环状RNA circ0004771对肺腺癌细胞A549的增殖、迁移与侵袭的影响

目的 探讨环状RNA circ0004771对肺腺癌细胞增殖、迁移与侵袭的影响及其机制。方法 培养肺腺癌A549细胞,分为阴性对照（si-NC）组及circ0004771小干扰RNA（siRNA）组（si-circ0004771组）,将si-NC和circ0004771 siRNA分别转染入相应组A549细胞中。si-NC组和si-circ0004771组细胞转染后,通过实时荧光定量聚合酶链反应检测2组细胞circ0004771、miR-339-5p的表达水平,通过细胞计数试剂盒法检测细胞活力,通过EdU实验检测2组细胞增殖能力,通过Transwell小室实验检测2组细胞迁移和侵袭能力。结果 si-NC组和si-circ0004771组A549细胞circ0004771基因的相对表达水平分别为3.07±0.07和0.68±0.04,miR-339-5p相对表达水平分别为1.14±0.13和2.33±0.07,差异均有统计学意义（t=51.34、13.96,P值均<0.001）。si-NC组和si-circ0004771组A549细胞在培养前（0 h）和培养后24、48 h的吸光度值分别为0.21±0.02、0.35±0.05、0.65±0.04和0.21±0.01、0.33±0.04、0.59±0.05,差异均无统计学意义（P值均>0.05）;而在培养后72 h si-circ0004771组吸光度为0.92±0.15, 小于si-NC组的1.32±0.04,差异有统计学意义（t=4.46,P=0.011）。EdU实验中,si-circ0004771组细胞阳性率为47.97%±4.68%,低于si-NC组的58.61%±2.15%,差异有统计学意义（t=3.58,P=0.023）。转染后si-circ0004771组的细胞迁移率、侵袭率分别为52.27%±2.92%、55.33%±6.29%,均低于si-NC组的99.79%±4.23%、98.67%±5.72%,差异均有统计学意义（t=16.26、8.83,P值均<0.001）。结论 下调肺腺癌A549细胞circ0004771的表达水平,可以降低A549细胞的活力和增殖能力,可抑制A549细胞的迁移、侵袭能力,其作用机制可能与上调miR-339-5p的表达有关。

Effects of circRNA circ0004771 on the proliferation,migration, and invasion of lung adenocarcinoma cell line A549

Objective To investigate the biological role of circ0004771 in lung adenocarcinoma. Methods Lung adenocarcinoma A549 cells were cultured and divided into the negative control (si-NC) and circ0004771 siRNA (si-circ0004771) groups. Si-NC and circ0004771 siRNA were transfected into A549 cells in the corresponding groups. After the transfection of the cells in the si-NC and si-circ0004771 groups, the expression levels of circ0004771 and miR-339-5p were detected via real-time fluorescent quantitative polymerase chain reaction; cell viability was determined through cell counting kit-8; cell proliferation was detected by using the EdU assay; and cell migration and invasion were detected through the Transwell chamber assay. Results In the A549 cells in the si-NC and si-circ0004771 groups, the relative expression levels of the circ0004771 gene were 3.07±0.07 and 0.68±0.04, respectively, and the relative expression levels of miR-339-5p were 1.14±0.13 and 2.33±0.07, respectively; these differences were statistically significant (t=51.34,13.96, all P values < 0.001). The absorbance values of the A549 cells in the si-NC and si-circ0004771 groups at 0, 24, and 48 h were 0.21±0.02, 0.35±0.05, and 0.65±0.04, respectively, and 0.21±0.01,0.33±0.04, and 0.59±0.05, respectively; no significant difference was observed (all P values > 0.05). However, at 72 h, the absorbance of the si-circ0004771 group was 0.92±0.15, which was less than that of the si-NC group (1.32±0.04); this difference was statistically significant (t=4.46, P=0.011). In the EdU experiment, the positive rate of cells in the si-circ0004771 group was 47.97%±4.68%, which was lower than the positive rate of 58.61%±2.15% shown by the si-NC group; this difference was statistically significant (t=3.58, P=0.023). After transfection, the cell migration and invasion rates in the si-circ0004771 group were 52.27%±2.92% and 55.33%±6.29%, respectively, and were all lower than the cell migration and invasion rates of 99.79%±4.23% and 98.67% ±5.72% in the si-NC group; this difference was statistically significant (t=16.26, 8.83, all P values < 0.001). Conclusions Down regulating the expression level of circ0004771 in lung adenocarcinoma cell line A549 can reduce the viability and proliferation of A549 cells and inhibit the migration and invasion of A549 cells; the mechanism of this effect may be related to the upregulation of mir-339-5p expression.