LncRNA MALAT1对人脐静脉内皮细胞HUVEC增殖与凋亡影响

Effect of LncRNA MALAT1 on proliferation and apoptosis of human umbilical vein endothelial cells

目的探讨长链非编码RNA(lncRNA)肺腺癌相关转录本1(MALAT1)对人脐静脉内皮细胞HUVEC增殖和凋亡的影响。方法构建MALAT1-RNAi质粒载体,应用脂质体法转染人脐静脉内皮细胞HUVEC细胞;设空白细胞对照组、阴性对照组和MALAT1-RNAi转染组,采用实时荧光定量PCR(qRT-PCR)法检测各实验组细胞中MALAT1的表达水平;CCK8法检测细胞的增殖情况,流式细胞术Annexin V-FITC/PI双染检测各组细胞的凋亡能力。结果 qRT-PCR检测结果显示,MALAT1-RNAi转染组人脐静脉内皮细胞HUVEC中MALAT1的相对表达量为(0.051±0.03),均低于空白细胞对照组和阴性对照组(均P<0.01);CCK8检测结果显示,MALAT1-RNAi转染组培养48和72h的吸光度(A)值分别为(0.072±0.087)和(1.115±0.077),均低于空白细胞对照组和阴性对照组(均P<0.05);AnnexinⅤ-FITC/PI双染流式细胞术检测结果显示,MALAT1-RNAi转染组HUVEC细胞凋亡率为(14.08±1.05)%,均高于空白细胞对照组和阴性对照组(均P<0.01)。结论 MALAT1-RNAi可抑制人脐静脉内皮细胞HUVEC的增殖能力,并促进其凋亡。

Objective To study the effect of long non-coding RNA （LncRNA） metastasis-associated lung adenocarcinoma transcript 1 （MALAT1） on the proliferation and apoptosis of human umbilical vein endothelial cells （HUVECs）. Methods We constructed RNA interference （RNAi） plasmid vectors for MALAT1 and then transfected them into HUVECs by liposome method. A blank control and a negative control group of HUVECs were also established. The expression of MALAT1 in HUVECs of the three groups was detected with quantitative real-time PCR （ qRT-PCR）. The proliferation of HUVECs was determined with caerulein and cholecystokinin octapeptide （CCK-8） assay and the apoptosis of the HUVECs was analyzed by flow cytometry with annexin V-fluorescein isothiocyanate/propidium iodide （FITC/PI） double staining. Results The relative MALAT1 expression was 0.051 ±0.03 in MALAT1-RNAi transfected HUVECs and significantly higher than that of in blank and negative control HUVECs （both P 〈 0. 01 ） ; the absorbance values （A） of culture medium for MALAT1-RNAi transfeced HUVECs were 0. 072 ± 0. 087 and 1. 115 ± 0. 077 at 48 and 72 hours of incubation and were both significantly lower than those of the blank and negative control HUVECs （ P 〈 0. 05 for all） ;the apoptosis rate of MALAT1-RNAi transfeced HUVECs was 14. 08 ± 1.05% and significantly higher than that blank and negative control HUVECs （ both P 〈 0. 01 ）. Conclusion MALAT1 -RNAi could inhibit the proliferation and promote the apoptosis of human umbilical vein endothelial cells.