长链非编码RNA UCA1对肺腺癌A549细胞糖代谢及侵袭转移的影响

The functional role of long non-coding RNA UCA1 in lung adenocarcinoma A549 cells on glucose metabolism,invasion and metastasis

目的探讨长链非编码RNA(lncRNA)尿路上皮癌胚抗原1(UCA1)在肺腺癌A549细胞中的表达及对糖代谢关键酶和细胞侵袭、转移的影响。方法构建lncRNA UCA1沉默及其对照稳定转染的肺腺癌A549细胞系,将小干扰RNA(siRNA)转染细胞分为NC组(阴性对照组,转染siRNA-UCA1序列)和siRNA-UCA1组(转染siRNA-UCA1敲降序列)。采用实时荧光定量PCR、Western blot检测糖代谢相关指标葡萄糖转运蛋白(GLUT)1、己糖激酶(HK)2和丙酮酸激酶(PKM)2水平的变化。采用Transwell实验和划痕实验检测siRNA-UCA1的侵袭和迁移能力。采用18F-氟脱氧葡萄糖(FDG)摄取实验检测siRNA-UCA1的摄取率。计量资料的比较采用t检验。结果UCA1在肺腺癌A549细胞中高表达。与NC组相比,siRNA-UCA1组能够抑制GLUT1、HK2和PKM2基因在肺腺癌A549细胞中的表达(t=19.66、5.81、11.98,均P<0.001),且三者的蛋白表达水平明显下降(t=61.35、145.90、88.19,均P<0.001)。与NC组相比,siRNA-UCA1组肺腺癌A549细胞的侵袭能力、迁移运动能力和对18F-FDG的摄取率均明显降低(t=19.43、7.71、5.79,均P<0.05)。结论LncRNA UCA1能够抑制糖代谢的关键酶并促进肺腺癌的转移能力,其可能成为肺癌新的诊断指标和治疗靶点。

Objective To explore the expression of long non-coding RNA(lncRNA)urothelial carcinoembryonic antigen 1(UCA1)in lung adenocarcinoma A549 cells and its influence on key enzymes involved in sugar metabolism,cell invasion,and metastasis.Methods The lncRNA UCA1 silencing and its control stable transfected lung adenocarcinoma A549 cells were constructed,the small interfering RNA(siRNA)transfected cells were divided into NC group(negative control group,transfected with siRNA-UCA1 sequence)and siRNA-UCA1 group(transfected with siRNAUCA1 knockdown sequence).Quantitative real-time PCR and Western blot analyses were used to detect changes in the levels of glucose transporter protein(GLUT)1,hexokinase(HK)2 and pyruvate kinase isozymes M(PKM)2 related to glucose metabolism.Transwell and scratch tests were conducted to detect the invasion and migration ability of siRNA-UCA1.18F-Fluorodeoxyglucose(FDG)uptake experiment was used to detect the uptake rate of siRNA-UCA1.T test was used to compare the measurement data.Results UCA1 was highly expressed in lung adenocarcinoma A549 cell line.Compared with the NC group,the siRNA-UCA1 group can inhibit the expression of GLUT1,HK2,and PKM2 in lung adenocarcinoma A549 cells(t=19.66,5.81,11.98;all P<0.001),and the protein expression levels of the three significantly decreased(t=61.35,145.90,88.19;all P<0.001).Compared with the NC group,the siRNA-UCA1 group's migration ability and 18F-FDG uptake rate of A549 cells were significantly reduced(t=19.43,7.71,5.79;all P<0.05).Conclusions LncRNA UCA1 can affected the key enzymes of glucose metabolism and promoted the metastasis ability of lung adenocarcinoma.Thus,lncRNA UCA1 may be a new diagnostic index and therapeutic target for lung cancer.