【摘要】：目的:探討活性氧(ROS)對(duì)卵巢上皮性癌細(xì)胞株SKOV3細(xì)胞的間充質(zhì)轉(zhuǎn)化的影響及其中可能存在的分子機(jī)制。方法:1通過(guò)采用ROS生成劑大黃素(Emodin)、ROS清除劑二硫蘇糖醇(DTT)調(diào)節(jié)卵巢上皮性癌細(xì)胞株SKOV3細(xì)胞內(nèi)ROS水平,采用PCR技術(shù)及蛋白印跡法測(cè)定3組(Emodin組、DTT組、對(duì)照組)細(xì)胞中間充質(zhì)轉(zhuǎn)化標(biāo)志分子E鈣黏蛋白(E-cadherin)mRNA及蛋白的表達(dá),比較細(xì)胞株侵襲轉(zhuǎn)移能力。2結(jié)合改變活性氧水平,構(gòu)建缺氧誘導(dǎo)因子-1α(HIF-1α)的化學(xué)合成,小干擾RNA(SiRNA)沉默HIF-1α表達(dá)(HIF-1αSiRNA組、DTT組、對(duì)照組),比較HIF-1αmRNA的表達(dá)。3使用賴氨酰氧化酶(LOX)抑制劑β-氨基丙腈(β-APN)抑制LOX表達(dá),觀察各組(Emodin組、DTT組、β-APN組、Emodin+β-APN組、對(duì)照組)細(xì)胞HIF-1α、LOX、E-cadherin mRNA及蛋白表達(dá)。結(jié)果:1SKOV3細(xì)胞Emodin組ROS水平較對(duì)照組明顯升高[(164.85±8.93)%vs(97.42±6.47)%,P0.01],E-cadherin mRNA及蛋白的表達(dá)均較對(duì)照組顯著降低(P0.05),Emodin組的細(xì)胞侵襲率高于對(duì)照組[(19.48±0.49)%vs(10.74±2.38)%,P0.01],而DTT組結(jié)果與之相反,與對(duì)照組比較差異有統(tǒng)計(jì)學(xué)意義(P0.05)。2HIF-1αSiRNA作用下,HIF-1αSiRNA組的HIF-1αmRNA表達(dá)較對(duì)照組降低(0.20±0.09 vs 0.87±0.10,P0.05),LOX mRNA表達(dá)較對(duì)照組降低(0.40±0.08 vs 3.76±0.66,P0.05),而E-cadherin mRNA表達(dá)較對(duì)照組增高(1.01±0.15 vs 0.30±0.09,P0.05)。DTT組的效果較HIF-1αSiRNA組更為顯著,兩組比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。3β-APN作用下,β-APN組的HIF-1α蛋白表達(dá)較對(duì)照組無(wú)明顯變化(0.92±0.19 vs 0.94±0.09,P0.05),LOX蛋白表達(dá)較對(duì)照組降低(0.35±0.11 vs 0.81±0.13,P0.05),而E-cadherin蛋白表達(dá)較對(duì)照組增高(4.65±0.90 vs 2.23±0.61,P0.05)。結(jié)論:使ROS升高,上調(diào)HIF-1α,使LOX過(guò)表達(dá),進(jìn)而下調(diào)E-cadherin,ROS可能介導(dǎo)卵巢癌細(xì)胞間充質(zhì)轉(zhuǎn)化,繼而發(fā)生侵襲轉(zhuǎn)移。
[Abstract]:Objective: To study the effect of reactive oxygen (ROS) on the mesenchymal transition of ovarian epithelial cell line SKOV3 cells and the possible molecular mechanism of ROS. Methods:1. The ROS level in the SKOV3 cells of the epithelial ovarian cancer cell line SKOV3 was adjusted by using the ROS-generating agent Emodin and the ROS-scavenger dithiothreitol (DTT), and the three groups (Emodin group and DTT group) were determined by PCR and Western blot. In the control group, the expression of E-cadherin (E-cadherin) mRNA and protein was compared with the expression of E-cadherin (E-cadherin) mRNA and protein, and the invasion and metastasis ability of the cell line was compared. The expression of HIF-1 mRNA was compared with the expression of HIF-1. The expression of HIF-1, LOX, E-cadherin and the expression of protein in each group (Emodin group, DTT group, P-APN group, Emodin + 1-APN group and control group) were observed. Results: The level of ROS in the Emodin group of 1SKOV3 cells was significantly higher than that in the control group[(164.85-8.93)% vs (97.42-6.47)%, and the expression of E-cadherin mRNA and protein in the Emodin group was significantly lower than that in the control group (P0.05), and the cell invasion rate of the Emodin group was higher than that of the control group[(19.48-0.49)% vs (10.74-2.38)%, P0.01], Compared with the control group, the expression of HIF-1 mRNA in the HIF-1-SiRNA group was lower than that in the control group (0.20-0.09 vs 0.87-0.10, P0.05), while the expression of LOX mRNA was lower in the control group (0.40% 0.08 vs 3.76-0.66, P0.05). The expression of E-cadherin mRNA was higher in the control group (1.01-0.15 vs 0.30-0.09, P0.05). The effect of the DTT group was more significant than that of the HIF-1/ SiRNA group. The difference between the two groups was statistically significant (P <0.05). The expression of HIF-1 in the P-APN group was not significantly changed in the control group (0.92, 0.19 vs. 0.94, 0.09, P0.05), and the expression of LOX protein was lower in the control group (0.35-0.11 vs. 0.81-0.13, The expression of E-cadherin was higher in the control group (4.65, 0.90 vs 2.23, 0.61, P0.05). Conclusion: The increase of ROS, the up-regulation of HIF-1, the overexpression of LOX, and the further down-regulation of E-cadherin, ROS may mediate the transformation of human ovarian cancer cells, and then the invasion and metastasis.
【作者單位】： 上海交通大學(xué)醫(yī)學(xué)院附屬仁濟(jì)醫(yī)院;上海交通大學(xué)醫(yī)學(xué)院基礎(chǔ)醫(yī)學(xué)院細(xì)胞生物學(xué)教研室;
【基金】：上海市衛(wèi)生局科研項(xiàng)目(編號(hào):2010272)
【分類號(hào)】：R737.31

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 盧玉娟;羅yN;楊雅瑩;易永芬;;高爾基體α-甘露糖苷酶Ⅱ、E-cadherin和α-catenin在卵巢上皮腫瘤組織和不同轉(zhuǎn)移能力的卵巢癌細(xì)胞中的表達(dá)[J];腫瘤;2012年11期

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張建蓉;佘強(qiáng);;上皮-間質(zhì)轉(zhuǎn)化在心臟發(fā)育和損傷中的作用[J];國(guó)際心血管病雜志;2013年05期

2 Xiang-Ming Ding;;MicroRNAs: regulators of cancer metastasis and epithelialmesenchymal transition(EMT)[J];Chinese Journal of Cancer;2014年03期

3 王一;黎燕;沈倍奮;陳國(guó)江;;炎癥:腫瘤的推進(jìn)器[J];國(guó)際藥學(xué)研究雜志;2014年01期

4 陳杰;郭維華;田衛(wèi)東;;牙囊和上皮根鞘細(xì)胞成無(wú)細(xì)胞牙骨質(zhì)的機(jī)制[J];國(guó)際口腔醫(yī)學(xué)雜志;2014年03期

5 章俊;邱敏姿;馬亞瓊;卜陽(yáng);楊蕾;湯s，

本文編號(hào)：2512290