本文關(guān)鍵詞：哈薩克藥阿爾泰瑞香不同提取物對(duì)人食管癌Eca-109細(xì)胞的體外抑制作用　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 哈薩克藥 阿爾泰瑞香 人食管癌Eca-109細(xì)胞 細(xì)胞周期 細(xì)胞凋亡 PPARγ基因

【摘要】：目的:采用現(xiàn)代細(xì)胞分子生物學(xué)實(shí)驗(yàn)技術(shù),初探阿爾泰瑞香提取物抗食管癌Eca-109細(xì)胞作用機(jī)制,為哈薩克藥阿爾泰瑞香的后期研究提供依據(jù)。方法:將阿爾泰瑞香三種提取物不同濃度,依次作用于人食管癌Eca-109細(xì)胞,采用四甲基偶氮唑鹽法(MTT法)測(cè)定Eca-109細(xì)胞的增殖抑制率;通過(guò)計(jì)算細(xì)胞增殖抑制率,分析阿爾泰瑞香提取物對(duì)人食管癌細(xì)胞系Eca-109細(xì)胞增殖活性的影響。應(yīng)用倒置熒光顯微鏡技術(shù),觀察細(xì)胞生長(zhǎng)狀況及形態(tài)學(xué)變化。運(yùn)用流式細(xì)胞術(shù),檢測(cè)細(xì)胞周期及細(xì)胞凋亡率,探討阿爾泰瑞香對(duì)人食管癌Eca-109細(xì)胞的增殖抑制的作用及機(jī)制。采用蛋白免疫印跡技術(shù)及實(shí)時(shí)熒光定量PCR技術(shù)對(duì)細(xì)胞內(nèi)的PPARγ基因的表達(dá)水平進(jìn)行分析,探討阿爾泰瑞香不同提取物抗人食管癌Eca-109細(xì)胞的作用機(jī)制及與基因表達(dá)調(diào)控的關(guān)系。結(jié)果:阿爾泰瑞香三種提取物對(duì)人食管癌Eca-109細(xì)胞具有明顯的抑制細(xì)胞生長(zhǎng)的作用,與對(duì)照組比較,差異有統(tǒng)計(jì)學(xué)意義(P㩳0.05)。人食管癌Eca-109細(xì)胞經(jīng)阿爾泰瑞香三種提取物處理后細(xì)胞形態(tài)發(fā)生變化。用阿爾泰瑞香三種提取物不同濃度處理人食管癌Eca-109細(xì)胞后處于S期的細(xì)胞數(shù)顯著升高,跟空白對(duì)照組比較,差異具有統(tǒng)計(jì)學(xué)意義(P㩳0.05);G0/G1期的細(xì)胞數(shù)明顯下降,與空白對(duì)照組比較,差異亦具有統(tǒng)計(jì)學(xué)意義(P㩳0.05)。人食管癌Eca-109細(xì)胞經(jīng)阿爾泰瑞香三種提取物不同濃度處理后,檢測(cè)細(xì)胞凋亡率,結(jié)果與空白對(duì)照組比較藥物干預(yù)后癌細(xì)胞凋亡率明顯增大,差異有統(tǒng)計(jì)學(xué)意義(P㩳0.05)。人食管癌Eca-109細(xì)胞經(jīng)阿爾泰瑞香提取物不同濃度處理不同時(shí)間(24 h,48 h,72 h)后采用熒光定量PCR法檢測(cè)細(xì)胞內(nèi)PPARγmRNA表達(dá)量,結(jié)果經(jīng)三種提取物處理癌細(xì)胞后細(xì)胞內(nèi)PPARγmRNA表達(dá)量均顯著增加(P㩳0.05)。阿爾泰瑞香乙酸乙酯提取物干預(yù)24 h,48 h,72 h后細(xì)胞內(nèi)PPARγmRNA表達(dá)量增高(P㩳0.05)。阿爾泰瑞香正己烷、甲醇提取物干預(yù)48 h、72 h細(xì)胞內(nèi)PPARγmRNA表達(dá)量增高(P㩳0.05),而干預(yù)24 h后PPARγmRNA表達(dá)量未見明顯變化(P0.05)。Western Blot結(jié)果顯示經(jīng)阿爾泰瑞香提取物干預(yù)后細(xì)胞內(nèi)PPARγ蛋白表達(dá)增高(P㩳0.05),干預(yù)48 h后的上調(diào)作用較強(qiáng)。結(jié)論:阿爾泰瑞香提取物抑制人食管癌Eca-109細(xì)胞增殖,阻滯細(xì)胞周期,誘導(dǎo)細(xì)胞凋亡,上調(diào)細(xì)胞內(nèi)PPARγ基因表達(dá)。
[Abstract]:Objective: to explore the mechanism of anti-esophageal cancer Eca-109 cells by using modern cell molecular biological techniques. Methods: three kinds of extracts of Artemisia artemiana were used in human esophageal carcinoma Eca-109 cells in different concentrations. The proliferation inhibition rate of Eca-109 cells was measured by MTT assay. By calculating the inhibition rate of cell proliferation, the effect of the extract of Artemisia tenuifolia on the proliferation of human esophageal carcinoma cell line Eca-109 was analyzed. The inverted fluorescence microscopy was used. Cell cycle and apoptosis rate were detected by flow cytometry. To investigate the inhibitory effect and mechanism of Artemisia on the proliferation of human esophageal carcinoma Eca-109 cells. Expression of PPAR 緯 gene in human esophageal carcinoma Eca-109 cells by Western blot and real-time fluorescence quantitative PCR. Level analysis. To investigate the mechanism of different extracts of Daphne artemiana on human esophageal carcinoma Eca-109 cells and its relationship with gene expression regulation. The three extracts of Artemisia artemiana have obvious inhibitory effect on the growth of human esophageal carcinoma Eca-109 cells. Compared with the control group, the difference was statistically significant. 0.05). The morphological changes of human esophageal carcinoma Eca-109 cells treated with three extracts of Artemisia artemianum were observed in different concentrations of human esophageal carcinoma Eca-109 cells. The number of cells in S phase was significantly increased. Compared with the blank control group, the difference was statistically significant. The number of cells in the G _ 0 / G _ 1 phase of 0.05G _ 0 / G _ 1 decreased significantly, and the difference was statistically significant compared with the blank control group. The apoptosis rate of human esophageal carcinoma Eca-109 cells was measured after treated with different concentrations of three extracts of Artemisia tenuifolia. Results compared with the blank control group, the apoptosis rate of cancer cells increased significantly after drug intervention, and the difference was statistically significant. Human esophageal carcinoma Eca-109 cells were treated with different concentrations of Artemisia tenuifolia extract for 24 h or 48 h. After 72 h, the expression of PPAR 緯 mRNA was detected by fluorescence quantitative PCR assay. Results after treated with three kinds of extracts, the expression of PPAR 緯 mRNA in cancer cells increased significantly. The expression of PPAR 緯 mRNA in the cells was increased after treatment with ethyl acetate extract for 24 h and 48 h for 72 h. The expression of PPAR 緯 mRNA in the cells was increased after 48 h of treatment with methanol extract. 0.05). However, the expression of PPAR 緯 mRNA did not change significantly after 24 h intervention (P 0.05). The results of Western Blot showed that the expression of PPAR 緯 protein in the cells was increased after the intervention of the extract of Artemisia tenuifolia. Conclusion: the extract of Daphne altae inhibits the proliferation of human esophageal carcinoma Eca-109 cells, blocks cell cycle and induces apoptosis. The expression of PPAR 緯 gene was up-regulated.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R29

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 曹小琴;孫喜斌;;食管癌發(fā)病水平及變化趨勢(shì)[J];中國(guó)腫瘤臨床;2016年21期

2 戈偉;閆亞飛;;食管癌的分子靶向治療進(jìn)展[J];醫(yī)學(xué)與哲學(xué)(B);2016年10期

3 木拉提·克扎衣別克;夏木西努爾·玉山;蘇建春;;哈薩克藥阿爾泰瑞香及同屬植物的傳統(tǒng)應(yīng)用及抗癌活性研究進(jìn)展[J];河北醫(yī)藥;2016年19期

4 張思維;鄭榮壽;左婷婷;曾紅梅;陳萬(wàn)青;赫捷;;中國(guó)食管癌死亡狀況和生存分析[J];中華腫瘤雜志;2016年09期

5 凱德麗艷·阿布都外力;張慧霞;李旭峰;陳丹;陳艷;;葉酸在MNNG致哈薩克族食管上皮細(xì)胞增殖、周期及凋亡中的作用[J];新疆醫(yī)科大學(xué)學(xué)報(bào);2016年03期

6 李維;李晨希;于紅剛;;食管癌侵襲與轉(zhuǎn)移過(guò)程中相關(guān)蛋白研究進(jìn)展[J];疑難病雜志;2015年11期

7 趙艷平;楊春旭;;多烯紫杉醇對(duì)人食管癌EC-109細(xì)胞增殖及凋亡的影響[J];中國(guó)老年學(xué)雜志;2015年19期

8 俞宗佑;楊孝樸;;DAPI染色法在流式細(xì)胞術(shù)中檢測(cè)BHK-21細(xì)胞周期的探討[J];安徽農(nóng)業(yè)科學(xué);2015年23期

9 王穎婉;李葉麗;王俊逸;華亮;楊丹莉;;PPARα、PPARγ的上調(diào)參與淫羊藿苷對(duì)自發(fā)性高血壓大鼠心室重構(gòu)的調(diào)節(jié)[J];中國(guó)藥理學(xué)通報(bào);2015年08期

10 善杜哈喜·巴哈提江;木拉提·克扎衣別克;地達(dá)爾·巴合提堅(jiān);帕娜爾·沙吾提巴依;熱依扎·哈斯木汗;阿依努爾·居馬拜;;阿爾泰瑞香醇化學(xué)成分提物及其對(duì)癌細(xì)胞凋亡的誘導(dǎo)作用[J];天津醫(yī)藥;2015年02期

相關(guān)會(huì)議論文 前1條

1 李思蒙;O垂鸚，

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