Hsacirc0002360在肺腺癌中的表达及临床意义

Expression of Hsacirc0002360 in lung adenocarcinoma and its clinical significance

目的目前环状RNA(circRNA)与肺腺癌的关系有待进一步探究。文中旨在探讨Hsa_circ_0002360在肺腺癌中的表达水平及其在肺腺癌临床诊断、治疗中的价值。方法收集2018年5月至2019年6月江苏大学附属人民医院胸外科107例手术切除的肺腺癌及癌旁组织标本。采集50例肺腺癌患者术前、术后及50例门诊体检健康者的外周血5 mL,分为肺腺癌组和正常对照组。采用qRT-PCR检测2组Hsa_circ_0002360在血液中的表达量。细胞培养人肺腺癌细胞株(A549、H1299、H1975)及人支气管黏膜上皮细胞株(BEAS-2B)。取6孔板分别接种H1975、A549肺腺癌细胞,每个板设置3个孔为小干扰RNA组(Si-circRNA),另3个孔为阴性对照组(Si-NC),进行细胞转染实验。采用CCK8法、细胞克隆形成实验、细胞划痕实验分别检测Hsa_circ_0002360对A549、H1975细胞活力、增殖能力及侵袭转移能力的影响。结果肺腺癌组织中hsacirc0002360的表达水平显著高于癌旁组织[(2.84±0.12)vs(1.46±0.08),P<0.01]。肺腺癌组患者血清中hsa_circ_0002360的表达水平显著高于正常对照组[(2.59±0.17)vs(1.21±0.11),P<0.01],肺腺癌患者术后血清中hsa_circ_0002360表达水平较术前明显下降[(1.58±0.09)vs(2.59±0.17),P<0.01]。与BEAS-2B hsa_circ_0002360的表达相比,A549、H1299、H1975肺腺癌细胞株中明显增加(P<0.05)。A549细胞中,小干扰RNA组细胞克隆形成率、细胞迁移率[(38.33±1.69)%、(13.5±3.34)%]明显低于阴性对照组[(65.66±4.93)%、(54.66±4.1)%],差异有统计学意义(P<0.01);H1975细胞亦明显低于阴性对照组(P<0.01)。结论Hsa_circ_0002360对肺腺癌早期诊断有评估价值,可作为肺腺癌早期诊断及治疗的生物标志物。

Objective The relationship between circular RNAs(circRNA)and lung adenocarcinoma(LAC)remains to be further explored.This study aimed to investigate the expression of Hsa_circ_0002360 in LAC and its value in the clinical diagnosis and Methods This study included 50 cases of LAC treated in our hospital from May 2018 to June 2019 and another 50 healthy subjects as normal controls.We determined the expression of Hsa_circ_0002360 in the LAC cells by qRT-PCR.We cultured human LAC cell lines A549,H1299 and H1975 and human bronchial mucosal epithelial cell line BEAS-2 B,inoculated the A549 and H1975 LAC cells in the 6-well plate,with 3 wells for si-circRNAs and the other 3 for negative controls(si-NC),followed by cell transfection experiment.Using the CCK8,colony formation and wound healing assays,we detected the effects of Hsa_circ_0002360 on the activity,proliferation,invasiveness and metastasis Results The expression of Hsa_circ_0002360 was significantly higher in the LAC tissue than in the adjacent tissue(2.84±0.12 vs 1.46±0.08,P<0.01),and so was it in the serum of the LAC patients than in the normal controls(2.59±0.17 vs 1.21±0.11,P<0.01),but remarkably lower in the serum of the patients after than before surgery(1.58±0.09 vs2.65±0.13,P<0.01).Besides,its expression was significantly increased in the LAC A549,H1299 and H1975 cell lines compared with that in the BEAS-2 B cells(P<0.05).In comparison with the si-NC group,the si-circRNA group showed significant decreases in the rates of colony formation([65.66±4.93]%vs[38.33±1.69]%,P<0.01)and migration of the cells([54.66±4.10]%vs[13.50±3.34]%,P<0.01)and in the count of H1975 cells as well(P<0.01).Conclusion Hsa_circ_0002360 can be used as a biomarker for the early diagnosis and treatment of lung adenocarcinoma.