本文關(guān)鍵詞： 姜黃素 IAPS 信號(hào)通路 胃癌細(xì)胞 細(xì)胞凋亡　出處：《南華大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過(guò)觀察姜黃素對(duì)人胃癌細(xì)胞SGC-7901生長(zhǎng)的影響、細(xì)胞周期改變及NF-κB、Livin、Caspase-3表達(dá)的變化,初步探討姜黃素誘導(dǎo)胃癌細(xì)胞SGC-7901凋亡的可能機(jī)制。方法:(1)將對(duì)數(shù)生長(zhǎng)期的SGC-7901細(xì)胞,按每孔1000細(xì)胞加入到96孔細(xì)胞培養(yǎng)板中,接種容積為100μl;37℃、5%CO2條件下預(yù)貼壁12h后,分別加入濃度為10、20、40、60、80、100μmol/L姜黃素溶液、DMSO溶劑對(duì)照組及培養(yǎng)基空白對(duì)照組各100μl,每個(gè)濃度設(shè)置5個(gè)重復(fù)孔,分別培育12h、24h、36h、48h、72h后,用CCK-8法篩選最佳藥物濃度及作用時(shí)間;(2)設(shè)置空白對(duì)照組及20μmol/L,40μmol/L,60μmol/L姜黃素濃度組,將對(duì)數(shù)生長(zhǎng)期的SGC-7901細(xì)胞,按每孔5000細(xì)胞加入到6孔細(xì)胞培養(yǎng)板中,待細(xì)胞數(shù)量達(dá)到106/孔后,分別加入2m L1640培養(yǎng)基、20、40、60μmol/L姜黃素溶液,培育24h,用流式細(xì)胞儀測(cè)出各組細(xì)胞的細(xì)胞周期和凋亡率;(3)選取空白對(duì)照組及20μmol/L,40μmol/L,60μmol/L姜黃素濃度組,提取各組細(xì)胞的蛋白,Western Blot檢測(cè)各組SGC-7901細(xì)胞中NF-κB、Livin及Caspase-3等與胃癌細(xì)胞內(nèi)抗凋亡與促凋亡蛋白表達(dá)量的變化。(4)本實(shí)驗(yàn)所有實(shí)驗(yàn)結(jié)果采用SPSS18.0統(tǒng)計(jì)并進(jìn)行分析,對(duì)照組與實(shí)驗(yàn)組采用單因素配對(duì)樣本的t檢驗(yàn),方差分析兩組及兩組以上數(shù)據(jù)結(jié)果。結(jié)果:(1)CCK-8檢測(cè)結(jié)果:姜黃素藥物濃度增加以及作用時(shí)間的延長(zhǎng),SGC-7901細(xì)胞的增殖活力值逐漸下降,與對(duì)照組比較差異具有顯著性(P0.05)。Pearson關(guān)聯(lián)性分析結(jié)果顯示:與對(duì)照組比較SGC-7901細(xì)胞對(duì)不同濃度姜黃素具有劑量的依賴性(r=0.901,P=0.000.01)以及時(shí)間的依賴性(r=0.389,P=0.000.01)。(2)流式細(xì)胞儀檢測(cè)胃癌SGC-7901細(xì)胞的周期:姜黃素作用于SGC-7901細(xì)胞的藥物濃度的增加以及時(shí)間的延長(zhǎng),處于G1期的SGC-7901細(xì)胞數(shù)目明顯增加,且S期的細(xì)胞數(shù)目均下降。FCM檢測(cè)20μmol/L、40μmol/L、60μmol/L姜黃素溶液對(duì)SGC-7901凋亡率較對(duì)照組差異具有顯著性(P0.05)。(3)Western Blot免疫印跡分析蛋白表達(dá),加入姜黃素藥物處理SGC-7901細(xì)胞,隨著姜黃素濃度的增高,SGC-7901細(xì)胞內(nèi)NF-κB蛋白同Livin蛋白表達(dá)水平逐漸降低,Caspase-3蛋白表達(dá)水平逐漸升高。結(jié)論:1:姜黃素可明顯抑制體外培養(yǎng)的人胃癌SGC-7901細(xì)胞的增殖活力,促進(jìn)胃癌細(xì)胞凋亡。其抑制細(xì)胞增殖與促進(jìn)細(xì)胞凋亡具有明顯的濃度及時(shí)間依賴關(guān)系;2:姜黃素可能影響胃癌SGC-7901細(xì)胞周期,并阻滯在細(xì)胞周期G1期;3:姜黃素可能通過(guò)下調(diào)SGC-7901細(xì)胞內(nèi)NF-κB信號(hào)轉(zhuǎn)導(dǎo)蛋白及Livin蛋白表達(dá)水平,激活Caspase-3蛋白表達(dá),促進(jìn)SGC-7901細(xì)胞凋亡。
[Abstract]:Objective: the effect on the growth of human gastric cancer cell SGC-7901 by observing the changes of cell cycle and curcumin, NF- kappa B, Livin, Caspase-3 expression changes, discuss the possible mechanism of curcumin induced apoptosis of gastric cancer cell line SGC-7901. Methods: (1) the SGC-7901 cells in logarithmic growth phase at 1000 cells per well was added to the 96 hole cell culture plate, inoculation volume of 100 mu L; 37 DEG C, 5%CO2 under the condition of pre adherent after 12h were added at the concentration of 10,20,40,60,80100 mol/L curcumin solution, DMSO solvent control group and medium control group 100 L, each concentration set 5 repeated holes, respectively 24h, 36h, 12h training. 48h, 72h, the best screening time of drug concentration and action by CCK-8 method; (2) blank control group and 20 mol/L, 40 mol/L, 60 mol/L curcumin concentration group, the SGC-7901 cells in logarithmic growth phase at 5000 cells per well into the 6 Hole cell culture plate The number of cells, to achieve 106/ hole, we added the 2m L1640 medium, 20,40,60 mol/L curcumin solution, cultivation of 24h, using flow cytometry to measure cell apoptosis rate and cell cycle; (3) select the blank control group and 20 mol/L, 40 mol/L, 60 mol/L curcumin concentration group the cells were extracted, the protein of each group of SGC-7901 cells in Blot detection of Western NF- kappa B, and Caspase-3 and Livin in gastric cancer cells apoptosis and apoptosis protein expression. (4) in this experiment, all the experimental results using SPSS18.0 and statistical analysis, t test of control group and experimental group with single factor pair analysis of two groups of samples, and more than two sets of data variance. Results: (1) CCK-8 test results: the increase of curcumin concentration and the prolongation of time, the viability of SGC-7901 cells decreased, with significant differences compared with the control group (P0.0 5).Pearson correlation analysis showed that: compared with the control group of SGC-7901 cells in a dose dependent on different concentrations of curcumin (r=0.901, P=0.000.01) and time dependent (r=0.389, P=0.000.01). (2) flow cytometry SGC-7901 gastric cancer cell cycle: the increase of drug concentration in SGC-7901 cells by curcumin the role of the time and in the number of SGC-7901 cells in G1 phase increased significantly, and the number of cells in S phase decreased.FCM detection of 20 mol/L, 40 mol/L, 60 mol/L curcumin solution on the apoptosis rate of SGC-7901 was significant difference compared with the control group (P0.05). (3) the expression of Western Blot protein by Western blot analysis curcumin, drug treatment of SGC-7901 cells, with the increase of the concentration of curcumin in SGC-7901 cells, NF- kappa B protein and Livin protein expression decreased, Caspase-3 protein expression level increased gradually. Conclusion: ginger 1: Huang Suke significantly inhibited the in vitro cultured human gastric cancer SGC-7901 cell proliferation, promote apoptosis of gastric cancer cells. The inhibition of cell proliferation and promote cell apoptosis in a dose and time dependence of 2: obviously; curcumin may affect gastric cancer SGC-7901 cell cycle and arrest in the G1 phase of the cell cycle; 3: curcumin by inhibiting the expression of NF- B signal transduction protein and Livin protein level in SGC-7901 cells, activate the expression of Caspase-3 protein and promote the apoptosis of SGC-7901 cells.

【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張文陸;崔慧霞;王言;;復(fù)方苦參注射液對(duì)胃癌細(xì)胞SGC-7901凋亡的誘導(dǎo)作用及機(jī)制[J];廣東醫(yī)學(xué);2012年14期

2 高超;孔姝婧;趙晶;朱艷卿;裴東升;;雷帕霉素聯(lián)合紫杉醇對(duì)SGC-7901增殖影響的研究[J];中華腫瘤防治雜志;2014年02期

3 丁蘭;張世棟;劉國(guó)安;王瀚;楊東娟;王麗;;對(duì)映-貝殼杉烷二萜化合物Wangzaozin A對(duì)人胃腺癌細(xì)胞SGC-7901生長(zhǎng)的影響[J];西北師范大學(xué)學(xué)報(bào)(自然科學(xué)版);2006年03期

4 李迪諾;付曉光;;重組人血管內(nèi)皮抑制素對(duì)人胃癌細(xì)胞株SGC-7901表達(dá)VEGF的抑制作用[J];藥物生物技術(shù);2010年04期

5 許慶瑞;張樹(shù)明;張俊威;徐琛;王偉明;;復(fù)方青龍衣膠囊對(duì)胃癌細(xì)胞SGC-7901基因芯片表達(dá)的影響[J];中國(guó)實(shí)驗(yàn)方劑學(xué)雜志;2011年08期

6 姜成鋼;李嘉斌;徐惠綿;于淼;吳濤;劉芙蓉;;西咪替丁對(duì)胃癌細(xì)胞SGC-7901增殖及凋亡的影響[J];世界華人消化雜志;2007年02期

7 楊小珊,曾令福;葉酸對(duì)體外人胃腺癌細(xì)胞SGC-7901生長(zhǎng)的作用[J];中國(guó)公共衛(wèi)生;2003年12期

8 沈克平;王海永;胡兵;劉威;;胃腸安對(duì)SGC-7901胃癌上皮間質(zhì)轉(zhuǎn)化相關(guān)基因表達(dá)影響[J];時(shí)珍國(guó)醫(yī)國(guó)藥;2012年04期

9 王崴;于蕾;崔榮田;莫科;鄒翔;;野西瓜揮發(fā)油誘導(dǎo)SGC-7901凋亡的初步研究[J];哈爾濱商業(yè)大學(xué)學(xué)報(bào)(自然科學(xué)版);2008年04期

10 ;[J];;年期

相關(guān)會(huì)議論文 前1條

1 黃正華;鄒移海;王茵萍;黃冰;蘇喬;張薇;謝玲玲;陳嘉;;川芎嗪對(duì)SGC-7901胃腺癌細(xì)胞VEGF和bFGF分泌的影響[A];中國(guó)實(shí)驗(yàn)動(dòng)物學(xué)會(huì)第七屆學(xué)術(shù)年會(huì)論文集[C];2006年

相關(guān)碩士學(xué)位論文 前4條

1 黃果;姜黃素誘導(dǎo)胃癌細(xì)胞SGC-7901凋亡機(jī)制的研究[D];南華大學(xué);2015年

2 張瀛;高純度雄黃對(duì)胃腺癌細(xì)胞株SGC-7901凋亡的影響[D];華中科技大學(xué);2013年

3 丁蕓霞;附子生物堿對(duì)胃癌細(xì)胞株SGC-7901的抑制作用及其與化療藥物協(xié)同作用的實(shí)驗(yàn)研究[D];南京中醫(yī)藥大學(xué);2014年

4 陳建新;無(wú)血清懸浮培養(yǎng)法富集人胃癌SGC-7901干細(xì)胞及干性鑒定的研究[D];福建醫(yī)科大學(xué);2014年

，

本文編號(hào)：1534072