本文選題：表沒(méi)食子兒茶素沒(méi)食子酸酯　切入點(diǎn)：鼻咽腫瘤　出處：《桂林醫(yī)學(xué)院》2010年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的初步探討綠茶提取物EGCG對(duì)鼻咽癌CNE2細(xì)胞增殖和凋亡的影響,并分析該過(guò)程中MnSOD、NF-κB、Caspase-3的表達(dá)變化,為可能尋找一個(gè)有效的增敏藥物與放療配合使用來(lái)提高鼻咽癌的治療效果。 方法1.體外培養(yǎng)鼻咽癌低分化株CNE2,用不同質(zhì)量濃度的(0、40、80、160mg/l)EGCG干預(yù)48h后,倒置顯微鏡下觀察CNE2細(xì)胞的形態(tài)學(xué)改變。2.不同質(zhì)量濃度的(0、40、80、160mg/l) EGCG作用CNE2細(xì)胞24、48、72h后,應(yīng)用MTT法間接檢測(cè)細(xì)胞增殖的變化,并計(jì)算抑制率。3.(0、40、80、160mg/l)EGCG作用CNE2細(xì)胞48h后,Hoechst33258染色觀察凋亡細(xì)胞,并計(jì)算凋亡率;流式細(xì)胞儀檢測(cè)EGCG對(duì)CNE2細(xì)胞周期的影響。4.RT-PCR檢測(cè)EGCG抗鼻咽癌細(xì)胞株CNE2過(guò)程中NF-κB、MnSOD、Caspase-3的表達(dá)變化。 結(jié)果1.EGCG處理CNE2細(xì)胞后,細(xì)胞分裂相變少,部分細(xì)胞變圓,隨著濃度的增加,部分細(xì)胞出現(xiàn)漂浮及壞死的現(xiàn)象。EGCG能抑制鼻咽癌細(xì)胞CNE2的增殖,隨著作用時(shí)間的延長(zhǎng)和濃度的增加抑制作用越明顯,表現(xiàn)為濃度與時(shí)間依賴性(P0.01)。2.EGCG能誘導(dǎo)細(xì)胞的凋亡,(0、40、80、160mg/l)EGCG處理CNE2細(xì)胞48h后,凋亡率分別為:(2.3±0.7)%、(13.7±1.1)%、(22.5±1.2)%、(36.2±2.1)%。與對(duì)照組比較,各實(shí)驗(yàn)組凋亡率明顯增加(P0.01),各實(shí)驗(yàn)組兩兩比較差異顯著(P0.01)。3.流式細(xì)胞術(shù)顯示:(0、40、80、160mg/l)EGCG作用CNE2后,停滯在Go/G1期細(xì)胞百分率分別為:(53.2±1.4)%、(64.8±1.2)%、(71.9±1.0)%、(74.5±1.6)%。與對(duì)照組比較,各實(shí)驗(yàn)組G1細(xì)胞百分率明顯增加(P0.01),各實(shí)驗(yàn)組兩兩比較差異顯著(P0.01),呈濃度依賴性。4. (0、40、80、160mg/l)EGCG處理CNE2細(xì)胞48h后,NF-κBmRNA的表達(dá)呈下降趨勢(shì);MnSOD和Caspase-3mRNA的表達(dá)呈升高趨勢(shì)。 結(jié)論1.EGCG能抑制鼻咽癌細(xì)胞CNE2的增殖,并表現(xiàn)為濃度和時(shí)間依賴性;EGCG能誘導(dǎo)鼻咽癌CNE2的凋亡,將細(xì)胞阻滯于G0/G1,呈現(xiàn)為濃度依賴性。2.EGCG可能通過(guò)下調(diào)NF-κBmRNA和上調(diào)MnSOD和Caspase-3的表達(dá)來(lái)發(fā)揮其抗鼻咽癌的生物學(xué)活性。
[Abstract]:Objective to investigate the effect of green tea extract (EGCG) on the proliferation and apoptosis of nasopharyngeal carcinoma (NPC) CNE2 cells, and to analyze the expression of MnSODNF- 魏 BnCaspase-3 in nasopharyngeal carcinoma (NPC) cells. Methods 1. The poorly differentiated nasopharyngeal carcinoma cell line CNE2 was cultured in vitro. After 48 hours of treatment with different concentrations of NCNE, the morphologic changes of CNE2 cells were observed under inverted microscope. The CNE2 cells were treated with different concentrations of Rhizoma 4080,160 mg / L EGCG for 72 h. The proliferation of CNE2 cells was detected indirectly by MTT assay, and the inhibition rate of CNE2 cells was calculated by Hoechst33258 staining for 48 h after treatment with 40,80mg / L EGCG, and the apoptotic rate was calculated. Flow cytometry was used to detect the effect of EGCG on the cell cycle of CNE2. 4. RT-PCR was used to detect the expression of Caspase-3 in EGCG anti-nasopharyngeal carcinoma cell line CNE2. Results 1. After treated with EGCG, the proliferation of nasopharyngeal carcinoma (NPC) cells was inhibited by CNE2. 2. After treated with EGCG, the cell division and transformation were less and some cells became round. With the increase of concentration, some cells appeared floating and necrotic. EGCG could inhibit the proliferation of nasopharyngeal carcinoma cell line CNE2. The inhibitory effect of EGCG was more obvious with the prolongation of time and the increase of concentration. 2. EGCG could induce apoptosis of CNE2 cells in a dose-dependent and time-dependent manner. The apoptotic rates of CNE2 cells treated with EGCG for 48 h were 2.3 鹵0.7 鹵1.21 鹵1.2 鹵2.10.Compared with the control group, the apoptosis rates of the cells were significantly higher than those of the control group (P < 0.05), but not in the control group (P < 0.05), and compared with that in the control group (P < 0.05), the apoptotic rate was significantly higher than that in the control group (P < 0.05), and the apoptosis rate was significantly higher than that in the control group (P < 0.05). The apoptotic rate of each experimental group was significantly increased, and the difference was significant between two groups. Flow cytometry showed that the percentage of cells stagnant in the Go/G1 phase after treatment with CNE2 was 53.2 鹵1.41 鹵1.21.9 鹵1.00.Compared with that of the control group, the percentage of cells in the Go/G1 phase was 74.5 鹵1.60.Compared with the control group, the percentage of cells in the Go/G1 phase was 53.2 鹵1.42 鹵1.21.The percentage of the cells in the control group was 74.5 鹵1.60.Compared with the control group, the percentage of cells in the Go/G1 phase was 53.2 鹵1.42 鹵1.21.9 鹵1.0mg / L, respectively. The percentage of G1 cells in each experimental group increased significantly (P 0.01), and the difference between the two groups was significant in a concentration dependent manner. The expression of NF- 魏 BmRNA in CNE2 cells increased in a dose-dependent manner after 48 hours of treatment with 100mg / L EGCG. Conclusion 1. EGCG can inhibit the proliferation of nasopharyngeal carcinoma (NPC) cell line CNE2 in a concentration and time dependent manner. EGCG can induce the apoptosis of CNE2 in nasopharyngeal carcinoma. 2. Blocking cells at G _ 0 / G _ 1 in a concentration-dependent manner. 2. EGCG may exert its biological activity against nasopharyngeal carcinoma by down-regulating NF- 魏 BmRNA and up-regulating the expression of MnSOD and Caspase-3.
【學(xué)位授予單位】：桂林醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R739.63

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