Breast cancer occurring during pregnancy of Luminal A type overexpressed CXCL13

doi:10.21203/rs.3.rs-3145411/v1

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Breast cancer occurring during pregnancy of Luminal A type overexpressed CXCL13

https://doi.org/10.21203/rs.3.rs-3145411/v1

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Background

Breast cancer during pregnancy and lactation occurs in 2.6% of female patients aged < 45 years, and its incidence has been increasing. The pathogenesis and genomic profiles of pregnancy-associated breast cancer (PABC) remain unclear. We analyzed patients with breast cancer that occurred during pregnancy (PrBC) and compared the findings with those of patients with breast cancer that did not occur during pregnancy (non-PrBC).

Methods

We performed gene expression analyses (using microarray and qRT-PCR) of PrBC (8 and 22 samples, respectively) and non-PrBC (8 and 24 samples, respectively) groups, which were matched for age, hormone receptor expression, and HER2 expression.

Results

Microarray analysis showed that 355 genes were upregulated and 151 genes were downregulated in the luminal type PrBC group compared to those in the non-PrBC group. Chemokine, C-X-C motif chemokine ligand 13 (CXCL13) gene was the most upregulated in the PrBC group compared to that in the non-PrBC group (log2 fold-change = 101.2), especially in the luminal A-type cells (p = 0.016). This result was corroborated by the qRT-PCR analysis of microdissected cancer cells, which was conducted using formalin-fixed paraffin-embedded (FFPE) samples (p < 0.001). A negative correlation was observed between CXCL13 and estrogen receptor 1 (ESR1) mRNA expression levels in luminal A-type breast carcinoma (τ = -0.109; p < 0.001).

Conclusion

CXCL13, a chemokine that regulates the function of immune cells, was significantly overexpressed in PrBC cells. The expression levels of CXCL13 and estrogen receptor (ER) were inversely correlated.

breast cancer

breast cancer during pregnancy (PrBC)

CXCL13

pregnancy

pregnancy-associated breast cancer (PABC)

Breast cancer during pregnancy and lactation occurs in 2.6% of female patients under the age of 45 [1], but the frequency is gradually increasing as women are postponing pregnancy [2]. Gestational breast cancer has long been referred to as pregnancy-associated breast cancer (PABC), which is defined as breast cancer diagnosed during pregnancy, within the first year of childbirth, or during breastfeeding. In recent years, it has become clear that breast cancer that occurs during pregnancy (PrBC) and breast cancer that occurs during the postpartum period (PPBC) have different biological characteristics and prognoses; therefore, it is important to differentiate PrBC from PPBC when discussing PABC [3]. The most common cancers that occur during pregnancy include hematologic malignancies, breast, thyroid, colon, cervical, and ovarian cancers, and melanoma. Breast cancer affects approximately one to three in 10,000 pregnancies [4].

PABC has long been considered to have a poor prognosis. Two clinical studies on PABC conducted in Japan showed a significantly lower overall survival (OS) in patients with PABC than that in the control group [5, 6]. However, recent reports have suggested that the prognosis of PABC is not necessarily poor when the age and progression of patients with PABC are compared with those of patients with non-pregnancy-related breast cancer because of the progress in aggressive therapeutic intervention [7–9]. Kataoka reported that patients with breast cancer diagnosed within one year of childbirth or during lactation (LBC) had significantly lower breast cancer-specific disease-free survival (BC-DFS) and OS than those in patients with PrBC [10]. Few genomic studies on PABC have been conducted [11], and its molecular characteristics remain unknown.

To understand the biological features of PrBC, we categorized patients into two groups comprising those with PrBC and those with breast cancer that did not occur during pregnancy (non-PrBC). We analyzed and compared ages and hormone receptor and HER2 receptor expression levels of the patients in the two groups.

Patients and ethics statement

We retrospectively reviewed the medical records of 45 women with primary PrBC who underwent surgery without neoadjuvant therapy at St. Luke’s International Hospital between January 2005 and December 2017. Ages, estrogen receptor (ER) or progesterone receptor (PGR) levels, and HER2 expression levels of 45 female patients with primary non-PrBC were compared with those of patients with PrBC. ER and PGR were defined as positive when the Allred score was equal to or more than 3. HER2 positivity was evaluated according to the 2018 ASCO/CAP guidelines 2018 [12]. The patient characteristics are presented in Table 1. In this study, luminal A-type breast cancer was defined as ER- or PGR-positive, HER2-negative, and Ki-67 labeling index less than 30%, whereas luminal B-type breast cancer was defined as ER-positive, HER2-negative, and Ki-67 labeling index greater than 30%, according to the recommendations of the St. Gallen Consensus Conference held in 2015 [13]. The study design was approved by the Institutional Review Board of St. Luke’s International Hospital (approval number 18-R085) according to the Declaration of Helsinki, and written informed consent was obtained from all participants.

Table 1

Clinicopathological characteristics of the patients.
	non-PrBC	PrBC
	(n = 45)	(n = 45)	p-value
Age
25–34	10	10	Match
35–44	35	35
Years of surgery
2005–2017	45	45	Match
Primary tumor (T)
T1	27	21	0.37
T2	14	21
T3	3	3
T4	1	0
Regional lymph node (N)
N0	31	27	0.78
N1<	12	12
Histological grade
I	13	9	0.58
II	20	21
III	12	15
Tubule formation
Score 1	12	2	0.01
Score 2	8	12
Score 3	25	31
Nuclear atypia
Score 1	1	2	0.84
Score 2	34	35
Score 3	10	8
Mitotic counts
Score 1	18	18	0.7
Score 2	12	9
Score 3	15	18
Nuclear grade
I	20	18	0.67
II	11	9
III	14	18
Estrogen receptor
Positive	39	39	Match
Negative	6	6
HER2
Positive	8	8	Match
Negative	37	37
Subtypes
ER+/HER2- (LA Ki67: <30)	24	22	0.99
ER+/HER2- (LB Ki67: >=30)	8	10
ER+/HER2+	7	7
ER-/HER2+	1	1
ER-/HER2-	5	5
Ki-67 labeling Index
Mean value	29.5	33.4	0.32

RNA microarray analysis

Microarray analysis was performed on eight patients with PrBC (four cases of luminal A type and four cases of luminal B type) and eight patients without PrBC (five cases of luminal A type and three cases of luminal B type). Fresh tissue samples were obtained and stored in RNAlater stabilization solution (AM7022, Thermo Fisher Scientific Inc., MA, USA) at -80 ºC until use. Total RNA extraction and mRNA microarray analysis were performed at Macrogen Japan Corp. (Tokyo, Japan) using the GeneChip Human Gene 2.0 ST Array (#902499, Thermo Fisher Scientific Inc.). The data were normalized using the Robust Multichip Analysis (RMA) algorithm available in Affymetrix Power Tools (http://media.affymetrix.com/support/developer/powertools/changelog/index.html). The genes were classified into two categories based on the fold-change (FC) in expression levels: upregulated genes (log2FC ≥ 1.5) and downregulated genes (log2FC ≤ 1.5).

RNA extraction and quantitative RT-PCR assays

RT-PCR was performed using all luminal A-type PrBC (n = 22) and non-PrBC (n = 24) samples as controls. Formalin-fixed paraffin-embedded (FFPE) tissue blocks were cut (thickness of sections = 8 µm) and mounted on membrane film-coated glass slides. The paraffin sections were deparaffinized, hydrated, and stained with toluidine blue. After air-drying at room temperature, the breast carcinoma cells were microdissected using a laser microdissection (LMD) instrument (LMD 6000, Leica Microsystems, Tokyo, Japan). Total RNA extraction and cDNA synthesis were performed using microdissected samples as previously described [14]. Although RNA was extracted from all 46 FFPE luminal A-type breast cancer samples, one non-PrBC sample was excluded from the analysis because of unsuitable RNA quality. RT-PCR analysis was performed using the StepOnePlus Real-Time PCR System (#4376600, Thermo Fisher Scientific Inc.) using 96-well plates, and 10 µl of the reaction mixture comprised 5 µl TaqPath™ qPCR Master Mix, CG (#A15297, Thermo Fisher Scientific Inc.), 0.5 µl each of the 20X TaqMan probes (CXCL13: Hs00757930_m1, ESR1: Hs01046816_m1, PGR: Hs01556702_m1, and β-actin: Hs01060665_g1, #4453320), 3.5 µl of double-distilled water, and 1 µl of the cDNA template. PCR was performed by preheating the reaction mixtures at 95°C for 60 s, followed by 50 cycles of 95°C for 15 s and 60°C for 60 s. Relative quantification was performed using the ΔΔCt method, and the expression level of CXCL13 mRNA was normalized according to the β-actin expression level.

Statistical analysis

The chi-square test or the Fisher's exact test was used to compare clinicopathological characteristics of the PrBC and the non-PrBC groups. The significance of the mRNA expression levels measured by qRT-PCR was analyzed using the Mann–Whitney U test. The correlation was calculated using Kendall's tau correlation coefficient. Statistical analyses were performed using R version 4.2.2 (www.r-project.org) and BellCurve for Excel (Social Survey Research Information Co., Ltd., Tokyo, Japan). Microarray data were exported for gene-level RMA analysis, and differentially expressed genes (DEGs) were examined. Hierarchical cluster analysis was performed on the basis of Euclidean distance and complete linkage, which clustered similar probe sets and samples according to the expression levels (normalized values) of genes showing significant results. The statistical significance of the signal expression data was determined using the Mann–Whitney U test and fold-change values. The false discovery rate (FDR) was controlled by adjusting the p value using the Benjamini–Hochberg algorithm. Hierarchical cluster analysis was performed using R version 4.2.2.

Clinicopathological characteristics of patients with PrBC and non-PrBC

The clinicopathological characteristics of all examined patients (PrBC, n = 45; non-PrBC, n = 45) are summarized in Table 1. Lymph node dissection was not performed in six patients in the PrBC group and two patients in the non-PrBC group. The PrBC group had a lower tubule formation score than that of the non-PrBC group (p = 0.01).

RNA microarray analysis of PrBC and non-PrBC cells of luminal type

In the PrBC group, 355 genes were upregulated (log2FC 1.5) and 151 genes were downregulated (log2FC -1.5) compared to that in the non-PrBC group of patients with the luminal type of cancer (Supplementary Table S1). CXCL13 was the most upregulated gene in the PrBC group (log2FC 6.5). CXCL13 was significantly upregulated in the luminal A-type PrBC group (log2FC 101.2; p = 0.016) but was downregulated in the luminal B type, although there were no significant differences in the expression levels (log2FC -4.2; p = 0.400). ESR1 and PGR genes were downregulated in the PrBC group (log2FC -2.0 and − 3.1, respectively), especially in the luminal A-type PrBC group, and showed statistically significant differences in expression levels (log2FC -2.9 and − 5.2 and p = 0.032 and 0.016, respectively). Overall, different trends were observed in the luminal A and luminal B groups. In this study, we focused on the luminal A group.

Clinicopathological characteristics of patients with luminal A-type PrBC and non-PrBC

The clinicopathological characteristics of the luminal A-type cancers (22 cases of PrBC and 24 cases of non-PrBC) are summarized in Table 2. Lymph node dissection was not performed in one patient in the PrBC group and two patients in the non-PrBC group. The PrBC group had a lower tubule formation score than that of the non-PrBC group (p = 0.03). Although the non-PrBC group was matched for overall ER positivity/negativity, the Allred scores of ER expression in the two luminal A groups were significantly different (p = 0.01), indicating a low expression level of ER in the PrBC group.

Table 2

Clinicopathological characteristics of patients with luminal A-type breast cancer.
	non-PrBC	PrBC
	(n = 24)	(n = 22)	p-value
Age
25–34	4	3	1.00
35–44	20	19
Years of surgery
2005–2017	24	22	Match
Primary tumor (T)
T1	17	12	0.24
T2	4	9
T3	2	1
T4	1	0
Regional lymph node (N)
N0	16	11	0.58
N1<	7	9
Histological grade
I	12	7	0.30
II	12	14
III	0	1
Tubule formation
Score 1	10	2	0.03
Score 2	6	6
Score 3	8	14
Nuclear atypia
Score 1	1	2	0.30
Score 2	23	18
Score 3	0	2
Mitotic counts
Score 1	16	15	1.00
Score 2	5	5
Score 3	3	2
Nuclear grade
I	18	15	0.25
II	6	4
III	0	3
Estrogen receptor
Allred score
TS0	0	1	0.01
TS4	0	3
TS6	3	6
TS7	5	7
TS8	16	5
Progesterone receptor
Allred score
TS5	2	1	0.59
TS6	0	1
TS7	6	8
TS8	16	12
Ki-67 labeling Index
Mean value	13.5	16.4	0.25

Quantitative analyses of CXCL13, ESR1, andPGRmRNA expression levels in luminal A-type PrBC and non-PrBC cells

High expression levels of CXCL13 mRNAs and low expression levels of ESR1 and PGR mRNAs were observed in the luminal A-type PrBC group using the microarray method. These findings were validated using qRT-PCR. The CXCL13 mRNA expression level was significantly higher in the PrBC group than that in the non-PrBC group (Fig. 1a; p < 0.001). ESR1 and PGR mRNA expression levels were significantly lower in the PrBC group than those in the non-PrBC group (Fig. 1b and 1c; p < 0.001 and = 0.0013, respectively).

Relation between CXCL13 and ESR1 mRNA expression levels determined by microarray and qRT-PCR analyses of luminal A-type PrBC and non-PrBC samples

We examined the correlation between CXCL13 and ESR1 mRNA expression levels in luminal A-type breast carcinoma using a microarrays and qRT-PCR. A strong, significant inverse correlation (τ = -0.852, p = 0.037) was observed between CXCL13 and ESR1 mRNA signal intensities on microarray analysis of luminal A-type breast carcinomas (Fig. 2a; Table 3). A weak, significant inverse correlation (τ = -0.109, p < 0.001) was observed between CXCL13 and ESR1 mRNA signal intensities on qRT-PCR analysis of luminal A-type breast carcinomas (Fig. 2b; Table 3).

Table 3

Correlation between *CXCL13* and *ESR1* or *PGR* mRNA expression levels.
		CXCL13 (microarray)			CXCL13 (qRT-PCR)
Target (method)	n	τ (correlation coefficient)	Strength of Correlation	p value	τ (correlation coefficient)	Strength of Correlation	p value
ESR1 (microarray)	9	-0.852	Strong	0.037	-	-	-
ESR1 (RT-PCR)	45	-	-	-	-0.109	Weak	< 0.001
PGR (microarray)	9	-0.611	Moderate	0.211	-	-	-
PGR (RT-PCR)	45	-	-	-	-0.154	Weak	0.183

Relation between CXCL13 and PGR mRNA expression levels determined by microarray and qRT-PCR analyses of luminal A-type PrBC and non-PrBC samples

We examined the correlation between CXCL13 and PGR mRNA expression levels in luminal A-type breast carcinoma using a microarrays and qRT-PCR. A moderate, but no significant inverse correlation (τ = -0.611, p = 0.211) was observed between CXCL13 and PGR mRNA signal intensities on microarray analysis of luminal A-type breast carcinomas (Fig. 2c; Table 3). A weak, but no significant inverse correlation (τ = -0.154, p = 0.183) was observed between CXCL13 and PGR mRNA signal intensities on qRT-PCR analysis of luminal A-type breast carcinomas (Fig. 2d; Table 3).

Hierarchical cluster analysis of luminal A-type PrBC and non-PrBC samples

Hierarchical cluster analysis showed that PrBC and non-PrBC groups were separated into different clusters (Fig. 3). CXCL13 belongs to a gene cluster that is overexpressed in PrBCs and underexpressed in non-PrBCs. ESR1 and PGR belong to a gene cluster that is underexpressed in PrBCs and overexpressed in non-PrBCs (Fig. 3). The genes comprising these clusters are shown in Supplementary Fig. S1.

In recent years, the clinical characteristics of PABC have become clear. Several studies have shown that PABC is associated with a significantly higher risk of breast cancer death than that associated with non-PABC [15, 16]. Moreover, PPBC has a poorer prognosis compared to that of PABC [15, 17]. A recent Japanese cohort study has shown that patients with PPBC have significantly lower BC-DFS and OS rates than those in patients with PrBC [10]. However, the pathogenesis and genomic profiles of PABC remain unclear [11]. Our study revealed that CXCL13 was highly expressed in PrBCs, especially in luminal A-type breast cancer, and that high expression of CXCL13 was inversely correlated with low expression of ER and PGR.

CXCL13 is a B-cell activating chemokine produced by lymph node stromal cells and follicular dendritic cells that induces lymphocyte targeting [18]. Chemokines are a family of cytokines that facilitate leukocyte migration and are implicated in the metastatic migration of cancer cells [19]. They are produced not only by immune cells, but also by epithelial and stromal cells [20]. CXCL13 is known to be highly expressed in breast cancer cells [21–25]. CXCL13 mRNAs and proteins are overexpressed in breast cancer cells compared to the expression levels observed in normal breast cells. Furthermore, CXCL13 is the most strongly overexpressed chemokine in breast cancer tissues compared to that in normal breast tissues [24, 25]. The relation between prognosis and triple-negative breast cancer has been controversial, so far. Some authors reported the higher expression of CXCL13 (mRNA/protein) is associated with a lower five-year OS, whereas other author reported the higher expression of CXCL13 mRNA is associated with better distant metastasis-free survival (DMFS) or distant relapse-free survival (DRFS) [25].Chen et al. reported that CXCL13 expression levels were higher in younger women with breast cancer (45 years of age and younger) than that in older women (65 years of age and older). In addition, CXCL13 expression is associated with a high histological grade, lymph node positivity, and ER-negative status [23]. Our findings corroborate the results of previous studies, which have shown that the expression of the chemokine CXCL13 plays a significant role in young patients with breast cancer. Moreover, we have demonstrated that CXCL13 was overexpressed in patients with breast cancer, especially in patients with the luminal A type of PrBC.

Our study was conducted with careful consideration of the microenvironmental differences between the mammary glands of pregnant and lactating women, as well as the differences in gene expression depending on subtype classification. We analyzed gene expression levels using fresh-frozen PrBC tissue samples, excluding PPBC samples, and then validated our findings using microdissected cancer cell components obtained from a large number of FFPE tissue samples. We also examined samples of all subtypes, focusing on the luminal type of PABC. Previous studies in which microarray analysis was conducted using cancer cells have shown that patients with PABC show significantly higher CXCL13 mRNA expression levels than that in patients with non-PABC [26–28]; however, their results were not based on subtype classification, and the studies included patients with PPBC [26]. Breast cancer shows different biological characteristics depending on the subtype; the cancer can be classified into subtypes according to hormone receptor and HER2 receptor expression levels, as well as the Ki-67 labeling index. Although several reports have mentioned that CXCL13 is upregulated or downregulated in patients with PABCs or PrBCs, these controversial results were speculated to be obtained because of the presence of a mixture of different molecular subtypes in the tissue samples. Our study clearly demonstrates that CXCL13 was highly expressed in luminal A-type PrBC.

CXCL13 functions via its receptor C-X-C motif chemokine receptor type 5 (CXCR5). In recent years, it has become clear that CXCL13/CXCR5 signaling plays an important role in the tumor microenvironment of hematological neoplasms. It is known that cancer cells integrate multiple intracellular signaling events and cell-to-cell crosstalk centered on CXCL13/CXCR5 signals and are involved in the acquisition of properties such as invasive growth and metastasis [18]. In this study, microarray analysis revealed that the CXCR5 mRNA expression level was slightly higher in PrBC than that in non-PrBC cases of luminal A-type breast cancer, but the difference was not statistically significant (data not shown). The microarray analysis also showed that no other cytokine-related gene was significantly upregulated in the PrBC group compared to that in the non-PrBC group (data not shown). Functional investigations are required in the future to elucidate the biological mechanisms responsible for overexpression of CXCL13 in luminal A-type PrBC.

Interestingly, the expression level of ER in PrBC cells was observed to be low in this study. A binary estimation of the immunohistochemical properties of the ER protein (that is, positive or negative scores) showed similar results in the PrBC and non-PrBC groups. However, a semi-quantitative scoring system, the Allred score for ER, showed significantly lower scores in the PrBC group than those in the non-PrBC group. Microarray and qRT-PCR analyses of ER mRNA also revealed a low expression level in PrBC cells. The expression levels of ER and PGR in breast cancer tissues are influenced by both age and pregnancy status, and PABCs show a lower ER and PGR positivity rate than that in early-onset breast cancer (EOBC) [2]. One study showed that ER-negative breast cancers were more frequently observed in PrBCs than that in EOBCs [29]. CXCL13 mRNA expression is negatively associated with that of ESR1 in breast cancer cells [30]. In patients with myasthenia gravis, estrogen has been shown to promote CXCL13 secretion during the inflammatory phase and suppress CXCL13 secretion by thymic epithelial cells [31]. It is presumed that both positive and negative correlations between estrogen/ESR1 and CXCL13 expression may occur depending on the presence or the absence of inflammation. In our study, we found an inverse correlation between ESR1 and CXCL13 mRNA expression levels in breast cancer cells. Functional mechanisms related to the microenvironment of cells expressing ER and CXCL13 should be analyzed in future investigations.

To the best of our knowledge, the sample size in the present study was the largest in comparison to the sample sizes reported in studies where genomic analyses of luminal-type PrBCs were conducted. However, a larger number of samples should be examined to obtain clinically significant results.

In conclusion, in this study, we analyzed gene expression in PrBCs and found high expression levels of the chemokine CXCL13, which is involved in immune-related mechanisms, especially in luminal A-type breast cancer that occurred during pregnancy. The limitations of this work include a lack of microarray and qRT-PCR analysis of HER2 and the triple negative type, as well as a lack of protein-level analysis. These results provide clues for a better understanding of breast cancer pathogenesis during pregnancy, which is an immune-tolerant state.

Acknowledgements

The authors thank the staff at the Department of Pathology and Breast Surgical Oncology, St. Luke’s International Hospital. The authors thank Ms. Midori Yokoyama and Ms. Yukari Hirotani for technical support.

Author Contributions

FN: Conceptualization, formal analysis, funding acquisition, investigation, project administration, and writing – original draft. YN: Data curation, formal analysis, and resources. TT: Investigation and resources. TO, RI, YK, KK, JT, AY, and AK: Investigation. NK:Writing – review and editing. HY: Conceptualization and supervision. SM: Conceptualization, supervision, writing – review and editing. All authors read and approved the final manuscript.

Funding

This study was supported by grants from the St. Luke’s Breast Cancer Charity Fund, MEXT KAKENHI (Grant-in-Aid for Young Scientists: grant number 18K15096) and the JBCS, Grant-in-Aid for Establishment of Comprehensive care system for BCP (2017-2019)

Data availability

The microarray data generated in this study are not publicly available because of patient privacy requirements but are available upon request from the corresponding author. The qRT-PCR data in this study are available upon request from the corresponding author.

Ethics declarations

Conflict of interest

The authors declare no potential conflicts of interest.

Ethical approval

The study design was approved by the Institutional Review Board of St. Luke’s International Hospital (approval number 18-R085) according to the Declaration of Helsinki.

Informed consent

Written informed consent was obtained from all participants

Lethaby AE, O’Neill MA, Mason BH, Holdaway IM, Harvey VJ, Auckland Breast Cancer Study Group. Overall survival from breast cancer in women pregnant or lactating at or after diagnosis. Auckland Breast Cancer Study Group. Int J Cancer 1996;67:751–5.
Han SN, Van Calsteren K, Heyns L, Mhallem Gziri M, Amant F. Breast cancer during pregnancy: a literature review. Minerva Ginecol 2010;62:585–97.
Amant F, Lefrère H, Borges VF, Cardonick E, Lambertini M, Loibl S, et al. The definition of pregnancy-associated breast cancer is outdated and should no longer be used. Lancet Oncol 2021;22:753–4.
Albright CM, Wenstrom KD. Malignancies in pregnancy. Best Pract Res Clin Obstet Gynaecol 2016;33:2–18.
Ishida T, Yokoe T, Kasumi F, Sakamoto G, Makita M, Tominaga T, et al. Clinicopathologic characteristics and prognosis of breast cancer patients associated with pregnancy and lactation: Analysis of case-control study in Japan. Jpn J Cancer Res 1992;83:1143–9.
Makita M, Sakamoto G, Namba K, Akiyama F, Iwase T, Sugano H, et al. Study of breast cancers during pregnancy and lactation. Gan No Rinsho 1990;36:39–44. Japanese.
Amant F, von Minckwitz G, Han SN, Bontenbal M, Ring AE, Giermek J, et al. Prognosis of women with primary breast cancer diagnosed during pregnancy: Results from an international collaborative study. J Clin Oncol 2013;31:2532–9.
Johansson AL, Andersson TM, Hsieh CC, Jirström K, Dickman P, Cnattingius S, et al. Stage at diagnosis and mortality in women with pregnancy-associated breast cancer （PABC）. Breast Cancer Res Treat 2013;139:183–92.
Litton JK, Warneke CL, Hahn KM, Palla SL, Kuerer HM, Perkins GH, et al. Case control study of women treated with chemotherapy for breast cancer during pregnancy as compared with nonpregnant patients with breast cancer. Oncologist 2013;18:369–76.
Kataoka A, Ueno T, Yamauchi H, Uehiro N, Takahata C, Takahashi Y, et al. Characteristics, treatment trends, and long-term outcomes of Japanese patients with pregnancy-associated breast cancer (PABC). Breast Cancer 2022;29:825–34.
Korakiti AM, Moutafi M, Zografos E, Dimopoulos MA, Zagouri F. The genomic profile of pregnancy-associated breast cancer: A Systematic Review. Front Oncol 2020;10:1773.
Wolff AC, Hammond MEH, Allison KH, Harvey BE, Mangu PB, Bartlett JMS, et al. Human epidermal growth factor Receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline focused update. J Clin Oncol 2018;36:2105–22.
Coates AS, Winer EP, Goldhirsch A, Gelber RD, Gnant M, Piccart-Gebhart M, et al. Tailoring therapies—Improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015. Ann Oncol 2015;26:1533–46.
Fuchinoue F, Hirotani Y, Nakanishi Y, Yamaguchi H, Nishimaki H, Noda H, et al. Overexpression of PGC1α and accumulation of p62 in apocrine carcinoma of the breast. Pathol Int 2015;65:19–26.
Azim HA Jr, Santoro L, Russell-Edu W, Pentheroudakis G, Pavlidis N, Peccatori FA. Prognosis of pregnancy-associated breast cancer: A meta-analysis of 30 studies. Cancer Treat Rev 2012;38:834–42.
Shao C, Yu Z, Xiao J, Liu L, Hong F, Zhang Y, et al. Prognosis of pregnancy-associated breast cancer: A meta-analysis. BMC Cancer 2020;20:746.
Goddard ET, Bassale S, Schedin T, Jindal S, Johnston J, Cabral E, et al. Association between postpartum breast cancer diagnosis and metastasis and the clinical features underlying risk. JAMA Netw Open 2019;2:e186997.
Hussain M, Adah D, Tariq M, Lu Y, Zhang J, Liu J. CXCL13/CXCR5 signaling axis in cancer. Life Sci 2019;227:175–86.
Luster AD. Chemokines—Chemotactic cytokines that mediate inflammation. N Engl J Med 1998;338:436–45.
Müller A, Homey B, Soto H, Ge N, Catron D, Buchanan ME, et al. Involvement of chemokine receptors in breast cancer metastasis. Nature 2001;410:50–6.
Kazanietz MG, Durando M, Cooke M. CXCL13 and its receptor CXCR5 in cancer: Inflammation, immune response, and beyond. Front Endocrinol (Lausanne) 2019;10:471.
Biswas S, Sengupta S, Roy Chowdhury S, Jana S, Mandal G, Mandal PK, et al. CXCL13-CXCR5 co-expression regulates epithelial to mesenchymal transition of breast cancer cells during lymph node metastasis. Breast Cancer Res Treat 2014;143:265–76.
Chen L, Huang Z, Yao G, Lyu X, Li J, Hu X, et al. The expression of CXCL13 and its relation to unfavorable clinical characteristics in young breast cancer. J Transl Med 2015;13:168.
Panse J, Friedrichs K, Marx A, Hildebrandt Y, Luetkens T, Barrels K, et al. Chemokine CXCL13 is overexpressed in the tumour tissue and in the peripheral blood of breast cancer patients. Br J Cancer 2008;99:930–8.
Jiang L, Wang D, Sheng M, Tong D, Liu H, Dong L, et al. CXCL13/CXCR5 are potential biomarkers for diagnosis and prognosis for breast cancer. J BUON 2020;25:2552–61.
Harvell DM, Kim J, O’Brien J, Tan AC, Borges VF, Schedin P, et al. Genomic signatures of pregnancy-associated breast cancer epithelia and stroma and their regulation by estrogens and progesterone. Horm Cancer 2013;4:140–53.
Azim HA Jr, Brohée S, Peccatori FA, Desmedt C, Loi S, Lambrechts D, et al. Biology of breast cancer during pregnancy using genomic profiling. Endocr Relat Cancer 2014;21:545–54.
Zhang J, Zhou YJ, Yu ZH, Chen AX, Yu Y, Wang X, et al. Identification of core genes and clinical roles in pregnancy-associated breast cancer based on integrated analysis of different microarray profile datasets. Biosci Rep 2019;39:BSR20190019.
Sajjadi E, Venetis K, Noale M, Azim HA Jr, Blundo C, Bonizzi G, et al. Breast cancer during pregnancy as a special type of early-onset breast cancer: Analysis of the tumor immune microenvironment and risk profiles. Cells 2022;11:2286.
Razis E, Kalogeras KT, Kotoula V, Eleftheraki AG, Nikitas N, Kronenwett R, et al. Improved outcome of high-risk early HER2 positive breast cancer with high CXCL13-CXCR5 messenger RNA expression. Clin Breast Cancer 2012;12:183–93.
Pan Z, Zhu T, Liu Y, Zhang N. Role of the CXCL13/CXCR5 axis in autoimmune diseases. Front Immunol 2022;13:850998.

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Breast cancer occurring during pregnancy of Luminal A type overexpressed CXCL13

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Abstract

Background

Methods

Results

Conclusion

Figures

Introduction

Materials and methods

Patients and ethics statement

RNA microarray analysis

RNA extraction and quantitative RT-PCR assays

Statistical analysis

Results

Clinicopathological characteristics of patients with PrBC and non-PrBC

RNA microarray analysis of PrBC and non-PrBC cells of luminal type

Clinicopathological characteristics of patients with luminal A-type PrBC and non-PrBC

Hierarchical cluster analysis of luminal A-type PrBC and non-PrBC samples

Discussion

Declarations

References

Supplementary Files

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