本文選題：PD-L　切入點(diǎn)：免疫治療　出處：《生物技術(shù)》2017年06期

【摘要】：[目的]探討PD-L1過表達(dá)對(duì)HeLa細(xì)胞遷移的影響。[方法]通過分子克隆構(gòu)建PD-L1質(zhì)粒載體(p EGFPPD-L1);通過PEI法轉(zhuǎn)染質(zhì)粒,調(diào)節(jié)質(zhì)粒轉(zhuǎn)染量(1μg、3μg、5μg)和轉(zhuǎn)染時(shí)間(24 h、36 h、48 h、72 h),優(yōu)化轉(zhuǎn)染條件;通過細(xì)胞劃痕實(shí)驗(yàn)檢測細(xì)胞融合率,Western Blot檢測細(xì)胞遷移相關(guān)蛋白(E-鈣黏蛋白、N-鈣黏蛋白、波形蛋白)的表達(dá)量。[結(jié)果]最佳轉(zhuǎn)染條件是質(zhì)粒量3μg/孔,轉(zhuǎn)染后48 h觀察,此時(shí)轉(zhuǎn)染效率可達(dá)(82.94±8.08)%。轉(zhuǎn)染pEGFP-PD-L1質(zhì)粒后,HeLa細(xì)胞的PD-L1過表達(dá)3.5倍,劃痕融合率顯著增高(p0.01);波形蛋白和N-鈣黏蛋白的表達(dá)量顯著升高(p0.05),E-鈣黏蛋白表達(dá)量顯著下降(p0.05)。[結(jié)論]PD-L1過表達(dá)顯著促進(jìn)HeLa細(xì)胞遷移能力和上皮向間質(zhì)轉(zhuǎn)化水平。
[Abstract]:[objective] to investigate the effect of overexpression of PD-L1 on the migration of HeLa cells. [methods] to construct PD-L1 plasmid vector pEGFPPD-L _ (1) by molecular cloning, to adjust the amount of 1 渭 g ~ (3 渭 g) ~ 5 渭 g ~ (5 渭 g) and the transfection time of 24 h ~ 36 h ~ 48 h ~ (72 h) to optimize the transfection conditions. The cell fusion rate and the expression of E-cadherin (vimentin) were detected by cell scratch assay and Western Blot. [results] the optimal transfection conditions were as follows: the amount of plasmid was 3 渭 g / well and 48 h after transfection, the expression of E-cadherin (vimentin) was detected. The transfection efficiency was 82.94 鹵8.08. The PD-L1 expression in HeLa cells was 3.5-fold after transfection of pEGFP-PD-L1 plasmid. The expression of vimentin and N- cadherin decreased significantly. [conclusion] overexpression of PD-L1 significantly promoted the migration ability of HeLa cells and the level of epithelium to mesenchymal transformation.
【作者單位】： 武漢科技大學(xué)生物醫(yī)學(xué)研究院生命科學(xué)與健康學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(“MRTF-A和STAT3調(diào)控乳腺癌EMT及機(jī)制研究”,No.31501149;“抗活性氧自由基的載白藜蘆醇Ti O2納米管促進(jìn)骨修復(fù)研究”,No.31700824)
【分類號(hào)】：R737.33

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