Inhibitory effect of histone H3K27 methylation on proliferation of human well-differentiated nasopharyngeal carcinoma cell line CNE-1

Abstract

Abstract: Objective To investigate the effects of inhibiting histone H3K27me3/2 demethylase on proliferation of well-differentiated human nasopharyngeal carcinoma cell in vitro and explore the underlying mechanisms. Methods Human nasopharyngeal carcinoma cell line CNE-1 was cultured in vitro, and histone H3K27me3/2 demethylation was specifically blocked by histone H3K27me3/2 demethylase inhibitor GSK-J4. The cells were incubated with 0.25,0.5,1.0,2.0,4.0 mmol/L GSK-J4 as the experimental groups, while DMSO was applied as control group. Immunofluorescence assay and flow cytometry were used to analyze the expression of intracellular histone H3K27me3. Real-time quantitative PCR was applied to detect the expression of cyclinD1 and Ki-67 mRNA in cells. Cell counting kit-8 (CCK-8) was used to evaluate cell proliferation. The cloning of CNE-1 cells was observed by colony formation assay. Flow cytometry was also used to detect cell cycle distribution. Results The expression level of histone H3K27me3 in the experimental group was significantly higher than that in the control group (P＜0.05); cyclinD1 and Ki-67 mRNA expression levels in the experimental groups were down-regulated compared with the control group (P＜0.05); The cell viability of CNE-1 cells decreased gradually with the increase of GSK-J4 concentration. The cell proliferation and colony formation in the experimental group were significantly decreased than those in the control group (P＜0.05); cell cycle was blocked in G₀/G₁ phase, the percentage of cells in G₀/G₁ phase was increased and the S phase decreased in the experimental group(P＜0.05). Conclusions Up-regulation of methylation of histone H3K27 in CNE-1 cells induces cell cycle arrest and inhibits cell proliferation. The mechanism may be related to down-regulation of cyclinD1 and Ki-67 mRNA levels.

Key words: histone H3K27, CNE-1, cell proliferation, cell cycle, demethylase inhibitor GSK-J4

CLC Number:

CAI Bo, XING Juan-juan, HU Rong, REN Xin, WANG Rong, WU Tian-ning, GUO Fei, CHEN Shao-qing. Inhibitory effect of histone H3K27 methylation on proliferation of human well-differentiated nasopharyngeal carcinoma cell line CNE-1[J]. Basic & Clinical Medicine, 2020, 40(12): 1613-1618.

References

[1] Chua MLK, Wee JTS, Hui EP, et al. Nasopharyngeal carcinoma[J]. Lancet, 2016, 387: 1012-1024.
[2] Cheng JZ, Chen JJ, Wang ZG, et al. microRNA-185 inhibits cell proliferation while promoting apoptosis and autophagy through negative regulation of TGF-beta1/mTOR axis and HOXC6 in nasopharyngeal carcinoma[J]. Cancer Biomark, 2018, 23: 107-123.
[3] Arcipowski KM, Martinez CA, Ntziachristos P. Histone demethylases in physiology and cancer: a tale of two enzymes, JMJD3 and UTX[J]. Curr Opin Genet Dev, 2016, 36: 59-67.
[4] Barski A, Cuddapah S, Cui K, et al. High-resolution profiling of histone methylations in the human genome[J]. Cell, 2007, 129: 823-837.
[5] Fraineau S, Palii CG, McNeill B, et al. Epigenetic activation of pro-angiogenic signaling pathways in human endothelial progenitors increases vasculogenesis[J]. Stem Cell Reports, 2017, 9: 1573-1587.
[6] Mandal C, Kim SH, Kang SC, et al. GSK-J4-mediated transcriptomic alterations in differentiating embryoid bodies[J]. Mol Cells, 2017, 40: 737-751.
[7] Yan N, Xu L, Wu X, et al. GSKJ4, an H3K27me3 demethylase inhibitor, effectively suppresses the breast cancer stem cells[J]. Exp Cell Res, 2017, 359: 405-414.
[8] 金铁峰, 张美花, 林贞花. 肿瘤表观遗传学[J]. 基础医学与临床, 2011, 31: 204-206.
[9] Cai MY, Tong ZT, Zhu W, et al. H3K27me3 protein is a promising predictive biomarker of patients' survival and chemoradioresistance in human nasopharyngeal carcinoma[J]. Mol Med, 2011, 17: 1137-1145.
[10] Rath BH, Waung I, Camplausen K, et al. Inhibition of the histone H3K27 demethylase UTX enhances tumor cell radiosensitivity[J]. Mol Cancer Ther, 2018, 17: 1070-1078.
[11] Illiano M, Conte M, Sapio L, et al. Forskolin sensitizes human acute myeloid leukemia cells to H3K27me2/3 demethylases GSKJ4 inhibitor via protein kinase A[J]. Front Pharmacol, 2018, 9: 792.
[12] Xie M, Zhao F, Zou X, et al. The association between CCND1 G870A polymorphism and colorectal cancer risk: a meta-analysis[J]. Medicine, 2017, 96: e8269.doi:10.1097/MD.0000000000008269.
[13] Hofstetter C, Kampka JM, Huppertz S, et al. Inhibition of KDM6 activity during murine ESC differentiation induces DNA damage[J]. J Cell Sci, 2016, 129: 788-803.
[14] He QY, Jin F, Li YY, et al. Prognostic significance of downregulated BMAL1 and upregulated Ki-67 proteins in nasopharyngeal carcinoma[J]. Chronobiol Int, 2018, 35: 348-357.
[15] Lee AW, Lin JC, Ng WT. Current management of nasopharyngeal cancer[J].Semin Radiat Oncol, 2012,22:233-244.