【摘要】：目的:探討活性氧(ROS)對卵巢上皮性癌細胞株SKOV3細胞的間充質轉化的影響及其中可能存在的分子機制。方法:1通過采用ROS生成劑大黃素(Emodin)、ROS清除劑二硫蘇糖醇(DTT)調節(jié)卵巢上皮性癌細胞株SKOV3細胞內ROS水平,采用PCR技術及蛋白印跡法測定3組(Emodin組、DTT組、對照組)細胞中間充質轉化標志分子E鈣黏蛋白(E-cadherin)mRNA及蛋白的表達,比較細胞株侵襲轉移能力。2結合改變活性氧水平,構建缺氧誘導因子-1α(HIF-1α)的化學合成,小干擾RNA(SiRNA)沉默HIF-1α表達(HIF-1αSiRNA組、DTT組、對照組),比較HIF-1αmRNA的表達。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,觀察各組(Emodin組、DTT組、β-APN組、Emodin+β-APN組、對照組)細胞HIF-1α、LOX、E-cadherin mRNA及蛋白表達。結果:1SKOV3細胞Emodin組ROS水平較對照組明顯升高[(164.85±8.93)%vs(97.42±6.47)%,P0.01],E-cadherin mRNA及蛋白的表達均較對照組顯著降低(P0.05),Emodin組的細胞侵襲率高于對照組[(19.48±0.49)%vs(10.74±2.38)%,P0.01],而DTT組結果與之相反,與對照組比較差異有統(tǒng)計學意義(P0.05)。2HIF-1αSiRNA作用下,HIF-1αSiRNA組的HIF-1αmRNA表達較對照組降低(0.20±0.09 vs 0.87±0.10,P0.05),LOX mRNA表達較對照組降低(0.40±0.08 vs 3.76±0.66,P0.05),而E-cadherin mRNA表達較對照組增高(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)計學意義(P0.05)。3β-APN作用下,β-APN組的HIF-1α蛋白表達較對照組無明顯變化(0.92±0.19 vs 0.94±0.09,P0.05),LOX蛋白表達較對照組降低(0.35±0.11 vs 0.81±0.13,P0.05),而E-cadherin蛋白表達較對照組增高(4.65±0.90 vs 2.23±0.61,P0.05)。結論:使ROS升高,上調HIF-1α,使LOX過表達,進而下調E-cadherin,ROS可能介導卵巢癌細胞間充質轉化,繼而發(fā)生侵襲轉移。
[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】： 上海交通大學醫(yī)學院附屬仁濟醫(yī)院;上海交通大學醫(yī)學院基礎醫(yī)學院細胞生物學教研室;
【基金】：上海市衛(wèi)生局科研項目(編號:2010272)
【分類號】：R737.31

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