本文選題：絲石竹皂甙　切入點：胃癌　出處：《青島大學》2016年碩士論文　論文類型：學位論文

【摘要】：目的:探究絲石竹提取物絲石竹皂甙在體外對人胃腺癌細胞株SGC-7901增殖、凋亡的影響及其機制。方法:取對數(shù)生長期的SGC-7901細胞作為研究對象,實驗組加入不同濃度(20、40、80ug/ml)的絲石竹皂甙培養(yǎng)液,以不加入絲石竹皂甙僅加入等體積培養(yǎng)基(含10%胎牛血清)的細胞作為對照組,分別培養(yǎng)24、48、72h。MTT法檢測細胞增殖活性;Annexin V/PI雙染法結合流式細胞儀分析細胞凋亡;流式細胞術DNA含量分析法分析細胞周期;實時熒光定量PCR檢測Wnt5a、Wnt8b基因的表達。結果:(1)①不同濃度的絲石竹皂甙作用相同時間,細胞存活率的差異具有統(tǒng)計學意義(p0.01)。24h細胞存活率:對照組為100.00%,實驗組(按濃度由低到高排列,下同)依次為(82.82±1.64)%、(50.58±1.04)%、(20.00±0.75)%,F=846.157;48h細胞存活率:對照組為100.00%,實驗組依次為(76.65±1.48)%、(40.28±1.40)%、(16.38±0.88)%,F=896.265;72h細胞存活率:對照組為100.00%,實驗組依次為(70.57±1.60)%、(27.47±1.21)%、(10.09±0.45)%,F=957.040。②相同濃度絲石竹皂甙作用不同時間(對照組除外),細胞存活率的差異具有統(tǒng)計學意義。當濃度為20ug/ml時,F=45.281,p0.01;濃度為40ug/ml時,F=212.674,p0.01;濃度為80ug/ml時,F=129.148,p0.01。(3)計算求得24小時IC50值為41.171ug/ml。(2)不同濃度的絲石竹皂甙作用24h后應用流式細胞術AV/PI雙染法進行細胞凋亡率分析:對照組為(15.11±0.95)%,實驗組依次為(20.33±0.78)%、(23.93±1.18)%、(27.67±1.44)%,F=83.215,p0.01。(3)不同濃度的絲石竹皂甙作用24h后應用流式細胞術DNA含量分析法進行細胞周期分析,結果以百分比表示,G0/G1期:對照組為(53.99±1.35)%,實驗組依次為(58.40±1.77)%、(62.40±0.85)%、(65.43±1.18)%,F=42.792,p0.01;S期:對照組為(31.64±1.30)%,實驗組依次為(27.07±1.38)%、(23.11±1.47)%、(19.81±0.70)%,F=53.396,p0.01。G2期:對照組為(14.71±0.06)%、實驗組依次為(14.91±0.18)%、(14.67±007)%、(14.71±0.17)%,F=2.209,p0.05。(4)不同濃度的絲石竹皂甙作用48h后應用實時熒光定量PCR進行基因表達分析,基因表達結果以相對比表示,Wnt5a/GAPDH1組:對照組為1.70±0.09,實驗組依次為1.22±0.10、0.74±0.13、0.58±0.07,F=109.516,p0.01;Wnt8b/GAPDH1組:對照組為1.18±0.08,實驗組依次為0.82±0.07、0.53±0.06、0.42±0.03,F=104.298,p0.01。結論:絲石竹皂甙可能通過下調Wnt基因的表達,在體外抑制胃癌細胞增殖、促進其凋亡,并使其細胞周期阻滯于G0/G1期,減少S期細胞比例。
[Abstract]:Objective: To explore the Gypsophila extract gypsoside in vitro on human gastric cancer cell SGC-7901 proliferation, apoptosis and its mechanism. Methods: SGC-7901 cells in logarithmic growth phase as the research object, different concentration in experimental group (20,40,80ug/ml) of gypsoside medium, to join gypsoside only adding volume (culture medium containing 10% fetal bovine serum) cells were cultured as the control group, 24,48,72h.MTT was used to detect the proliferation activity; analysis of apoptotic cells by flow cytometry with Annexin V/PI double staining method; flow cytometry DNA content analysis method to analyze the cell cycle; real time fluorescence quantitative PCR detection of Wnt5a, Wnt8b gene expression. Results: (1) the same time gypsoside effect of different concentration, the difference was statistically significant cell survival rate (P0.01):.24h cell survival was 100% in the control group, the experimental group (according to concentration by 浣庡埌楂樻帓鍒，

本文編號：1598718