ZNF213 facilitates ER alpha signaling in breast cancer cells

doi:10.21203/rs.2.22924/v1

Download PDF

Primary research

ZNF213 facilitates ER alpha signaling in breast cancer cells

https://doi.org/10.21203/rs.2.22924/v1

This work is licensed under a CC BY 4.0 License

You are reading this latest preprint version

Background Breast cancer is the most common women malignancy worldwide, while estrogen receptor alpha positive type accounts for two third of all breast cancers. Although ER alpha positive breast cancer could be effectively controlled by endocrine therapy, more than half of the cases could develop endocrine resistance, making it an important clinical issue in breast cancer treatment. Thus, decoding the detailed mechanism, which controls ER alpha signaling activation and ER alpha protein stability, is of great importance for the improvement of breast cancer therapy.

Methods ZNF213 and ER alpha protein expression level were measured by western blot, while ER alpha target genes were determined by QPCR. WST-1 assay was used to measure cell proliferation. RNA sequence was performed by Ingenuity pathway analysis. The ER alpha signaling activities were measured with luciferase assay, QPCR and western blotting. Protein stability assay and ubiquitin assay were used to determine ER alpha protein degradation and ubiquitination. The immuno-precipation was utilized to determine ER alpha and ZNF213 interaction. The ubiquitin-based immuno-precipitation assay was sued to detect specific ubiquitination manner on ER alpha. The prognostic data of ZNF213 was derived from public available database.

Results Here, we identified ZNF213 as a novel zinc finger protein, which modulates ER alpha protein. ZNF213 expression correlates with poor outcome in endocrine treated patients. ZNF213 depletion inhibits ER alpha signaling and proliferation in breast cancer cells. Further mechanistic studies show that ZNF213 is located in cytosol and nuclear, which modulates ER alpha stability via inhibiting ER alpha K48-linked ubiquitination.

Conclusions Our study reveals an interesting post-translational mechanism between ER alpha and ZNF213 in breast cancer. Targeting ZNF213 could be an appealing strategy for ER alpha positive breast cancer.

Oncology

ZNF213

ER alpha

Breast cancer

Ubiquitin

Stability

Breast cancer ranks NO.1 in women cancer incidence in the world [1]. According to recent cancer statistics, approximately 1.6 million newly diagnosed breast cancer cases each year, which accounts for about 20% of all women cancer incidence. According to the clinical-pathological classification, breast cancer can be divided into three groups: endocrine receptor-positive (estrogen receptor or progesterone receptor), HER2-positive (human epidermal growth factor receptor 2) and triple negative breast cancer (ER alpha, PR and HER2) [2]. Compared with HER2 positive and triple negative breast cancer subtypes, ER alpha positive breast cancer patients show a significant priority in prognosis and could benefit from endocrine therapy [3]. However, more than half of the patients develop endocrine resistance during the treatment, which becomes a major challenge in both the basic research and clinics [4]. Thus, decoding the potential mechanism, which controls ER alpha expression coupled with ER alpha protein stability, is of great importance to characterize endocrine resistance mechanism.

Estrogen receptor alpha (ER) was firstly discovered in 1985, which belongs to the superfamily of nuclear receptors [5]. ER alpha protein is composed of three functional domains: Activation Function 1 (AF1) domain, DNA-binding domain (DBD) and Activation function 2 (AF2) domain [6]. The AF1 domain is responsible for the interaction with co-activators, while the AF2 domain mediates the association with ER alpha ligands. When ER alpha binds to its ligands, the ER alpha protein could trans-locate into the nuclear and form dimers, which subsequently binds to the promoter regions of ER alpha target genes [7]. The aberrant activation of ER alpha signaling has been regarded as the driver pathway for most of ER alpha positive breast cancers [8]. Besides, the activation through ER alpha ligands, the ER alpha signaling could also been modified through post-translational modifications. For example, P300 could induce the acetylation of ER alpha protein at the hinge domain and enhance ER alpha signaling function [9]. Besides, SRC kinase could promote ER alpha phosphorylation at Y537 sites, which induce the conformation change of ER alpha and tamoxifen resistance [10]. Recent studies demonstrated several RING finger proteins could also modulate ER alpha function and breast cancer proliferation [11, 12]. One example is that SHARPIN/RNF31 modulates ER alpha protein stability and ER alpha signaling activity in breast cancer cells [13, 14]. Based on the previous studies, we can propose that the ER alpha interaction proteins might control the ER alpha ubiquitination, protein stability and also ER alpha signaling activity.

Zinc finger domains (ZNF) are small protein motifs, containing about fifty residues [15]. The zinc finger domains modulate the protein-protein interactions [16]. Although ZNF proteins were shown to directly bind DNA/RNA and modulated gene expression [17], several zinc finger proteins were shown to mediate the ubiquitination process and modulate protein stability [12, 14]. Our current study identifies ZNF213 (Zinc finger protein 213) as an important modulator for ER alpha signaling. ZNF213 is elevated in breast cancers and relates to poor prognosis in endocrine treated patients. Besides, ZNF213 facilitates ER alpha signaling and breast cancer progression via enhancing ER alpha stability. Our finding provides a novel insight of ZNF family members in mediation cancer progression and nuclear receptor function. Targeting ZNF213 could be a promising strategy for breast cancer therapy.

Cell culture

MCF-7, T47D and HEK293 cells are got form American Type Culture Collection (ATCC). T47D cells are cultured with RPMI-1640 (42401, Life Technologies) supplemented with 2 mM L-glutamine (25030, Life Technologies) and 10% FBS. MCF-7 and HEK293 are culture with Dulbecco’s Modified Eagle’s Medium that contains 4,5 g/L glucose and 4 mM L-glutamine (DMEM, 41965, Life Technologies) supplemented with 10% Fetal Bovine Serum (FBS, 10270, Life Technologies). All cell lines are characterized by cell line authentication. The cell line authentication via Short Tandem Repeat (STR) is performed via PowerPlex 21 system. The STR data of MCF-7 and T47D cell lines are found consistent with STR data in ATCC.

Plasmids and siRNA

The Myc-ZNF231 plasmid is acquired from Origene Company (https://www.origene.com). The ZNF213 deletion constructs were sub-cloned from the full-length plasmid. The ER alpha full and deletion constructs were described in previous study. The HA-K48 and HA-Ub plasmids were used in previous study. The Estrogen-Response-Element (ERE)-TK reporter and renilla plasmids were used in previous study and are transfected with Lipofectamin 200 (1662298, Invitrogen). For siRNA transfection, the ZNF213 siRNA sequences are #1: 5- GGA UCU CUU CUG GGA CAU AdT dT-3’, 5- UAU GUC CCA GAA GAG AUC CdT dT-3; #2: 5- GGC AUU GGG AGA CAU CCC AdT dT-3, 5- UGG GAU GUC UCC CAA UGC CdT dT-3. The siControl sequences are 5-UUC UCC GAA CGU GUC ACG UTT-3, 5-ACG UGA CAC GUU CG GAGA ATT-3.

RNA extraction and qPCR analysis

RNeasy plus mini kits were used to extract total RNA (Qiagen) [18]. The RNA concentration was measured via Nanodrop. The RNA quality was pre-checked via 18S/28S ratio in 1% agarose gel. Real-time PCR was performed as previously described [19]. 36B4 was used as internal control. Primer sequences for qPCR are provided: GREB1 F: CGT GTG GTG ACT GGA GTA GC, R: ACC TCT TCA AAG CGT GTC GT; ER F: GCT ACG AAG TGG GAA TGA TGA AAG, R: TCT GGC GCT TGT GTT TCA AC; 36B4 F: GGC GAC CTG GAA GTC CA ACT, R: CCA TCA GCA CCA CAG CCT TC; PS2 (TFF1) F: TGG GCT TCA TGA GCT CCT TC, R:TTC ATA GTG AGA GAT GGC CGG.

Quantification of cell viability

MCF-7 and T47D cells were transfected with siZNF213 or siControl in 24-well plate. Twenty-Four hours after transfection, the cells number was countered and 4000 cells were seeded into 96-well plates. The relative cell viability was measured at indicated time points. Cell numbers were determined using the WST-1 cell proliferation reagent as previously described [20].

Western blotting

Cells were harvested and lysed with RIPA buffer. Proteins were separated by electrophoresis on SDS-polyacrylamide gel electrophoresis (PAGE) and electro-transferred to PVDF membrane. The antibodies used in this study were listed here: Anti-ZNF213 (HAP035000, Sigma); Anti-ER alpha (D8H8, 8644, Cell signaling Technology); Anti-HA (MMS-101R, COVANCE); Anti-myc (9E10, ab32, Abcam); Anti-myc (Ab9106, Abcam); Anti-Actin (A5441, Sigma); Anti-Flag (20543-1-AP, Proteintech); Anti-GFP (Ab290, Abcam). Membranes were then washed with PBS for three times and incubated with secondary antibodies Peroxidase-Conjugated AffiniPure Goat Anti-Mouse IgG or Goat Anti-Rabbit IgG. Fluorescent signals were visualized with ECL system. (amersham imager 600, USA).

Luciferase assay

The luciferase activity of estrogen signaling activity was performed using the Dual-Luciferase Reporter kit (Promega, Germany). The ERE luciferase reporter was transfected together with the Renilla plasmid into the cells. Luciferase activity was measured after 24 h.

Co-immunoprecipitation assay

Immunoprecipitation was performed as described in previous study [21]. The MCF-7 total cell lysls were pre-cleared with rabbit IgG for 2 h and subsequently immunoprecipitated with ER alpha antibody (SC8005, Santa Cruz) over night, while rabbit IgG (Santa Cruz) was used as the negative control. The bounded protein was analyzed by Anti-ZNF213 (HAP035000, Sigma). For the overexpression experiment, HEK293 cells were transfected with 5ug GFP-ZNF213 (Full length or deletion domains) and ER alpha plasmid (Full length or deletion domains) in 10 cm dish. Cell lysates were pre-cleared with IgG and subsequently incubate with GFP (Ab290, Abcam) antibody, while rabbit IgG was used as the negative control. The bound proteins were analyzed by western blotting.

Poly-ubiquitination detection assay

To directly detect the enriched overall ubiquitinated or K48-ubiqutinated ER alpha from the cell extracts, HEK293 cells were transfected with 4 ug Ub or 4 ug K48 Ubi plasmid, 2 ug ER alpha together with 0.5 ug Myc-ZNF213 or Myc-vector. After 48 h, total protein was extracted and pre-cleared with 20ul protein A (santa cruz, SC-2001) for 2 h. The supernatant was collected and immunoprecipitated by ER alpha antibody. Western blot with HA antibody was performed to detect total and K48 poly-ubiquitinated ER alpha.

Immunofluorescence assay

MCF-7 cells were fixed with 4% paraformaldehyde in PBS for 10 min, permeabilized with 0.2% Triton X-100 for 5 min, and blocked by 5% BSA in PBS for 1 h. A rabbit anti-ZNF213 (HAP035000, Sigma) and mouse anti-ERα monoclonal antibodies (SC-56833) were used, followed by Alexa Flour 647 (Invitrogen) anti-rabbit antibody and FITC-conjugated anti-mouse antibodies (Jackson ImmunoResearch, West Grove, PA). As negative controls, the samples were incubated with the secondary antibodies without primary antibodies. Images were acquired under conditions fulfilling the Nyquist criterion using Nikon A+ laser scanning confocal system with a 60X oil NA1.4 objective and pinhole size of 1.0 Airy Unit. The acquired pictures were further processed and assembled using ImageJ.

RNA sequence analysis

The global gene expression analysis (siControl and siZNF213) was based on RNA sequencing platform from BGI (Beijing Genomic Institute). The RNA sequence data are deposited in the Gene Expression Omnibus (GEO) database (Assessing number: GSE143948). Analysis was performed for differentially expressed genes (P < 0.01 and fold change > 2) by Ingenuity Pathway Analysis (IPA).

Statistics

Student's t-test, Pearson correlation coefficient, and Cox regression analysis were used for comparisons. A P-value of < 0.05 was considered to be significant.

ZNF213 is elevated in human breast cancer and relates to poor prognosis in endocrine therapy patients

Firstly, we investigate the ZNF213 expression in public available database. From the ONCOMINE database (https://www.oncomine.org), ZNF213 is higher expressed in breast cancer samples compared with normal breast tissues (Fig. 1A-C). Then we analyze the prognostic impact of ZNF213 in breast cancer samples (https://kmplot.com). However, ZNF213 expression does not correlate with the progression-free survival in both ER alpha positive and ER alpha negative patients (Fig. 1D-1E). However, ZNF213 expression specifically relates to poor prognosis in endocrine therapy patients (Fig. 1F).

ZNF213 is required for cell proliferation and ER alpha signaling in breast cancer cells

We further investigate the role of ZNF213 in cell proliferation. The WST-1 assay shows that ZNF213 depletion significantly inhibits cell proliferation in both MCF-7 and T47D cells (Fig. 2A-B). To analyze the role of ZNF213 in breast cancer cells in an unbiased way, we deplete ZNF213 in MCF-7 cells for RNA sequencing analysis. The whole genomic profiling indicates that ZNF213 decreases a group of ER alpha target gene expression (Fig. 2C). This data might indicate ZNF213 modulates ER alpha signaling in breast cancer cells. In order to prove this hypothesis, we use two independent siRNAs to carry out the experiments. Figure 2D shows that the siRNAs show high knockdown efficiency in MCF-7 cells. The immuno-blotting shows that ZNF213 depletion significantly decreases ER alpha protein level (Fig. 2E). Besides, QPCR assay shows that ZNF213 depletion decreases the expression of ER alpha target genes, including GREB1 and PS2 (Fig. 2F).

ZNF213 depletion inhibits ER alpha protein and ER alpha target genes in breast cancer cells

We further test ZNF213 effect on ER alpha signaling in both vehicle and E2-treated conditions. ZNF213 depletion could decrease ER alpha protein level in vehicle and E2-treated conditions in both MCF-7 and T47D cells (Fig. 3A-B). In order to determine if ZNF213 knockdown could affect ER alpha transcriptional activity, we measure estrogen response element (ERE) luciferase activity in both MCF-7 and T47D cells. The luciferase assay shows that ZNF213 depletion decreases ERE luciferase activity in both MCF-7 and T47D cells (Fig. 3C-D). Consistently, ZNF213 depletion could dramatically decrease ER alpha target gene expression in MCF-7 cells, including PS2, GREB1 and PDZK1 (Fig. 3E).

ZNF213 could associate with ER alpha in breast cancer cells

We further investigate the localization of ZNF213 and ER alpha in breast cancer cells. The immuno-staining shows that ER alpha is mainly located in the nuclear, while ZNF213 locates both in the cytosol and nuclear (Fig. 4A). The endogenous immuno-precipitation shows that ZNF213 could interact with ER alpha in MCF-7 cells (Fig.4B). ER alpha is composed of three functional domains: AF1 domain, DNA binding domain and AF2 domain (Fig. 4C), while ZNF213 is composed by LeR/SCAN domain, KRAB-A domain and ZF domain (Fig. 4D). We make the deletion constructs of ER alpha and ZNF213 to further characterize the interaction domain. The immuno-precipitation assay shows that the AF1 domain of ER alpha is required for its interaction with ZNF213, while the ZF domain of ZNF213 is responsible to associate with ER alpha (Fig. 4E-G).

ZNF213 modulates ER alpha poly-ubiquitination and protein stability

Since ZNF213 could associate with ER alpha in breast cancer cells, we further investigate the biological effect of such interaction. Co-transfection of ER alpha and ZNF213 in HEK293 cells shows that ZNF213 could increase ER alpha protein level, which effect could be minimized with the presence of the proteasome inhibitor MG132 (Fig. 5A). The protein half-life assay shows that ZNF213 could increase the protein stability of ER alpha (Fig. 5B-5C), while co-transfection with ZNF213 deletion variants shows that the ZF domain is required for the stabilization effect for ER alpha protein (Fig. 5D). We further investigate the effect of ZNF213 on ER alpha ubiquitination. The ubiquitination assay shows that ZNF213 could significantly inhibit ER alpha overall ubiquitination level and K48-linked ubiquitination (Fig. 5E-F).

In this study, we identify one znic finger protein ZNF213, which is higher expression in human breast cancer samples, promotes ER alpha signaling activity and ER alpha stability in breast cancer cells. ZNF213 associates with ER alpha and inhibits ER alpha poly-ubiquitination and degradation (Fig. 6).

The importance of ER alpha signaling has been identified for 30 years [22], based on the fact that ER alpha is one of the most important drivers for breast cancer progression. Although ER alpha mutation is not common, the elevated expression of ER alpha could be found in most of human breast cancers [8]. Based on current knowledge, ER alpha signaling is a suitable target for breast cancer therapy. Selective ER alpha modulators, such as tamoxifen, are effective in blocking ER alpha signaling and breast cancer progression. However, most of patients will development endocrine resistance, making it a critical clinical issue for breast cancer therapy. Based on the published literatures, most of resistant breast tumors still maintain ER alpha expression [23]. There are several possible mechanisms for endocrine resistance. For example, ER alpha could trans-activate other growth factor pathways, such as IGF and EGFR signaling, to overcome endocrine resistance [24]. Besides, several modifications, including ubiquitin, acetylation and phosphorylation, could enhance ER alpha signaling strength and overcome tamoxifen resistance [13, 25, 26]. Base on these mechanisms, modulating ER alpha protein stability could a plausible strategy for therapeutics in ER alpha positive breast cancer patients.

Recently studies have identified several RING finger proteins or Znic finger proteins in modulating ER alpha stability and breast cancer progression. For example, RNF31 could associate with ER alpha and promote ER alpha stability via inducing ER alpha mono-ubiquitination [27]. Besides, RNF8 was also found to co-activate ER alpha target genes and prolong ER alpha half-life in breast cancer cells [12]. When it comes to ZNF213, which belongs to zinc finger protein members, we firstly identified its role in breast cancer. ZNF213 was firstly regarded as a modulator for gene expression, based on its capability to associate with DNA [28]. However, our study showed that ZNF213 could associate with ER alpha protein and promotes ER alpha function. The co-transfection in HEK293 cells indicate the modulation effect of ZNF213 on ER alpha signaling goes through protein stability, not through direct genomic regulation. These findings provide a novel insight of ZNF family members in modulating estrogen signaling and breast cancer progression. ZNF231 could be a novel target for ER alpha positive breast cancer patients.

In summary, our study provides a novel regulatory mechanism between ZNF213 and ER alpha in breast cancer cells. The zinc finger protein ZNF213 modulates ER alpha signaling and breast cancer progression through a post-translational mechanism. Our study implicates the important role of ZNF213 in ER alpha signaling and improves the understanding of ZNF213 in both genomic and non-genomic regulation in human cancer. As such an important regulator of ER alpha signaling, ZNF213 could be a promising target for ER alpha positive breast cancer therapeutics.

ZNF213: Zinc finger protein 213

AF1: Transcriptional activation domain 1

DBD: DNA binding domain

AF2: Transcriptional activation domain 2

ER alpha: Estrogen receptor alpha

HER2: Human epidermal growth factor receptor 2

PR: Progesterone receptor

TBNC: Triple negative breast cancer

RBR: RING-between-RING domain

ZF: Zinc finger domain

EMT: Epithelial-mesenchymal transition

ATCC: American Type Culture Collection

Bray F et al: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: a cancer journal for clinicians 2018, 68(6):394-424.
Onitilo AA, Engel JM, Greenlee RT, Mukesh BN: Breast cancer subtypes based on ER/PR and Her2 expression: comparison of clinicopathologic features and survival. Clinical medicine & research 2009, 7(1-2):4-13.
Vaz-Luis I et al: Outcomes by tumor subtype and treatment pattern in women with small, node-negative breast cancer: a multi-institutional study. Journal of clinical oncology : official journal of the American Society of Clinical Oncology 2014, 32(20):2142-2150.
Massarweh S, Schiff R: Resistance to endocrine therapy in breast cancer: exploiting estrogen receptor/growth factor signaling crosstalk. Endocrine-related cancer 2006, 13 Suppl 1:S15-24.
Walter P et al: Cloning of the human estrogen receptor cDNA. Proceedings of the National Academy of Sciences of the United States of America 1985, 82(23):7889-7893.
Kumar R et al: The dynamic structure of the estrogen receptor. Journal of amino acids 2011, 2011:812540.
Paige LA et al: Estrogen receptor (ER) modulators each induce distinct conformational changes in ER alpha and ER beta. Proceedings of the National Academy of Sciences of the United States of America 1999, 96(7):3999-4004.
Shaaban AM, Sloane JP, West CR, Foster CS: Breast cancer risk in usual ductal hyperplasia is defined by estrogen receptor-alpha and Ki-67 expression. The American journal of pathology 2002, 160(2):597-604.
Wang C et al: Direct acetylation of the estrogen receptor alpha hinge region by p300 regulates transactivation and hormone sensitivity. The Journal of biological chemistry 2001, 276(21):18375-18383.
Osborne CK et al: Role of the estrogen receptor coactivator AIB1 (SRC-3) and HER-2/neu in tamoxifen resistance in breast cancer. Journal of the National Cancer Institute 2003, 95(5):353-361.
Liu Z et al: RNF168 facilitates oestrogen receptor a transcription and drives breast cancer proliferation. Journal of cellular and molecular medicine 2018.
Wang S et al: RNF8 identified as a co-activator of estrogen receptor alpha promotes cell growth in breast cancer. Biochimica et biophysica acta Molecular basis of disease 2017, 1863(6):1615-1628.
Zhuang T et al: SHARPIN stabilizes estrogen receptor alpha and promotes breast cancer cell proliferation. Oncotarget 2017, 8(44):77137-77151.
Zhu J et al: The atypical ubiquitin ligase RNF31 stabilizes estrogen receptor alpha and modulates estrogen-stimulated breast cancer cell proliferation. Oncogene 2014, 33(34):4340-4351.
Chen X et al: Identification and characterization of a zinc finger gene (ZNF213) from 16p13.3. Biochimica et biophysica acta 1999, 1444(2):218-230.
Laity JH, Lee BM, Wright PE: Zinc finger proteins: new insights into structural and functional diversity. Current opinion in structural biology 2001, 11(1):39-46.
Bomar MG, Pai MT, Tzeng SR, Li SS, Zhou P: Structure of the ubiquitin-binding zinc finger domain of human DNA Y-polymerase eta. EMBO reports 2007, 8(3):247-251.
Xue M et al: Regulation of estrogen signaling and breast cancer proliferation by an ubiquitin ligase TRIM56. Oncogenesis 2019, 8(5):30.
Yang H et al: SMURF1 facilitates estrogen receptor a signaling in breast cancer cells. Journal of experimental & clinical cancer research : CR 2018, 37(1):24.
Zhu J et al: RING finger protein 31 promotes p53 degradation in breast cancer cells. Oncogene 2016, 35(15):1955-1964.
Yang H et al: SHARPIN Facilitates p53 Degradation in Breast Cancer Cells. Neoplasia 2017, 19(2):84-92.
Greene GL et al: Sequence and expression of human estrogen receptor complementary DNA. Science 1986, 231(4742):1150-1154.
Mills JN, Rutkovsky AC, Giordano A: Mechanisms of resistance in estrogen receptor positive breast cancer: overcoming resistance to tamoxifen/aromatase inhibitors. Current opinion in pharmacology 2018, 41:59-65.
Shou J et al: Mechanisms of tamoxifen resistance: increased estrogen receptor-HER2/neu cross-talk in ER/HER2-positive breast cancer. Journal of the National Cancer Institute 2004, 96(12):926-935.
Anbalagan M, Rowan BG: Estrogen receptor alpha phosphorylation and its functional impact in human breast cancer. Molecular and cellular endocrinology 2015, 418 Pt 3:264-272.
Zhuang T et al: p21-activated kinase group II small compound inhibitor GNE-2861 perturbs estrogen receptor alpha signaling and restores tamoxifen-sensitivity in breast cancer cells. Oncotarget 2015, 6(41):43853-43868.
Zhu J et al: Atypical ubiquitin ligase RNF31: the nuclear factor modulator in breast cancer progression. BMC cancer 2016, 16:538.
Eom KS, Cheong JS, Lee SJ: Structural Analyses of Zinc Finger Domains for Specific Interactions with DNA. Journal of microbiology and biotechnology 2016, 26(12):2019-2029.

Declarations: There is nothing to declear.

Ethics Approval and Consent to participate: No ethic issues involves in the study.

Consent for publication: All the authors consent for publication.

Availability of supporting data: The RNA sequence data are deposited in the Gene Expression Omnibus (GEO) database (Assessing number: GSE143948)

Competing Interests: The authors declare no conflict of interest.

Funding: The project was supported from the National Natural Science Foundation of China (No. 81702725, Ting Zhuang); The National Natural Science Foundation of China (Grant No. U1804167, NO.81770721; NO.81570624, Qingsong Huang); Key Scientific and Technological Projects of Henan Province (Grant No. 202102310024, Qingsong Huang).

Authors’ contributions: T.Z. and HJ. Y performed most of the bench work. QS. H, and ZG. N supervised the process of the study and performed the manuscript writing. X.L. and WC. Z participated in western blot, real time PCR work. J.Z performed the prognosis data analysis.

Acknowledgement: We thank all the members of Laboratory of Molecular Oncology in Xinxiang University for sharing valuable material and research support.

Download PDF

Editorial decision: Major revision
30 Mar, 2020
Review #2 received at journal
29 Mar, 2020
Reviewer #2 agreed at journal
21 Mar, 2020
Review #1 received at journal
16 Feb, 2020
Reviewer #1 agreed at journal
10 Feb, 2020
Reviewers invited by journal
09 Feb, 2020
Editor assigned by journal
06 Feb, 2020
Submission checks completed at journal
05 Feb, 2020
Editor invited by journal
05 Feb, 2020
First submitted to journal
04 Feb, 2020

You are reading this latest preprint version

ZNF213 facilitates ER alpha signaling in breast cancer cells

Status:

Version 1

Abstract

Figures

Background

Materials And Methods

Results

Discussion

Conclusions

Abbreviations

References

Declarations

Status:

Version 1