Objective: To investigate the clinical significance and molecular mechanism of chromatin helicase DNA binding protein 5(CHD5) in lung adenocarcinoma(LUAD) and its effect on LUAD cell proliferation, apoptosis and p16INK4a/RB pathway. Methods: UALCAN and Kaplan-Meier Plotter(K-M) databases were used to analyze the expression of CHD5 in LUAD and its relationship with prognosis. Real-time quantitative polymerase chain reaction(qRT-PCR) and Western blotting were used to detect the expression of CHD5 in LUAD tissues, paracancer tissues and cell lines(H1299, H1650, H1975, A549, BEAS-2B). The relationship between CHD5 and clinicopathological features and prognosis of patients were analyzed. The sgRNA vector of CHD5 gene infects H1975 and A549 cells as the overexpression group(CHD5-OE group), and empty vector infected cells as the negative control group(CTRL-OE group), and uninfected cells as the blank group(NC group). The effects of up-regulated of CHD5 expression on the proliferation, apoptosis, migration and invasion of LUAD cells were evaluated by CCK-8, clonogenesis assay, TUNEL apoptosis assay, cell scratching and Transwell assay. Western blotting detected the effect of up-regulation of CHD5 expression on p16INK4a/RB-mediated signaling pathway and cyclin expression. Results: Bioinformatics database analysis showed that CHD5 expression was significantly low in lung adenocarcinoma. Low expression of CHD5 was significantly associated with worse overall survival time(OS). qRT-PCR and Western blotting analysis showed that the expression level of CHD5 in LUAD tissues and cancer cells(H1299, H1650, H1975, A549) was significantly decreased, the difference is statistically significant(P<0.05). Immunohistochemical results showed low or no CHD5 expression in 65.7% lung adenocarcinoma tissue samples. CHD5 expression was related to clinical staging,tumor diameter, lymph node metastasis, but was not related to age, gender, smoking and distant metastasis; Patients with low CHD5 expression had shorter progression-free-survival(PFS). Cox multivariate analysis showed that CHD5 expression was a prognostic factor in LUAD(P<0.001). Overexpression of CHD5 significantly inhibited cell proliferation, migration and invasion in vitro; The expression levels of CyclinD1, CDK4, P-Rb and MMP-9 decreased. The expression levels of Caspase-9, P16 and RB increased. Conclusion: The down-regulated expression of CHD5 in LUAD is related to the poor prognosis of patients. Overexpression of CHD5 can inhibit the proliferation, migration and invasion of LUAD cells, and its mechanism may be related to the inhibition of p16INK4a/RB signaling pathway activation. |
[1] SIEGEL R L,MILLER K D,FUCHS H E, et al. Cancer statistics,2021[J].CA Cancer J Clin,2021,71(1):7-33.
[2] 罗小宁,曾雯,张羽,等.SHCBP1在非小细胞肺癌发生发展中的作用[J].现代医学,2021,49(6):660-665.
[3] 熊磊,刘小龙,徐杨,等.PD-L1在非小细胞肺癌中的表达及其临床意义[J].现代医学,2020,48(9):1169-1173.
[4] WU L,KANG P,TAO S,et al. Genomic profiles and transcriptomic microenvironments in 2 patients with synchronous lung adenocarcinoma and lung squamous cell carcinoma:a case report[J].BMC Med Genomics,2020,13(1):15.
[5] MA Z, SONG J, LIU S, et al. Decreased expression of the CHD5 gene and its clinicopathological significance in breast cancer:correlation with aberrant DNA methylation[J].Oncol Lett,2016,12(5):4021-4026.
[6] WONG R R, CHAN L K, TSANG T P, et al. CHD5 downregulation associated with poor prognosis inepithelial ovarian cancer[J].Gynecol Obstet Invest. 2011,72(3):203-207.
[7] RAMI-PORTA R, BOLEJACK V, CROWLEY J, et al.The iaslc lung cancer staging project:proposals for the revisions of the T descriptors in the forthcoming eighth edition of the tnm classification for lung cancer[J].J Thorac Oncol,2015,10(7):990-1003.
[8] CHANDRASHEKAR D S, BASHEL B, BALASUBRAMANYA S A H, et al. UALCAN:a portal for facilitating tumor subgroup gene expression and survival analyses[J].Neoplasia,2017,19(8):649-658.
[9] LONG S,JI S,XIAO K,et al. Prognostic and immunological value of LTB4R in pan-cancer[J].Math Biosci Eng,2021,18(6):9336-9356.
[10] WU Q,YU L,LIN X,et al. Combination of serum mirnas with serum exosomal mirnas in early diagnosis for non-small-cell lung cancer[J].Cancer Manag Res,2020,12(2):485-495.
[11] PATHAK N,CHITIKELA S,MALIK P S. Recent advances in lung cancer genomics:application in targeted therapy[J].Adv Genet,2021,10(8):201-275.
[12] WU M, JIANG M, XUE M, et al. Epigallocatechin gallate induces CHD5 gene demethylation to promote acute myeloid leukemia cell apoptosis in vitro by regulating p19Arf-p53-p21Cip1 signaling pathway[J].Nan Fang Yi Ke Da Xue Xue Bao,2020,40(9):1230-1238.
[13] HASHIMOTO T, KUROKAWA Y, WADA N, et al. Clinical significance of chromatin remodeling factor CHD5 expression in gastric cancer[J].Oncol Lett,2020,19(1):1066-1073.
[14] HIGASHI M, KOLLA V, IYER R, et al. Retinoic acid-induced CHD5 upregulation and neuronal differentiation of neuroblastoma[J].Mol Cancer,2015, 7(14):150.
[15] ZHU L, WANG R, ZHANG L, et al. RS187960998 polymorphism in miR-211 prevents development of human colon cancer by deregulation of 3'UTR in CHD5[J].Onco Targets Ther,2019,3(12):405-412.
[16] LIU Z, SU D, QI X, et al. MiR500a5p promotes glioblastoma cell proliferation, migration and invasion by targeting chromodomain helicase DNA binding protein 5[J].Mol Med Rep,2018,18(3):2689-2696.
[17] LIU D,LIU X Q,KIEFL R,et al. Effects of the NF-κB pathway agonist IL-1β on non-small cell lung cancer cell lines[J].Ann Clin Lab Sci,2021,51(3):295-301.
[18] GENTILE M,CENTONZA A,LOVERO D,et al. Application of "omics" sciences to the prediction of bone metastases from breast cancer:state of the art[J].J Bone Oncol,2020,2(6):100337.
[19] CHARZEWSKI L, KRZYSKO K A, LESYNG B. Structural characterisation of inhibitory and non-inhibitory MMP-9-TIMP-1 complexes and implications for regulatory mechanisms of MMP-9[J].Sci Rep,2021,11(1):13376.
[20] SUN Y, LUO J, CHEN Y, et al. Combined evaluation of the expression status of CD155 and TIGIT plays an important role in the prognosis of LUAD(lung adenocarcinoma)[J].Int Immunopharmacol,2020,80(1):106198. |