發(fā)布時間：2018-04-10 02:42

  本文選題：食管癌ECa109細(xì)胞　切入點：塞來昔布　出處：《石河子大學(xué)》2017年碩士論文

【摘要】：目的:使用不同濃度COX-2抑制劑塞來昔布作用于食管癌ECa109細(xì)胞,測定細(xì)胞內(nèi)環(huán)氧合酶-2(cyclooxygenase-2,COX-2)和基質(zhì)金屬蛋白酶14(matrix metalloproteinase 14,MMP-14)的蛋白表達(dá),同時測定細(xì)胞增殖、侵襲、凋亡能力變化,從而進(jìn)一步研究COX-2、MMP-14相互作用在食管癌發(fā)生發(fā)展中的作用及機(jī)制。方法:1.體外培養(yǎng)人食管癌ECa109細(xì)胞;2.使用0(對照組)、20(低)、60(中)、100(高)μmol/L四個塞來昔布濃度對ECa109細(xì)胞進(jìn)行不同時間(24、48和72h)處理,MTT法檢測細(xì)胞的增殖能力,流式細(xì)胞儀測凋亡,Transwell侵襲實驗檢測細(xì)胞的侵襲能力,酶聯(lián)免疫法檢測COX-2和MMP-14的蛋白表達(dá)情況;3.數(shù)據(jù)進(jìn)行統(tǒng)計分析。結(jié)果:1.ELISA結(jié)果提示在塞來昔布濃度在0-100μmol/L之間,隨著藥物濃度遞增,細(xì)胞內(nèi)COX-2、MMP-14蛋白表達(dá)逐漸減少,均呈濃度依賴性,COX-2蛋白表達(dá)水平分別為1.581±0.116、1.226±0.089、0.846±0.076和0.521±0.082,四個組間差異有統(tǒng)計學(xué)意義(P0.05);MMP-14蛋白表達(dá)水平分別為11.517±0.881、7.905±0.724、5.788±0.750、和3.663±0.601,四個組間差異有統(tǒng)計學(xué)意義(P0.05);Pearson相關(guān)分析結(jié)果顯示兩者表達(dá)水平具有顯著正相關(guān)性(r=0.952,P0.05)。2.MTT檢測結(jié)果示不同濃度的藥物作用24、48和72h后,藥物對細(xì)胞增殖抑制率分別為:處理24 h后,較對照組(6.92±1.92)相比,20、60、100μmol/L組增殖率分別為25.52±2.39、40.60±4.06和49.42±2.67;處理48h后,較對照組(7.10±1.74)相比,20、60、100μmol/L組增殖率分別為43.70±5.20、55.77±4.18和66.44±4.03;處理72h后,較對照組(7.07±1.22)相比,20、60、100μmol/L組增殖率分別為60.34±4.64、72.09±3.93和84.31±3.21;隨著藥物濃度的提高,藥物對EC109細(xì)胞的增殖抑制率逐漸增強,且隨著藥物作用時間的增加,抑制作用也逐漸加強,抑制作用呈時間和濃度依賴性(P0.05);3 Transwell侵襲實驗結(jié)果顯示在不同濃度的藥物作用ECa109細(xì)胞24h后,細(xì)胞的侵襲能力下降,計數(shù)穿過基質(zhì)膠的細(xì)胞個數(shù)分別為300.654±12.558、271.041±10.569、184.003±10.865和92.667±7.567(F=237.182,P0.05),并呈濃度依賴性;4.流式細(xì)胞儀測定結(jié)果顯示在不同濃度的藥物作用ECa109細(xì)胞24h后,細(xì)胞凋亡率分別為1.700±0.557、13.400±1.735、18.767±1.301和28.300±71.988(F=161.843,P0.05)細(xì)胞的凋亡率逐漸增加,呈藥物濃度依賴性。結(jié)論:1.COX-2抑制劑塞來昔布可抑制食管癌ECa109細(xì)胞增殖,此作用具有濃度及時間依賴性;同時可促進(jìn)細(xì)胞的凋亡,抑制細(xì)胞的侵襲能力,此作用具有濃度依賴性。2.COX-2抑制劑塞來昔布可能通過抑制COX-2表達(dá),下調(diào)MMP-14表達(dá),從而抑制食管癌細(xì)胞的增殖、侵襲能力,促進(jìn)細(xì)胞凋亡,從而起到抗食管癌作用。
[Abstract]:Aim: to observe the expression of cyclooxygenase-2cyclooxygenase-2 (COX-2) and matrix metalloproteinase (14(matrix metalloproteinase 14) in esophageal carcinoma ECa109 cells treated with celecoxib at different concentrations, and to detect the changes of cell proliferation, invasion and apoptosis.To further study the role and mechanism of COX-2 MMP-14 interaction in the carcinogenesis and development of esophageal carcinoma.Method 1: 1.Human esophageal carcinoma ECa109 cells were cultured in vitro.The proliferation ability of ECa109 cells was detected by MTT assay with 0 (control group) (control group 0 (control group) with 4 celecoxib concentrations of 4 celecoxib at different time points, 24 渭 mol/L and 72 h), and the ability of cell invasion was detected by flow cytometry with apoptosis transwell invasion assay, and the cell proliferation was detected by flow cytometry (FCM), and the cell proliferation was detected by flow cytometry (FCM), and the proliferation of ECa109 cells was detected by flow cytometry.The protein expression of COX-2 and MMP-14 was detected by enzyme linked immunosorbent assay (Elisa).The data are analyzed statistically.Results 1. The results of Elisa showed that when the concentration of celecoxib ranged from 0 to 100 渭 mol/L, the expression of COX-2 MMP-14 protein gradually decreased with the increase of drug concentration.The results of MTT assay showed that the drug of different concentrations had a significant positive correlation after 24 h and 72 h, and the results of MTT assay showed that different concentrations of the drug could be used for 24 h, 48 h and 72 h, respectively.The inhibitory rates of the drug on cell proliferation were 25.52 鹵2.39 渭 mol/L and 49.42 鹵2.67, respectively, compared with the control group (6.92 鹵1.92) and the control group (43.70 鹵5.2055.77 鹵4.18 and 66.44 鹵4.03, respectively, 48 h after treatment), compared with the control group (7.10 鹵1.74), the proliferative rates were 43.70 鹵5.2055.77 鹵4.18 and 66.44 鹵4.03, respectively, after 48 hours of treatment, the cell proliferation inhibition rates were 43.70 鹵5.2055.77 鹵4.18 and 66.44 鹵4.03respectively.Compared with the control group (7.07 鹵1.22), the proliferative rates of the control group were 60.34 鹵4.64 渭 mol/L, 72.09 鹵3.93 and 84.31 鹵3.21, respectively. With the increase of the drug concentration, the inhibitory rate of the drug on EC109 cells was gradually increased, and the inhibitory effect was gradually strengthened with the increase of the time of the drug treatment.The results of flow cytometry showed that the apoptosis rates of ECa109 cells treated with different concentrations of drugs for 24 h were 1.700 鹵0.557 (13.400 鹵1.735N) 18.767 鹵1.301 and 28.300 鹵71.988FN 161.843 (P0.05), respectively, in a dose-dependent manner.Conclusion 1. Celecoxib, a COX-2 inhibitor, can inhibit the proliferation of esophageal carcinoma ECa109 cells in a concentration and time dependent manner, promote cell apoptosis and inhibit cell invasion.The effect is concentration-dependent. 2. Celecoxib, a COX-2 inhibitor, may inhibit the proliferation, invasion and apoptosis of esophageal cancer cells by inhibiting the expression of COX-2 and down-regulating the expression of MMP-14.
【學(xué)位授予單位】：石河子大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.1

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本文編號：1729335