptosis in vitro. The mechanism of inducing apoptosis may be related to the down-regulation of COX-2 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】：2012
【分類號】：R739.65

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