Expression of FGD6 in lung adenocarcinoma and its effect on cell proliferation,migration and invasion
10.3781/j.issn.1000-7431.2024.2310-0593
- VernacularTitle:FGD6在肺腺癌中的表达情况及其对肺腺癌细胞增殖、迁移和侵袭的影响
- Author:
Yingna LIAO
1
;
Liheng QIAN
1
;
Shuxin LI
1
;
Huizhen NIE
1
Author Information
1. 上海交通大学医学院附属仁济医院上海肿瘤研究所肿瘤系统医学全国重点实验室,上海 200240
- Publication Type:Journal Article
- Keywords:
Lung adenocarcinoma;
Cell proliferation;
Cell apoptosis;
Cell migration;
Cell invasion;
Hippo pathway
- From:
Tumor
2024;44(4):319-333
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the expression of FGD6(FYVE,RhoGEF,and PH domain containing 6)in lung adenocarcinoma(LUAD)tissues and explore its impact on LUAD cells proliferation,apoptosis,migration,invasion,and the possible mechanism.Methods:By searching bioinformatics websites and databases,the differences in the expression of FGD6 gene in LUAD,paracancerous tissues and normal lung tissues were analyzed,and further analyzed the correlation between the expression of FGD6 and the survival prognosis of LUAD patients as well as the correlation with the clinicopathological features of LUAD patients.Two different siRNAs(si-FGD6-1,si-FGD6-2)targeting FGD6 gene were transfected into LUAD cell lines(A549 and NCI-H23)using lipofection.The knockdown efficiency of the siRNAs was detected by real-time fluorescence quantitative PCR and Western blotting.CCK-8 assay,clone formation assay and FCM assay were used to detect the effects of silencing FGD6 expression on the proliferation and apoptosis of A549 and NCI-H23 cells;Wound healing assay assay and Transwell assay were used to detect the effects of silencing FGD6 expression on the migratory and invasive ability of A549 and NCI-H23 cells.Subsequently,Gene Set Enrichment Analysis(GESA)was performed based on the transcriptome data of LUAD patients in The Cancer Genome Atlas(TCGA)database,and it was hypothesized that FGD6 might regulate the Hippo signaling pathway in LUAD cells.Real-time fluorescence quantitative PCR and Western blotting were used to detect the effects of silencing FGD6 expression on the connective tissue growth factor(CTGF)and cysteine-rich 61(CYR61)genes and the phosphorylation of the Hippo signaling pathway downstream in LUAD cells.The effect of the expression level of phosphorylated Yes-associated protein(YAP)in the Hippo signaling pathway.Results:Database analysis showed that FGD6 expression was up-regulated in a variety of tumors,and the expression level in LUAD was significantly higher than that in normal tissues(P<0.001).The overall survival time of patients with high expression of FGD6 was less than that of those with low expression(P<0.05),and it was positively correlated with the clinical stage and lymph node metastatic status of LUAD patients.After knocking down FGD6 expression,the expression levels of FGD6 mRNA and protein in A549 and NCI-H23 cells were significantly down-regulated,and their cell proliferation,anti-apoptosis,migration and invasion abilities were significantly reduced(all P<0.01).The results of real-time fluorescence quantitative PCR showed that knockdown of FGD6 expression led to significant down-regulation of the expression levels of CTGF and CYR61 mRNA,the downstream targeting factors of the Hippo signaling pathway(both P<0.01),while the results of Western blotting showed that the expression levels of phosphorylated Yes-associated protein(p-YAP)were significantly down-regulated in the Hippo signaling pathway(P<0.01).Conclusion:FGD6 is highly expressed in LUAD tissues,and its high expression is associated with poor prognosis.Knockdown of FGD6 gene can inhibit LUAD cells proliferation,apoptosis,migration and invasion,and activate the Hippo pathway,suggesting its potential as a therapeutic target in LUAD.
