本文關(guān)鍵詞： 滴滴涕 大腸癌 上皮間充質(zhì)轉(zhuǎn)化 信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子　出處：《中國藥理學(xué)與毒理學(xué)雜志》2017年02期 　論文類型：期刊論文

【摘要】：目的探究滴滴涕(DDT)對人結(jié)直腸腺癌上皮細(xì)胞(DLD1)上皮間充質(zhì)轉(zhuǎn)化的影響及機(jī)制。方法DLD1細(xì)胞用DDT 0.01,0.1,1.0,10.0和100.0 nmol·L~(-1)處理48 h后,倒置顯微鏡下觀察細(xì)胞形態(tài);實(shí)時(shí)熒光定量PCR法檢測E-鈣黏著蛋白、N-鈣黏著蛋白、波形蛋白和鋅指轉(zhuǎn)錄因子Snail1的mRNA表達(dá)。Western蛋白質(zhì)印跡法檢測信號轉(zhuǎn)導(dǎo)和轉(zhuǎn)錄激活因子3(STAT3)信號通路主要蛋白STAT3和p-STAT3的蛋白水平。用STAT3抑制劑WP1066(5μmol·L~(-1))處理,通過Western印跡法和實(shí)時(shí)熒光定量PCR法檢測其對DDT誘導(dǎo)的STAT3/Snail1信號通路中p-STAT3、STAT3的蛋白水平和上皮間充質(zhì)轉(zhuǎn)化關(guān)鍵因子E-鈣黏著蛋白、N-鈣黏著蛋白、波形蛋白和鋅指轉(zhuǎn)錄因子Snail1的mRNA水平的影響。結(jié)果與正常對照組相比,DLD1細(xì)胞在DDT處理48 h后,細(xì)胞形態(tài)由卵圓形逐漸變?yōu)殚L梭形,E-鈣黏著蛋白mRNA相對表達(dá)顯著降低(P0.01),為正常對照組的(42.4±2.8)%。N-鈣黏著蛋白和波形蛋白mRNA相對表達(dá)顯著提高(P0.01),為正常對照組的1.91±0.1倍和(1.5±0.2)倍。STAT3信號通路蛋白STAT3和p-STAT3蛋白表達(dá)均升高(P0.01),為正常對照組的2.1和1.8倍。鋅指轉(zhuǎn)錄因子Snail1的mRNA相對表達(dá)顯著升高(P0.01),是正常對照組的(1.5±0.1)倍。STAT3抑制劑WP1066 5μmol·L~(-1)處理后,鋅指轉(zhuǎn)錄因子Snail1 mRNA的表達(dá)明顯下調(diào)(P0.01),為DDT 1.0 nmol·L~(-1)處理組的(56.3±0.9)%,同時(shí)抑制DDT誘導(dǎo)的E-鈣黏著蛋白mRNA表達(dá)升高(P0.01),為DDT 1.0 nmol·L~(-1)處理組的2.5±0.1倍,N-鈣黏著蛋白和波形蛋白mRNA表達(dá)降低(P0.01),分別為DDT 1.0 nmol·L~(-1)處理組的(50.2±2.9)%和(61.6±6.1)%。結(jié)論 DDT可能通過STAT3/Snail1信號通路改變上皮間充質(zhì)轉(zhuǎn)化子E-鈣黏著蛋白、N-鈣黏著蛋白和波形蛋白的表達(dá),進(jìn)而促進(jìn)大腸癌細(xì)胞上皮間充質(zhì)轉(zhuǎn)化。
[Abstract]:Objective to investigate the effect and mechanism of DDT on the epithelial mesenchymal transformation of human colorectal adenocarcinoma cell line DLD1.Methods DLD1 cells were treated with DDT 0.01C 1.0 and 100.0 nmol 路L ~ (-1) for 48 h, and the morphology of the cells was observed under inverted microscope. Real time fluorescence quantitative PCR assay was used to detect E-cadherin (E-cadherin). MRNA expression of vimentin and zinc finger transcription factor Snail1. Western blot assay was used to detect the protein levels of STAT3 and p-STAT3 in signal transduction and transcription activator 3- STAT3 signaling pathway. STAT3 inhibitor WP1066(5 渭 mol. The protein level of p-STAT3- STAT3 in STAT3/Snail1 signaling pathway induced by DDT and the key factor of epithelial mesenchymal transformation, E-cadherin, were detected by Western blot and real-time fluorescence quantitative PCR. The effects of vimentin and zinc finger transcription factor (Snail1) on mRNA levels were observed. Results compared with the control group, DLD1 cells were treated with DDT for 48 h. The relative expression of E-cadherin mRNA decreased significantly from oval to fusiform, which was significantly increased by 1.91 鹵0.1 times and 1.5 鹵0.2 times of the normal control group (P = 42.4 鹵2.8) and vimentin mRNA (P < 0.01). The expression of STAT3 and p-STAT3 protein were increased by 2.1 and 1.8 times as compared with those of the normal control group. The mRNA expression of zinc finger transcription factor Snail1 was significantly higher than that of the normal control group, and was 1.5 鹵0.1 times higher than that of the normal control group. After treatment with WP1066 5 渭 mol 路L ~ (-1), a STAT3 inhibitor, the relative expression of zinc finger transcription factor Snail1 was significantly higher than that of the control group. The expression of zinc finger transcription factor Snail1 mRNA was significantly down-regulated in the DDT 1.0 nmol 路L ~ (-1) group, which was 56.3 鹵0.9% of the DDT 1.0 nmol 路L ~ (-1) group. At the same time, it inhibited the increase of DDT induced mRNA expression of E-cadherin, which was 2.5 鹵0.1 times lower than that of DDT 1.0 nmol 路L ~ (-1) group. Conclusion DDT may change the expression of E-cadherin and vimentin by STAT3/Snail1 signaling pathway. And then promote the epithelial mesenchymal transformation of colorectal cancer cells.
【作者單位】： 山西大學(xué)生物技術(shù)研究所化學(xué)生物學(xué)與分子工程教育部重點(diǎn)實(shí)驗(yàn)室;山西醫(yī)科大學(xué)第一醫(yī)院;
【基金】：國家自然科學(xué)基金(21207084);國家自然科學(xué)基金(31271516) 山西省自然科學(xué)基金(2014011027-5) 高等學(xué)校科技創(chuàng)新項(xiàng)目(2016122) 山西省回國留學(xué)人員科研資助項(xiàng)目(2016-115)~~
【分類號】：R735.34
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本文編號：1532786