【摘要】：目的研究不同濃度水楊酸處理對(duì)胃癌細(xì)胞增殖和侵襲轉(zhuǎn)移能力的影響,并初步探討水楊酸抗腫瘤轉(zhuǎn)移的可能機(jī)制。方法采用一系列濃度梯度(0.5~3mmol/L)的水楊酸處理胃癌細(xì)胞株MGC-803,MTS比色法觀察不同濃度水楊酸處理24 h后細(xì)胞增殖能力的變化,篩選出最適的水楊酸處理濃度;細(xì)胞黏附實(shí)驗(yàn)觀察0.5mmol/L和1mmol/L水楊酸處理后細(xì)胞黏附能力的變化;Transwell觀察0.5mmol/L和1mmol/L水楊酸處理后細(xì)胞遷移和侵襲的能力變化;熒光定量PCR檢測(cè)0.5mmol/L和1mmol/L水楊酸處理后細(xì)胞黏附分子CD44、E鈣粘素(CDH1)、連環(huán)蛋白A1(CTNNA1)、纖連蛋白(FN1)、核纖層蛋白(Lamin)、核纖層蛋白B1(Lamin B1)、核纖層蛋白C1(Lamin C1)和整合素A9(ITGA9)m RNA表達(dá)水平的變化;Western-blot觀察細(xì)胞黏附分子CD44、CTNNA1、FN1以及Lamin B1蛋白表達(dá)水平的差異。結(jié)果MTS結(jié)果示0.5mmol/L和1mmol/L的水楊酸處理24h后細(xì)胞抑制率分別為(32±4.5)%和(38±4.1)%,低于50%,因此選取0.5mmol/L和1mmol/L的水楊酸作為后續(xù)實(shí)驗(yàn)的研究。細(xì)胞黏附實(shí)驗(yàn)顯示相比于對(duì)照組的細(xì)胞粘附數(shù)量(121.00±2.65)個(gè),0.5mmol/L組和1mmol/L組水楊酸處理后黏附的細(xì)胞數(shù)量明顯減少,分別是(88.00±1.53)個(gè)和(44.00±2.00)個(gè);細(xì)胞遷移實(shí)驗(yàn)示0.5mmol/L和1mmol/L組水楊酸處理后細(xì)胞遷移率明顯受到抑制,分別是(10.00±3.85)%和(12.00±3.61)%;細(xì)胞侵襲的結(jié)果示0.5mmol/L和1mmol/L水楊酸處理后,細(xì)胞OD值分別是(0.18±0.00)和(0.15±0.00),較正常組OD值(0.22±0.01)低。熒光定量PCR表明0.5mmol/L組和1mmol/L組水楊酸處理明顯下調(diào)細(xì)胞黏附分子CD44、CTNNA1、FN1、ITGA9、Lamin、Lamin B1和Lamin C1 m RNA水平的轉(zhuǎn)錄,上調(diào)CDH1 m RNA的表達(dá);Western-blot結(jié)果同樣證實(shí)0.5mmol/L和1mmol/L組水楊酸處理后細(xì)胞黏附分子CD44、CTNNA1、FN1和Lamin B1蛋白的表達(dá)水平明顯降低(P0.05)。結(jié)論1.0.5mmol/L和1mmol/L水楊酸可明顯抑制胃癌細(xì)胞株的體外增殖、黏附、遷移和侵襲,并且其抑制效應(yīng)隨著濃度的增加而逐漸增強(qiáng)。2.0.5mmol/L和1mmol/L水楊酸下調(diào)細(xì)胞黏附分子CD44、CTNNA1、FN1、ITGA9、Lamin、Lamin B1和Lamin C1的表達(dá)和上調(diào)CDH1的表達(dá),其抗腫瘤轉(zhuǎn)移的機(jī)制可能與抑制細(xì)胞黏附分子的表達(dá)和抑制上皮間質(zhì)轉(zhuǎn)化相關(guān)。
[Abstract]:Objective to study the effects of salicylic acid treatment on the proliferation, invasion and metastasis of gastric cancer cells, and to explore the possible mechanism of salicylic acid against tumor metastasis. Methods A series of concentration gradient (0.5~3mmol/L) salicylic acid treated gastric cancer cell line MGC-803,MTS colorimetry was used to observe the change of cell proliferation ability after 24 h treatment with different concentrations of salicylic acid, and the optimal concentration of salicylic acid was selected. Cell adhesion assay was used to observe the changes of cell adhesion ability after 0.5mmol/L and 1mmol/L salicylic acid treatment and Transwell to observe the ability of cell migration and invasion after 0.5mmol/L and 1mmol/L salicylic acid treatment. 0.5mmol/L and 1mmol/L salicylic acid treated cell adhesion molecule CD44,E cadherin (CDH1), catenin A1 (CTNNA1), fibronectin (FN1) and nuclear laminin (Lamin), (Lamin B1) were detected by fluorescence quantitative PCR. Changes of expression of nuclear laminin C1 (Lamin C1) and integrin A9 (ITGA9) m RNA); The expression levels of CD44,CTNNA1,FN1 and Lamin B1 were observed by Western-blot. Results MTS results showed that the inhibitory rates of 0.5mmol/L and 1mmol/L were (32 鹵4.5)% and (38 鹵4.1)% after 24 h treatment, respectively, which were lower than 50%. Therefore, the salicylic acid of 0.5mmol/L and 1mmol/L was selected as the follow-up study. Cell adhesion assay showed that compared with the control group (121.00 鹵2.65), the number of cell adhesion in 0.5mmol/L group and 1mmol/L group was significantly decreased after salicylic acid treatment. They were (88.00 鹵1.53) and (44.00 鹵2.00); Cell migration assay showed that the cell migration rate of 0.5mmol/L and 1mmol/L groups was (10.00 鹵3.85)% and (12.00 鹵3.61)%, respectively. The results of cell invasion showed that the OD values of cells treated with 0.5mmol/L and 1mmol/L salicylic acid were (0.18 鹵0.00) and (0.15 鹵0.00), respectively, which were lower than those of normal group (0.22 鹵0.01). Fluorescence quantitative PCR showed that salicylic acid treatment in 0.5mmol/L group and 1mmol/L group significantly down-regulated the transcription of cell adhesion molecules CD44,CTNNA1,FN1,ITGA9,Lamin,Lamin B1 and Lamin C 1m RNA, and up-regulated the expression of CDH1 m RNA. Western-blot results also confirmed that the expression levels of CD44,CTNNA1,FN1 and Lamin B1 were significantly decreased in 0.5mmol/L and 1mmol/L groups after salicylic acid treatment (P0.05). Conclusion 1.0.5mmol/L and 1mmol/L salicylic acid can significantly inhibit the proliferation, adhesion, migration and invasion of gastric cancer cell lines in vitro. 2.0.5mmol/L and 1mmol/L salicylic acid down-regulated the expression of CD44,CTNNA1,FN1,ITGA9,Lamin,Lamin B1 and Lamin C1 and up-regulated the expression of CDH1. The mechanism of its anti-tumor metastasis may be related to the inhibition of the expression of cell adhesion molecules and the inhibition of epithelial mesenchymal transformation.
【學(xué)位授予單位】：寧波大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

【參考文獻(xiàn)】

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1 李文;胡U，

本文編號(hào)：2404468