摘要
目的 阐明长链非编码RNA GNAS反义RNA 1(GNAS-AS1)在肺鳞癌(LUSC)进展中的生物学作用和机制。方法 UALCAN在线分析LUSC组织的GNAS-AS1水平,荧光定量PCR检测LUSC细胞的GNAS-AS1水平。向SK-MES-1细胞分别转染靶向GNAS-AS1的小干扰RNA(GNAS-AS1-siRNA)、无关序列siRNA-NC或表皮细胞生长因子EGF(MAPK通路激活剂),分为NC组(阴性对照)、GNAS-AS1-siRNA组(沉默GNAS-AS1表达)和GNAS-AS1-siRNA+EGF组(沉默GNAS-AS1表达+激活MAPK通路)。采用MTT法、划痕实验和Transwell小室实验评估SK-MES-1的增殖、迁移和侵袭能力。Western blotting检测p-MEK2/MEK2和p-ERK1/ERK1水平。结果 GNAS-AS1在LUSC组织中高表达,且与肿瘤分期相关(P<0.05)。与正常肺上皮细胞BEAS-2B相比,LUSC细胞(H1703、H520、H226和SK-MES-1)的GNAS-AS1表达上调(P<0.01)。与NC组相比,GNAS-AS1-siRNA组SK-MES-1细胞的细胞活力、迁移和侵袭能力均降低(P<0.05)。此外,与GNAS-AS1-siRNA组相比,GNAS-AS1-siRNA+EGF组SK-MES-1细胞的细胞活力、迁移和侵袭能力均增强(P<0.05)。GNAS-AS1-siRNA组p-MEK2和p-ERK1水平低于NC组和GNAS-AS1-siRNA+EGF组(P<0.05)。结论 GNAS-AS1通过调节MAPK信号通路活性抑制LUSC进展,提示GNAS-AS1可能是肺癌潜在治疗靶点。
Objective To elucidate the biological role and mechanism of GNAS antisense RNA 1(GNAS-AS1) in lung squamous cell carcinoma(LUSC) progression. Methods UALCAN online database was applied to analyze the GNAS-AS1 level in LUSC tissues, and real-time quantitative PCR was used to detect the GNAS-AS1 levels in LUSC cells. SK-MES-1 cells were transfected with scramble sequence siRNA-NC, small interfering RNA sequence GNAS-AS1-siRNA targeting GNAS-AS1 or activator of MAPK pathway epidermal growth factor(EGF), and allocated into NC group(negative control), GNAS-AS1-siRNA group(Silence of GNAS-AS1) and GNAS-AS1-siRNA+EGF group(Silence of GNAS-AS1 and activation of MAPK pathway). The proliferation, migration, and invasion of SK-MES-1 were detected using MTT assay, scratch assay and Transwell chamber assay, respectively. The expressions of p-MEK2 and p-ERK1 were tested by Western blotting. Results GNAS-AS1 was significantly upregulated in LUSC tissues and related with tumor stage(P<0.05). GNAS-AS1 expression were significantly upregulated in human LUSC cells including H1703, H520, H226 and SK-MES-1 cells compared with normal lung epithelial cells BEAS-2B(P<0.05). The cell viability, migration and invasion of SK-MES-1 cell in GNAS-AS1-siRNA group were weakened compared with NC group(P<0.05). Moreover, the cell viability, migration and invasion of SK-MES-1 cell in GNAS-AS1-siRNA+EGF group were increased compared with GNAS-AS1-siRNA group(P<0.05). The p-MEK2/MEK2 and p-ERK1/ERK1 levels in GNAS-AS1-siRNA group were lower than in NC group and GNAS-AS1-siRNA+EGF group(P<0.05). Conclusion GNAS-AS1 suppressed LUSC progression by regulating ERK/MAPK signaling pathway, indicating that GNAS-AS1 may be a potential therapeutic target in lung cancer.
作者
马虹飞
于青
张莉
尹迪
MA Hongfei;YU Qing;ZHANG Li;YIN Di(Department of Thoracic Surgery,Qingdao Central Hospital,Qingdao 266000,China)
出处
《临床肿瘤学杂志》
CAS
2022年第12期1072-1077,共6页
Chinese Clinical Oncology
关键词
肺鳞癌
GNAS反义RNA
1
侵袭迁移
MAPK信号通路
Lung squamous cell carcinoma
GNAS antisense RNA 1
Invasion and migration
MAPK signaling pathway