miR-21在脂多糖诱导的人鼻黏膜上皮细胞中表达变化及作用机制

目的探讨微RNA-21(miR-21)在脂多糖诱导的人鼻黏膜上皮细胞(HNEpC)中表达变化及可能的作用机制。方法应用脂多糖诱导建立HNEpC炎症反应细胞模型,分别应用荧光定量PCR(qRT-PCR)和蛋白免疫印迹法检测炎症细胞模型建立前后miR-21和磷酸酶及张力蛋白同源物(PTEN)蛋白的表达水平变化。取脂多糖诱导的HNEpC细胞,分为miR-21抑制物组和阴性对照组,应用脂质体转染法分别将miR-21抑制物和阴性对照物转染入2组HNEpC细胞,比较转染后2组细胞中miR-21和PTEN蛋白的表达、细胞增殖活性和凋亡率以及细胞上清液中TNF-α、IL-1β、IL-6和IL-10的水平。结果脂多糖诱导的HNEpC细胞中miR-21表达水平增高(P <0.05),PTEN蛋白的表达水平降低(P <0.05)。miR-21抑制物组细胞中miR-21表达水平低于阴性对照组(P <0.05),转染后24、48、72 h后的吸光度值均低于阴性对照组(P均<0.05),凋亡率高于阴性对照组(P <0.05),并且细胞上清液中TNF-α、IL-1β及IL-6水平均低于...

Changes of miR-21 expression and mechanism in human nasal epithelial cells induced by lipopo-lysaccharide

Objective To investigate the changes of microRNA-21 （miR-21） expression in human nasal epithelial cells （HNEpC） induced by lipopolysaccharide （LPS） and unravel the potential mechanism. Methods HNEpC inflammatory cell model was induced by LPS. The changes of the expression levels of miR-21 and phosphatase and tensin homologue （PTEN） protein before and after establishment of the inflammatory cell model were detected by qRT-PCR and Western blot. HNEpC cells induced by LPS were collected and divided into the miR-21 inhibitor group and negative control group. HNEpC cells were transfected with miR-21 inhibitor and negative controls by using the liposome transfection method. After cell transfection, the expression levels of miR-21 and PTEN protein, cell proliferation activity, apoptosis rate, and the expression levels of TNF-α, IL-1β, IL-6 and IL-10 in the cell supernatant were measured and statistically compared between two groups. Results In the HNEpC cells induced by LPS, the expression level of miR-21 was significantly up-regulated （P < 0.05）, whereas that of PTEN protein was significantly down-regulated （P < 0.05）. Compared with the negative control group, the expression level of miR-21 was significantly lower （P < 0.05）, the OD values at 24, 48 and 72 h after cell transfection were significantly lower （all P < 0.05）, the apoptosis rate was significantly higher （P < 0.05）, the expression levels of TNF-α, IL-1β and IL-6 in the cell supernatant were significantly lower （all P < 0.05）, the expression level of IL-10 was significantly higher （P < 0.05）,and the expression level of PTEN protein was significantly higher in the miR-21 inhibitor group （P < 0.05）, respectively. Conclusions The expression level of miR-21 is up-regulated in the LPS-induced HNEpC inflammatory cell model. Inhibition of miR-21 expression can significantly suppress the proliferation activity and inflammatory response of HNEpC cells induced by LPS and promote the cell apoptosis. The underlying mechanism may be related to the regulation of PTEN gene.