發(fā)布時間：2018-01-13 14:11

  本文關(guān)鍵詞：COX-2、MMP-14和胰腺癌細(xì)胞增殖、侵襲、轉(zhuǎn)移的關(guān)系研究　出處：《石河子大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 胰腺癌 塞來昔布 基質(zhì)金屬蛋白酶14 增殖 侵襲 遷移

【摘要】：目的:通過測定不同濃度的環(huán)氧合酶-2抑制劑塞來昔布對胰腺癌細(xì)胞增殖、侵襲、遷移能力以及對環(huán)氧合酶-2(cyclooxygenase-2,COX-2)、基質(zhì)金屬蛋白酶14(matrix metalloproteinase14,MMP-14)的蛋白表達(dá)影響來探討COX-2抑制劑塞來昔布對胰腺癌的作用及相關(guān)作用機(jī)制。方法:1.體外培養(yǎng)人胰腺癌細(xì)胞系PANC-1細(xì)胞;2.分別用0(無)、20(低)、60(中)、100(高)μmol/L四個塞來昔布濃度對PANC-1細(xì)胞進(jìn)行不同時間(24、48和72h)處理,MTT比色法檢測細(xì)胞的增殖能力,Transwell侵襲實驗和劃痕實驗檢測細(xì)胞的侵襲能力和遷移能力,酶聯(lián)免疫吸附劑測定(EILISA)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.MTT結(jié)果顯示不同濃度的塞來昔布作用24、48和72h后PANC-1細(xì)胞的增殖能力下降,隨著塞來昔布濃度的增加,抑制作用逐漸增強(qiáng),且隨著塞來昔布作用時間的延長,抑制作用也逐漸加強(qiáng),抑制作用呈時間和濃度依賴性(P0.05);2.Transwell侵襲實驗結(jié)果顯示不同濃度的塞來昔布作用24h后PANC-1細(xì)胞的侵襲能力下降,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),呈濃度依賴性;3.劃痕實驗結(jié)果顯示不同濃度的塞來昔布作用24h后PANC-1細(xì)胞的遷移能力下降,相對遷移距離分別為11.400±0.959、9.511±1.110、5.856±0.778和2.844±0.416(F=59.516,P0.05),呈濃度依賴性;4.正常PANC-1細(xì)胞呈梭形或多角形,狀態(tài)良好,細(xì)胞形態(tài)學(xué)提示不同濃度的塞來昔布作用24h后PANC-1細(xì)胞逐漸變圓、皺縮并開始出現(xiàn)死細(xì)胞;5.ELISA結(jié)果提示不同濃度的塞來昔布作用24h后胰腺癌細(xì)胞中COX-2和MMP-14的蛋白表達(dá)水平相應(yīng)降低,均呈濃度依賴性,COX-2蛋白表達(dá)水平分別為0.876±0.023、0.682±0.027、0.505±0.022和0.384±0.019(F=262.537,P0.05);MMP-14蛋白表達(dá)水平分別為7.955±0.297、7.031±0.178、6.072±0.209、和5.047±0.216(F=89.247,P0.05);6.PANC-1細(xì)胞內(nèi)COX-2和MMP-14的蛋白表達(dá)具有顯著正相關(guān)性(r=0.873,P0.05)。結(jié)論:1.COX-2抑制劑塞來昔布以濃度梯度、時間梯度的形式減弱人胰腺癌細(xì)胞系PANC-1的增殖能力,以濃度梯度的形式減弱人胰腺癌細(xì)胞系PANC-1的侵襲及遷移能力;2.COX-2抑制劑塞來昔布抑制COX-2表達(dá)后,可能通過下調(diào)MMP-14表達(dá),進(jìn)而以濃度梯度形式減弱胰腺癌細(xì)胞的增殖、侵襲、遷移能力,起到抗胰腺癌作用。
[Abstract]:Objective: to determine the proliferation, invasion and migration of pancreatic cancer cells by celecoxib, a cyclooxygenase-2 inhibitor with different concentrations of cyclooxygenase-2 (Cyclooxygenase-2). Metalloproteinase 14 matrix metalloproteinase14. To investigate the effect of COX-2 inhibitor celecoxib on pancreatic cancer and its mechanism. Methods: 1. Human pancreatic cancer cell line PANC-1 was cultured in vitro. 2. Four celecoxib concentrations of 0 (0 ~ 0 ~ 20) (0 渭 mol/L) were used to treat the PANC-1 cells at different time intervals (n = 10). The concentration of celecoxib in PANC-1 cells was 100 渭 mol/L (high). 48 and 72 h) MTT colorimetric assay was used to detect the proliferation ability of the cells. Transwell invasion assay and scratch assay were used to detect the invasion ability and migration ability of the cells. The protein expression of COX-2 and MMP-14 was detected by Elisa. 3.The data were statistically analyzed. Results 1. The cell proliferation ability of PANC-1 cells was decreased after 24 h and 72 h treatment with different concentrations of celecoxib. With the increase of celecoxib concentration, the inhibitory effect was gradually enhanced, and with the prolongation of celecoxib action time, the inhibitory effect was gradually strengthened, and the inhibitory effect was time-and concentration-dependent (P0.05). 2. The results of transwell invasion experiment showed that the invasion ability of PANC-1 cells decreased after 24 h treatment with different concentrations of celecoxib. The number of cells passing through the matrix glue was 300.654 鹵12.558U 271.041 鹵10.569, respectively. 184.003 鹵10.865 and 92.667 鹵7.567 鹵7.567 鹵7.567FU 237.182P0.05, in a concentration-dependent manner. 3. The results of scratch test showed that the migration ability of PANC-1 cells was decreased after 24 h treatment with different concentrations of celecoxib, and the relative migration distance was 11.400 鹵0.959, respectively. 9.511 鹵1.110U 5.856 鹵0.778 and 2.844 鹵0.416FU 59.516P0.05U in a concentration-dependent manner. 4. The normal PANC-1 cells were fusiform or polygonal, and the morphology indicated that the PANC-1 cells gradually became round after 24 hours of celecoxib treatment with different concentrations of celecoxib. Shrink and begin to appear dead cells; 5. The results of Elisa showed that the protein expression of COX-2 and MMP-14 in pancreatic cancer cells decreased in a concentration dependent manner after 24 hours of treatment with different concentrations of celecoxib. The expression level of COX-2 protein was 0.876 鹵0.023 鹵0.682 鹵0.027, respectively. 0.505 鹵0.022 and 0.384 鹵0.019 respectively. The expression level of MMP-14 protein was 7.955 鹵0.297 鹵7.031 鹵0.178 鹵6.072 鹵0.209, respectively. And 5.047 鹵0.216F ~ (89) 247 (P _ (0.05)); 6. The expression of COX-2 and MMP-14 in PANC-1 cells was positively correlated with the expression of MMP-14. Conclusion 1. Cyclooxygenase-2 inhibitor celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in the form of concentration gradient and time gradient. The invasion and migration of human pancreatic cancer cell line PANC-1 were attenuated in the form of concentration gradient. 2. COX-2 inhibitor celecoxib inhibits the expression of COX-2, which may decrease the proliferation, invasion and migration of pancreatic cancer cells by down-regulating the expression of MMP-14, and then decreasing the proliferation, invasion and migration of pancreatic cancer cells in the form of concentration gradient. Play an anti-pancreatic cancer effect.
【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】：2015
【分類號】：R735.9

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本文編號：1419172