H4K16ac Regulates Ras-induced Migration and Proliferation of Breast Cancer Cells

Yaqi Wang^1, , Shunli Zhang¹, Jie Ma¹, Hong Wang¹, Nan Jiang¹, Yang Wang¹, Jun Zhang², Zhuo Wang² and Jinghua Zhang^3,

¹Department of Breast Surgery, Tangshan People's Hospital, Tangshan, China

²Cancer Institute, Tangshan People’s Hospital, Tangshan, China

³Department of Surgical, Tangshan Women and Children's Hospital, Tangshan, China

Cite this paper:
Yaqi Wang, Shunli Zhang, Jie Ma, Hong Wang, Nan Jiang, Yang Wang, Jun Zhang, Zhuo Wang and Jinghua Zhang. H4K16ac Regulates Ras-induced Migration and Proliferation of Breast Cancer Cells. American Journal of Cancer Prevention. 2022; 9(1):10-15. doi: 10.12691/ajcp-9-1-3

Abstract

Ras signal regulates the pathway activity through the extracellular signal regulated kinase (ERK1/2) phosphorylation, Ras mutations or abnormal activation will lead to cancer. In order to study the effects on MCF-7 breast cancer cells proliferation and migration when histone H4K16 acetylating (H4K16ac) in activated Ras signaling pathway, we did a series of experiments. Western blot was used to detect the expression level of ERK1/2, p-ERK1/2, H4 and H4K16ac in different transfected groups. Immunohistochemical technique was used to detect the levels of p-ERK1/2 and H4K16ac in breast cancer tissues and adjacent tissues. The mutant plasmids were constructed to activate cellular Ras signaling pathway and to simulate intracellular H4K16ac. The proliferation ability of cells was detected by CCK-8 technology, and the migration ability of cells was detected by Transwell. In the control group, the activated Ras signal group, and the simulated H4K16ac group, the relative expression levels of p-ERK1/2 were 0.285±0.017, 0.407±0.026, 0.373±0.028; the relative expression levels of H4K16ac were 0.331±0.013, 0.082±0.005, 0.082±0.007. The expression of p-ERK1/2 in breast cancer tissue and adjacent tissue was 0.064±0.001 and 0.051±0.001, respectively; the expression of H4K16ac was 0.028±0.003 and 0.063±0.005, respectively. The proliferation and migration of cancer cells increased by 34.8% and 103.1% respectively (p<0.05), when the Ras signaling pathway was activated. Compared with the cancer cells activated Ras signal pathway, the proliferation and migration of cells simulated H4K16ac were decreased by 25.1% and 42.7% respectively (p<0.05). We demonstrated that after the Ras signal pathway was activated, the expression level of p-ERK1/2 rose, resulting in the decrease of H4K16ac. The increase of H4K16ac will inhibit the Ras signaling pathway activity, thus inhibit proliferation and migration of breast cancer cells. H4K16ac plays an important role in proliferation and migration from breast cancer tissues to normal breast tissues.

Keywords:
breast cancer H4K16ac Ras migration proliferation

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Figures

References:

[1]	XIA C, DONG X, LI H, et al. Cancer statistics in China and United States, 2022: profiles, trends, and determinants [J]. Chinese medical journal, 2022, 135(05): 584-90.
[2]	SONDKA Z, BAMFORD S, COLE C G, et al. The COSMIC Cancer Gene Census: describing genetic dysfunction across all human cancers [J]. Nature Reviews Cancer, 2018, 18(11): 696-705.
[3]	JIN B, LI Y, ROBERTSON K D. DNA methylation: superior or subordinate in the epigenetic hierarchy? [J]. Genes & cancer, 2011, 2(6): 607-17.
[4]	BRUHN C, BASTIANELLO G, FOIANI M. Cancer cell histone density links global histone acetylation, mitochondrial proteome and histone acetylase inhibitor sensitivity [J]. Communications biology, 2022, 5(1): 1-15.
[5]	WEI L, SUN J, ZHANG N, et al. Noncoding RNAs in gastric cancer: implications for drug resistance [J]. Molecular Cancer, 2020, 19(1): 1-17.
[6]	FERNáNDEZ-MEDARDE A, SANTOS E. Ras in cancer and developmental diseases [J]. Genes & cancer, 2011, 2(3): 344-58.
[7]	PRIOR I A, LEWIS P D, MATTOS C. A comprehensive survey of Ras mutations in cancer [J]. Cancer research, 2012, 72(10): 2457-67.
[8]	SCHöNEBORN H, RAUDZUS F, COPPEY M, et al. Perspectives of RAS and RHEB GTPase signaling pathways in regenerating brain neurons [J]. International journal of molecular sciences, 2018, 19(12): 4052.
[9]	SORIANO O, ALCóN-PéREZ M, VICENTE-MANZANARES M, et al. The crossroads between RAS and RHO signaling pathways in cellular transformation, motility and contraction [J]. Genes, 2021, 12(6): 819.
[10]	LIU D, MAO Y, GU X, et al. Unveiling the “invisible” druggable conformations of GDP-bound inactive Ras [J]. Proceedings of the National Academy of Sciences, 2021, 118(11): e2024725118.
[11]	NUSSINOV R, JANG H, GURSOY A, et al. Inhibition of nonfunctional Ras [J]. Cell Chemical Biology, 2021, 28(2): 121-33.
[12]	CARARO-LOPES E, DIAS M H, DA SILVA M S, et al. Autophagy buffers Ras-induced genotoxic stress enabling malignant transformation in keratinocytes primed by human papillomavirus [J]. Cell death & disease, 2021, 12(2): 1-16.
[13]	DESJARLAIS R, TUMMINO P J. Role of histone-modifying enzymes and their complexes in regulation of chromatin biology [J]. Biochemistry, 2016, 55(11): 1584-99.
[14]	CAMPBELL S L, PHILIPS M R. Post-translational modification of RAS proteins [J]. Current Opinion in Structural Biology, 2021, 71(180-92.
[15]	CHENG Y, HE C, WANG M, et al. Targeting epigenetic regulators for cancer therapy: mechanisms and advances in clinical trials [J]. Signal transduction and targeted therapy, 2019, 4(1): 1-39.
[16]	SHVEDUNOVA M, AKHTAR A. Modulation of cellular processes by histone and non-histone protein acetylation [J]. Nature Reviews Molecular Cell Biology, 2022, 23(5): 329-49.
[17]	LEE H-T, OH S, YOO H, et al. The key role of DNA methylation and histone acetylation in epigenetics of atherosclerosis [J]. Journal of lipid and atherosclerosis, 2020, 9(3): 419.
[18]	XIE J, QIAN Y-Y, YANG Y, et al. Isothiocyanate From Moringa oleifera Seeds Inhibits the Growth and Migration of Renal Cancer Cells by Regulating the PTP1B-dependent Src/Ras/Raf/ERK Signaling Pathway [J]. Frontiers in cell and developmental biology, 2021, 9(4): 342-56.
[19]	WANG H, WANG Y, LI D. The PCAF/WSTF/MOF Complex Regulates H3K9ac and H4K16ac in Breast Cancer Cells [J]. American Journal of Cancer, 2022, 9(1): 4-9.
[20]	MARTINELLI E, MORGILLO F, TROIANI T, et al. Cancer resistance to therapies against the EGFR-RAS-RAF pathway: the role of MEK [J]. Cancer treatment reviews, 2017, 53(4): 61-9.
[21]	LIU Y, XING Z B, WANG S Q, et al. MDM 2–MOF–H4K16ac axis contributes to tumorigenesis induced by Notch [J]. The FEBS Journal, 2014, 281(15): 3315-24.
[22]	PUNEKAR S R, VELCHETI V, NEEL B G, et al. The current state of the art and future trends in RAS-targeted cancer therapies [J]. Nature Reviews Clinical Oncology, 2022, 19(10): 637-55.
[23]	MOON A. Ras Signaling in Breast Cancer [M]. Translational Research in Breast Cancer. Springer. 2021: 81-101.
[24]	PENG P-H, LAI J C-Y, CHANG J-S, et al. Induction of epithelial-mesenchymal transition (EMT) by hypoxia-induced lncRNA RP11-367G18. 1 through regulating the histone 4 lysine 16 acetylation (H4K16Ac) mark [J]. American Journal of Cancer Research, 2021, 11(6): 2618.
[25]	ZHOU X, LI T, CHEN Y, et al. Mesenchymal stem cellderived extracellular vesicles promote the in vitro proliferation and migration of breast cancer cells through the activation of the ERK pathway [J]. International journal of oncology, 2019, 54(5): 1843-52.
[26]	ACEVEDO-DíAZ A, MORALES-CABáN B M, ZAYAS-SANTIAGO A, et al. SCAMP3 Regulates EGFR and Promotes Proliferation and Migration of Triple-Negative Breast Cancer Cells through the Modulation of AKT, ERK, and STAT3 Signaling Pathways [J]. Cancers, 2022, 14(11): 2807.