本文選題：青藤堿　切入點：人胃癌細胞SGC-7901　出處：《河北醫(yī)科大學》2015年碩士論文

【摘要】：目的:研究青藤堿(SIN)對人胃癌細胞SGC-7901增殖、侵襲以及轉(zhuǎn)移相關基因和蛋白表達的影響。方法:1細胞存活率檢測:用不同濃度SIN處理人胃癌SGC-7901細胞后,用WST-1法檢測不同處理時間的細胞存活率。2 MMP表達:用不同濃度SIN處理人胃癌SGC-7901細胞,ELISA檢測MMP-2和MMP-9在細胞培養(yǎng)基中的含量變化。3 RT-PCR檢測不同濃度SIN對人胃癌SGC-7901細胞相關EMT基因mRNA表達水平的影響。4 Western blotting檢測不同濃度SIN對人胃癌SGC-7901細胞相EMT蛋白表達水平的影響。5 Western blotting檢測不同濃度SIN對人胃癌SGC-7901細胞核內(nèi)NFκB p65蛋白表達水平的影響。結果:1采用不同濃度的SIN處理人胃癌SGC-7901細胞,檢測結果表明:處理24 h后,1 mmol/L、2 mmol/L、2.5 mmol/L和3mmol/L處理組的細胞存活率顯著低于對照組(P0.05)。處理48 h后,0.5 mmol/L、1 mmol/L、2 mmol/L、2.5 mmol/L以及3 mmol/L處理組的細胞存活率顯著低于對照組(P0.05),且對青藤堿具有時間和劑量依賴性。2 SIN處理后,可降低人胃癌SGC-7901細胞培養(yǎng)基MMPs的含量。各SIN處理組培養(yǎng)基中MMP-2含量顯著低于對照組(P0.05),1 mmol/L和2 mmol/L處理組的MMP-9含量降低(P0.05)。3 RT-PCR和Western Blotting結果表明:隨SIN濃度升高,各處理組人胃癌SGC-7901細胞中MMP-2 m RNA、蛋白表達以及MMP-9 mRNA表達豐度均顯著降低(P0.05)。其中0.5mmol/L、1 mmol/L和2 mmol/L三個處理組MMP-9蛋白表達也顯著降低(P0.05);與對照組相比,0.5mmol/L、1mmol/L和2 mmol/L處理組Snail基因和蛋白表達顯著降低(P0.05);與對照組相比,0.5 mmol/L、1mmol/L和2 mmol/L處理組E-cadherin基因和蛋白表達顯著升高(P0.05)。4 Western Blotting結果發(fā)現(xiàn),隨著SIN濃度的升高,SGC-7901細胞核中NFκB p65蛋白表達呈現(xiàn)降低趨勢。與對照組相比,與0.5 mmol/L、1 mmol/L和2 mmol/L處理組均顯著降低(P0.05)。結論:1 SIN能抑制胃癌細胞SGC-7901的增殖,且具有時間和濃度依賴性;2 SIN對胃癌細胞SGC-7901中EMT相關蛋白表達和NFκB p65的核轉(zhuǎn)位具有一定的調(diào)節(jié)作用;3 NFκB p65可能通過核轉(zhuǎn)位來參與SN對SGC-7901的EMT調(diào)控作用。
[Abstract]:Aim: to study the effects of sinomenine on SGC-7901 proliferation, invasion and metastasis related genes and protein expression in human gastric cancer cells.Methods the survival rate of human gastric cancer SGC-7901 cells was detected by using different concentrations of SIN.WST-1 assay was used to detect the cell survival rate of 2. 2 MMP expression in human gastric cancer cells treated with different concentrations of SIN. Elisa was used to detect the changes of MMP-2 and MMP-9 content in cell culture medium. 3. 3 RT-PCR detection of SIN in different concentrations of SIN on human gastric cancer SGC-7901 cells.Effect of 4. 4 Western blotting on the expression of EMT protein in SGC-7901 cells of gastric carcinoma; the effect of SIN at different concentrations on the expression of NF 魏 B p65 protein in SGC-7901 cell nucleus of human gastric cancer.Results Human gastric cancer SGC-7901 cells were treated with different concentrations of SIN. The results showed that the survival rate of SGC-7901 cells treated with 1 mmol / L ~ (2 mmol / L) ~ (2.5 mmol/L) and 3mmol/L was significantly lower than that of control group (P _ (0.05)) after 24 h treatment.After 48 h treatment, the cell survival rate of 0.5 mmol / L ~ (-1) mmol / L ~ (2 mmol / L) 2 mmol / L ~ (2 mmol/L) and 3 mmol/L treatment group was significantly lower than that of the control group (P 0.05), and the MMPs content of SGC-7901 cell culture medium was decreased after treatment with sinomenine in a time-and dose-dependent manner.The content of MMP-2 in the culture medium of each SIN treatment group was significantly lower than that of the control group (P0.05T-1 mmol/L and 2 mmol/L). The results showed that the content of MMP-9 decreased with the increase of SIN concentration.The expression of MMP-2 m RNA, protein and MMP-9 mRNA in human gastric cancer SGC-7901 cells were significantly decreased in each treatment group.The expression of MMP-9 protein in 0.5 mmol / L mmol/L and 2 mmol/L groups also decreased significantly compared with the control group, and the Snail gene and protein expression in 0.5 mmol / L 1 mmol / L and 2 mmol/L treatment group was significantly lower than that in the control group, and the E-cadherin gene and protein surface in the 0. 5 mmol / L 1 mmol / L and 2 mmol/L treatment group was significantly lower than that in the control group.The results of P0.05n.4 Western Blotting showed that,The expression of NF 魏 B p65 in SGC-7901 cells decreased with the increase of SIN concentration.Compared with the control group, 0. 5 mmol 路L ~ (-1) L ~ (-1) mmol/L and 2 mmol/L treatment group significantly decreased P 0. 05%.Conclusion 1 SIN can inhibit the proliferation of SGC-7901 in gastric cancer cells.The expression of EMT related protein in SGC-7901 and nuclear translocation of NF 魏 B p65 may be involved in the EMT regulation of SGC-7901 by nuclear translocation of NF 魏 B p65 in gastric cancer cells.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R735.2

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本文編號：1728493