發(fā)布時間：2018-06-28 21:00

  本文選題：人臍靜脈血管內(nèi)皮細胞 + 胞質(zhì)多聚腺苷酸結合蛋白　； 參考：《山東醫(yī)藥》2017年09期

【摘要】：目的探討胞質(zhì)多聚腺苷酸結合蛋白1(CPEB1)對人臍靜脈血管內(nèi)皮細胞(HUVECs)增殖和遷移的影響及可能機制。方法取對數(shù)生長期的HUVECs細胞,分為實驗組與對照組,分別用腫瘤細胞上清液培養(yǎng)基和普通培養(yǎng)基重懸細胞,Real-time PCR法和Western blotting法分別檢測CPEB1及整合素-金屬蛋白酶17(ADAM17)的mRNA和蛋白表達;取對數(shù)生長期HUVECs細胞,待培養(yǎng)至細胞達70%~80%融合時用LipofectamineTM2000進行轉染,細胞分為攜帶基因的質(zhì)粒轉染組(轉染組)、不帶任何基因的空質(zhì)粒轉染細胞組(轉染對照組)和不經(jīng)任何處理的細胞組(空白對照組)。MTT法檢測轉染后HUVECs的增殖能力,Transwell小室法檢測轉染后HUVECs的遷移能力,RT-PCR法檢測ADAM17的mRNA表達,Western blotting法檢測ADAM17蛋白表達。結果實驗組中CPEB1的mRNA及蛋白表達均低于對照組(P均0.05),ADAM17的mRNA及蛋白表達均高于對照組(P均0.05);轉染48、72 h后,轉染組吸光度值低于轉染對照組和空白對照組(P均0.05);轉染組細胞穿過小室膜數(shù)目低于轉染對照組和空白對照組(P均0.05);轉染組ADAMl7的mRNA和蛋白表達低于轉染對照組和空白對照組(P均0.05)。結論 CPEB1可抑制HUVECs細胞增殖和遷移,其機制可能與調(diào)控ADAM17表達有關。
[Abstract]:Objective to investigate the effect of cytosolic polyadenylate binding protein-1 (CPEB1) on proliferation and migration of human umbilical vein endothelial cells (HUVECs) and its possible mechanism. Methods HUVECs cells in logarithmic growth stage were divided into experimental group and control group. The mRNA and protein expression of CPEB1 and integrin-metalloproteinase-17 (ADAM17) were detected by real-time PCR and Western blotting in tumor cell supernatant medium and ordinary culture medium respectively. HUVECs cells in logarithmic growth phase were transfected with Lipofectamine TM2000 when the cells reached 70% fusion. Cells were divided into plasmid transfection group carrying gene (transfection group), empty plasmid transfection group without any gene (transfection control group) and cell group without any treatment (blank control group). MTT assay was used to detect the proliferation of HUVECs after transfection. MRNA expression of ADAM17 was detected by RT-PCR and the expression of ADAM17 protein was detected by Western blotting. Results the expression of CPEB1 mRNA and protein in the experimental group was lower than that in the control group (P 0.05). The absorbance value of transfection group was lower than that of transfection control group and blank control group (P 0.05), the number of cells passing through ventricular membrane in transfection group was lower than that in transfection control group and blank control group (P 0.05), the mRNA and protein expression of ADAMl7 in transfection group was lower than that in transfection control group. Blank control group (P 0.05). Conclusion CPEB1 can inhibit the proliferation and migration of HUVECs cells, and its mechanism may be related to the regulation of ADAM17 expression.
【作者單位】： 華中科技大學同濟醫(yī)學院附屬武漢中心醫(yī)院;華中科技大學同濟醫(yī)學院附屬同濟醫(yī)院;
【基金】：華中科技大學同濟醫(yī)學院附屬武漢中心醫(yī)院院內(nèi)科研基金項目(YQ14A03)
【分類號】：R730.2
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本文編號：2079336