本文選題：表沒食子兒茶素沒食子酸酯 + caspase-8�。� 參考：《皖南醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：探討綠茶提取物表沒食子兒茶素沒食子酸酯（epigallocathechin-3-gallate,EGCG）誘導(dǎo)口腔鱗癌Tca8113細胞凋亡及其可能機制研究。以期為EGCG在口腔鱗癌預(yù)防和治療中提出理論依據(jù)。 方法：不同劑量的EGCG（0、25、50、75、100、125mg/L）分別作用Tca8113細胞24h、48h、72h,MTT法觀察該藥物對Tca8113細胞生長的影響，并計算細胞增殖抑制率。倒置顯微鏡下觀察EGCG（0、50、75、100mg/L）處理48h后Tca8113細胞的形態(tài)學(xué)變化。AO/EB熒光染色法檢測不同劑量的EGCG（0、25、50、75、100mg/L）分別作用Tca8113細胞48h后熒光顯微鏡下細胞的形態(tài)學(xué)變化。Hochest33258染色法檢測不同劑量EGCG（0、25、50、75、100、125mg/L）對Tca8113細胞凋亡的影響。流式細胞儀檢測EGCG（0、50、75mg/L）干預(yù)48h后Tca8113細胞周期的改變。透射電鏡下觀察加藥組與陰性對照組之間的區(qū)別，其中加藥組是否出現(xiàn)凋亡現(xiàn)象。免疫細胞化學(xué)SABC法檢測不同濃度的EGCG（0、25、50、75、100、125mg/L）干預(yù)48h后，caspase-3、caspase-8蛋白在Tca8113細胞中的表達差異。 結(jié)果：MTT法檢測結(jié)果顯示，不同劑量的EGCG（25、50、75、100、125mg/L）均能抑制口腔鱗癌Tca8113細胞的增殖，其中（50、75、100、125mg/L）與對照組比較差異有統(tǒng)計學(xué)意義（p0.05），細胞增值抑制率隨著EGCG濃度的增加和作用時間的延長而升高，呈現(xiàn)出明顯的時間-劑量效應(yīng)關(guān)系。倒置顯微鏡下可見對照組的Tca8113細胞呈梭形或多邊形貼壁生長，數(shù)量密集，輪廓清楚，細胞間連接緊密。加藥組的Tca8113細胞隨著EGCG濃度的增加，貼壁的細胞數(shù)量明顯減少，折光性差，部分細胞變圓、皺縮并漂浮于培養(yǎng)基中。AO/EB熒光染色法顯示，，不同濃度的EGCG干預(yù)Tca8113細胞中出現(xiàn)了典型的凋亡形態(tài)學(xué)變化，與對照組比較，隨著EGCG濃度的增加，凋亡細胞數(shù)不斷增加。Hochest33258染色顯示，不同濃度的EGCG干預(yù)Tca8113細胞中出現(xiàn)了典型的凋亡形態(tài)學(xué)改變?nèi)绾藵饪s、核碎裂等。與對照組比較，隨著EGCG濃度的增加，凋亡細胞數(shù)不斷增加（n=5,p0.01）。分別采用不同濃度EGCG（0、50、75mg/L）處理Tca8113細胞48h后通過流式細胞儀進行細胞周期分析，結(jié)果顯示EGCG（50、75mg/L）處理組G1期細胞百分數(shù)分別為41.997±0.17和65.24±0.632，與對照組（未加藥組）比較具有顯著性差異（p0.01），表明EGCG誘導(dǎo)Tca8113細胞G1期阻滯。透射電鏡下發(fā)現(xiàn)與對照組相比，加藥組（75mg/L）出現(xiàn)典型的凋亡改變。免疫細胞化學(xué)檢測結(jié)果顯示，EGCG能上調(diào)Tca8113細胞中caspase-3、caspase-8蛋白的表達，且呈濃度依賴關(guān)系。不同濃度的EGCG（0,、25、50、75、100mg/L）干預(yù)Tca8113細胞48h，caspase-8蛋白的平均光密度（AOD）值分別為：0.131±0.04、0.177±0.019、0.223±0.06、0.376±0.074、0.571±0.085，各加藥組與空白對照組相比差異有統(tǒng)計學(xué)意義（p0.01），caspase-3蛋白的平均光密度（AOD）值分別為：0.096±0.072、0.118±0.049、0.197±0.081、0.343±0.033、0.653±0.065，各加藥組與空白對照組相比差異有統(tǒng)計學(xué)意義（p0.05）。 結(jié)論：EGCG能有效地抑制Tca8113細胞的增殖，誘導(dǎo)其凋亡，其機制可能與上調(diào)caspase-8、caspase-3的表達有關(guān)。
[Abstract]:Aim: to investigate the apoptosis of oral squamous cell carcinoma (Tca8113) cells induced by epigallocathechin-3-gallate EGCG, an epigallocathin-3-gallateEGCG extract from green tea extract. To provide a theoretical basis for EGCG in the prevention and treatment of oral squamous cell carcinoma. Methods: Tca8113 cells were treated with different doses of EGCG 2550U 75100125 mg / L for 24 h, 48 h and 72 h, respectively. The effect of the drug on the growth of Tca8113 cells was observed and the cell proliferation inhibition rate was calculated. Morphologic changes of Tca8113 cells after 48 h treatment with EGCG 050,75100mg / L) .AOP / EB fluorescence staining method was used to detect the morphological changes of Tca8113 cells treated with different dosages of EGCGG 02550mg / L respectively for 48 h. Hochest33258 staining method was used to detect the morphologic changes of Tca8113 cells with different doses of EGCG02550Mg / L (75100125mg / L). The effect on apoptosis of Tca8113 cells. Flow cytometry was used to detect the changes of Tca8113 cell cycle after treatment with EGCG 50 mg / L for 48 h. Transmission electron microscope was used to observe the difference between the drug adding group and the negative control group. The expression of caspase-3 and caspase-8 protein in Tca8113 cells was detected by immunocytochemistry SABC method. Results the cell proliferation of oral squamous cell carcinoma (Tca8113) cells was inhibited by different doses of EGCG2550575100125mg / L, and there was a significant difference between the control group and the control group (P 0.05). The cell proliferation inhibition rate increased with the increase of EGCG concentration and the prolongation of the time of action. There is an obvious time-dose-effect relationship. Under the inverted microscope, the Tca8113 cells in the control group were spindle-shaped or polygonal adherent, dense in number, clear in outline and closely connected between cells. With the increase of EGCG concentration, the number of adherent cells decreased significantly, some cells became round, shrinked and floated in the medium with fluorescence staining. Typical morphological changes of apoptosis appeared in Tca8113 cells treated with different concentrations of EGCG. Compared with the control group, the number of apoptotic cells increased with the increase of EGCG concentration. Hochest33258 staining showed that the number of apoptotic cells increased with the increase of EGCG concentration. Typical apoptotic morphological changes such as nuclear concentration and nuclear fragmentation appeared in Tca8113 cells treated with different concentrations of EGCG. Compared with the control group, the number of apoptotic cells increased with the increase of EGCG concentration. Tca8113 cells were treated with different concentrations of EGCG 50 mg / L for 48 h, and the cell cycle was analyzed by flow cytometry. The results showed that the percentage of G 1 phase cells in EGCG 50 mg / L group was 41.997 鹵0.17 and 65.24 鹵0.632, respectively, which was significantly different from that in control group (p0.01), indicating that EGCG induced Tca8113 cell G1 phase arrest. Compared with the control group, 75 mg / L of apoptosis was observed under TEM. The results of immunocytochemistry showed that EGCG could up-regulate the expression of caspase-3 and caspase-8 in Tca8113 cells in a concentration-dependent manner. The average optical density of caspase-8 protein was 0.131 鹵0.04 鹵0.177 鹵0.019 鹵0.223 鹵0.076 鹵0.074 鹵0.571 鹵0.085, respectively. The average optical density of caspase-8 protein was 0.096 鹵0.072 鹵0.118 鹵0.049 鹵0.197 鹵0.033 鹵0.653 鹵0.0655.Compared with the blank control group, the average optical density of caspase-3 protein was 0.096 鹵0.072 鹵0.118 鹵0.049 鹵0.197 鹵0.081 鹵0.343 鹵0.033 鹵0.653 鹵0.0655.Compared with the blank control group, the average optical density of caspase-3 protein in each drug group was significantly higher than that in the blank control group. The average optical density of caspase-8 protein was 0.096 鹵0.072 鹵0.118 鹵0.049 鹵0.197 鹵0.081 鹵0.343 鹵0.033 鹵0.653 鹵0.065, respectively. The difference between the two groups was statistically significant (P 0.05). ConclusionEGCG can effectively inhibit the proliferation and induce apoptosis of Tca8113 cells, which may be related to the up-regulation of caspase-8 and caspase-3 expression.
【學(xué)位授予單位】：皖南醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R739.85

【相似文獻】

相關(guān)期刊論文 前10條

1 丁平平;伍睿;何雯;;兒茶素單體EGCG研究的文獻統(tǒng)計分析[J];科技信息(學(xué)術(shù)研究);2008年27期

2 陳曉燕;鄧啟盼;羅非君;曹亞;;EGCG在腫瘤預(yù)防及治療中作用的分子機制[J];生命的化學(xué);2011年02期

3 寧喜;倪菊芳;易世江;;EGCG聯(lián)合放療治療鼻咽癌裸鼠移植瘤模型[J];中國耳鼻咽喉頭頸外科;2011年04期

4 歐冰凝;周煥第;梁鋼;孫悅文;;EGCG衍生物在高溫和低溫強光照射下的穩(wěn)定性研究[J];廣西醫(yī)科大學(xué)學(xué)報;2012年05期

5 楊曉;李彩蓉;朱忠華;蘇華;盧遠航;;EGCG對實驗性糖尿病大鼠腎臟的保護作用[J];中國藥理學(xué)通報;2006年01期

6 張素梅;楊松;黃榮茂;;茶提取物EGCG抗腫瘤活性及機理研究進展[J];貴州農(nóng)業(yè)科學(xué);2006年S1期

7 蔡飛;李彩蓉;吳基良;陳金和;;EGCG對局灶性腦缺血損傷的作用研究[J];食品科學(xué);2006年09期

8 朱斌;陳曉光;;二次柱層析制備高純度表沒食子兒茶素沒食子酸酯(EGCG)的工藝研究[J];食品與機械;2009年04期

9 朱美霖;陸潔;韓梅;;表沒食子兒茶素沒食子酸酯(EGCG)與人血清白蛋白相互作用的研究[J];北京師范大學(xué)學(xué)報(自然科學(xué)版);2011年03期

10 蔣潔琳;溫旭燁;胡雅瓊;羅理勇;肖海軍;曾亮;;表沒食子兒茶素沒食子酸酯(EGCG)對癌癥細胞信號傳導(dǎo)鏈的影響[J];食品科學(xué);2012年09期

相關(guān)會議論文 前10條

1 鄭紅發(fā);黃亞輝;黃懷生;;高EGCG茶的研究與開發(fā)[A];湖南省茶葉優(yōu)勢區(qū)域建設(shè)會議暨湖南省茶葉學(xué)會2006年學(xué)術(shù)年會論文集[C];2006年

2 杜琪珍;沈星榮;蔣迎;;茶葉中EGCG的制備化分離新技術(shù)[A];海峽兩岸茶葉科技學(xué)術(shù)研討會論文集[C];2000年

3 陳偉強;李樹田;侯s

本文編號：1990288