A case report and a literature review of double mammary pseudoangiomatous stromal hyperplasia associated with galactoma during pregnancy

Pseudoangiomatous stromal hyperplasia (PASH) is a benign interstitial hyperplasia of the breast that usually occurs in premenopausal or perimenopausal women. It is usually characterized by localized lesions or clear boundary masses, and diffuse double breast enlargement is rare. PASH is considered a hormone-dependent disease that is commonly progesterone related. There are no imaging characteristics, and both benign and suspicious malignant signs can be seen. The definitive diagnosis of PASH depends on a pathological diagnosis, and it is necessary to be vigilant in distinguishing between benign and malignant tumors with similar breast histopathology. Here, we report the case of a 23-year-old multipara patient with bilateral diffuse pseudoangiomatous stromal hyperplasia of the breast during pregnancy who presented with macromastia and reviewed the literature to further understand the clinical features, pathological diagnosis, differential diagnosis, treatment and prognosis of pseudoangiomatous stromal hyperplasia of the breast

TGF-β1/Smad Signaling Pathway Regulates Epithelial-to-Mesenchymal Transition in Esophageal Squamous Cell Carcinoma: In Vitro and Clinical Analyses of Cell Lines and Nomadic Kazakh Patients from Northwest Xinjiang, China

<div><p>Invasion and metastasis are the major causes of death in patients with esophageal squamous cell carcinoma (ESCC). Epithelial-mesenchymal transition (EMT) is a critical step in tumor progression and transforming growth factor-β1 (TGF-β1) signaling has been shown to play an important role in EMT. In this study, we investigated how TGF-β1 signaling pathways contributed to EMT in three ESCC cell lines as well as 100 patients of nomadic ethnic Kazakhs residing in northwest Xinjiang Province of China. In vitro analyses included Western blotting to detect the expression of TGF-β1/Smad and EMT-associated proteins in Eca109, EC9706 and KYSE150 cell lines following stimulation with recombinant TGF-β1 and SB431542, a potent inhibitor of ALK5 that also inhibits TGF-β type II receptor. TGF-β-activated Smad2/3 signaling in EMT was significantly upregulated as indicated by mesenchymal markers of N-cadherin and Vimentin, and in the meantime, epithelial marker, E-cadherin, was markedly downregulated. In contrast, SB431542 addition downregulated the expression of N-cadherin and Vimentin, but upregulated the expression of E-cadherin. Moreover, the TGF-β1-induced EMT promoted invasion capability of Eca109 cells. Tumor cells undergoing EMT acquire fibroblastoid-like phenotype. Expressed levels of TGF-β1/Smad signaling molecules and EMT-associated proteins were examined using immunohistochemical analyses in 100 ESCC tissues of Kazakh patients and 58 matched noncancerous adjacent tissues. The results showed that ESCC tissues exhibited upregulated expression of TGF-β1/Smad. We also analyzed the relationship between the above proteins and the patients' clinicopathological characteristics. The TGF-β1/Smad signaling pathway in human Eca109 ESCC cells may carry similar features as in Kazakh ESCC patients, suggesting that TGF-β1/Smad signaling pathway may be involved in the regulation of EMT in ethnic Kazakh patients with ESCC from Xinjiang, China.</p></div

Representative immunohistochemical staining for TGF-β1 (A1,A2), TGF-β RII (B1,B2), p-Smad2/3 (C1,C2), E-cadherin(D1,D2), N-cadherin(E1,E2), and vimentin(F1,F2) in ESCC and NCAT tissues (200×).

<p>Scale bars represent 100 µm.</p

TGF-β1/Smad signaling pathway protein expression in relation to clinical pathological characteristics in Kazakh ESCC.

<p>TGF-β1/Smad signaling pathway protein expression in relation to clinical pathological characteristics in Kazakh ESCC.</p

Transwell assay of Eca109 cells with or without TGF-β1 treatment.

<p>A: Untreated Eca109 cells, 200×. B: Eca109 cells treated with TGF-β1 (10 ng/mL) for 24 h, 200×. C: The number of invading cells: Untreated Eca109 cells vs. Eca109 cells treated with TGFβ1. Data shown represent the average of 3 independent experiments. *<i>p <</i>0.001, compared with untreated cells. All experiments were performed at least 3 times in triplicate. Scale bars represent 100 µm.</p

Western blot analysis of E-cadherin, N-cadherin, vimentin, P-Smad2 protein expression in Eca109(2A–D), Eca9706(2E–H), KYSE150(2I–L) cells treated with TGF-β1 or SB431542 at differential concentrations.

<p><b>A</b>. Effects of treatment of Eca109 cells with TGF-β1 (1, 5, or 10 ng/mL) on the expression of E-cadherin (molecular weight 97 kDa), N-cadherin (molecular weight 100 kDa), vimentin (molecular weight 57 kDa), p-Smad2 (molecular weight 52 kDa) and Smad7 (molecular weight 51 kDa) by Western Blot. <b>B</b>. Quantiative analysis of treatment of Eca109 cells with TGF-β1 (1, 5, or 10 ng/mL), E-cadherin, N-cadherin, vimentin, P-Smad2 and Smad7 expression levels; Y axis: banding densities of test marker versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01. <b>C</b>. Effects of SB431542 (1, 5, or 10 µM) on the expression of E-cadherin, N-cadherin, vimentin, Smad2/3 and Smad7 in Eca109 cells by Western Blot. <b>D</b>. Quantiative analysis of treatment of Eca109 cells with SB431542 (1, 5, or 10 µM), E-cadherin, vimentin, Smad2/3 and Smad7 expression levels; Y axis: banding densties of test versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01; Each experiment was repeated three times. <b>E</b>. Effects of treatment of Eca 9706 cells with TGF-β1 (1, 5, or 10 ng/mL) on the expression of E-cadherin, N-cadherin, vimentin, p-Smad2 by Western Blot. <b>F</b>. Quantiative analysis of treatment of Eca 9706 cells with TGF-β1 (1, 5, or 10 ng/mL), E-cadherin, N-cadherin, vimentin, P-Smad2 expression levels; Y axis: banding densities of test marker versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01. <b>G</b>. Effects of SB431542 (1, 5, or 10 µM) on the expression of E-cadherin, N-cadherin, vimentin, Smad2/3 in Eca 9706 cells by Western Blot. <b>H</b>. Quantiative analysis of treatment of Eca 9706 cells with SB431542 (1, 5, or 10 µM), E-cadherin, vimentin, Smad2/3 expression levels; Y axis: banding densties of test versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01; Each experiment was repeated three times. <b>I</b>. Effects of treatment of KYSE150 cells with TGF-β1 (1, 5, or 10 ng/mL) on the expression of E-cadherin, N-cadherin, vimentin, p-Smad2 by Western Blot. <b>J</b>. Quantiative analysis of treatment of KYSE150 cells with TGF-β1 (1, 5, or 10 ng/mL), E-cadherin, N-cadherin, vimentin, P-Smad2 expression levels; Y axis: banding densities of test marker versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01. <b>K</b>. Effects of SB431542 (1, 5, or 10 µM) on the expression of E-cadherin, N-cadherin, vimentin, Smad2/3 in KYSE150 cells by Western Blot. <b>L</b>. Quantiative analysis of treatment of KYSE150 cells with SB431542 (1, 5, or 10 µM), E-cadherin, vimentin, Smad2/3 expression levels; Y axis: banding densties of test versus β-actin. Data are expressed as a significant change relative to the control. Each bar represents the mean±s.d. *, p<0.05, **, p<0.01. Each experiment was repeated three times.</p

Morphological changes of Eca109(1–1) and KYSE150(1–2) cells after TGF-β1 treatment.

<p>A: Untreated Eca109 and KYSE150 cells, 100×; B,C: Eca109 and KYSE150 cells treated with TGF-β1 (1,5 ng/mL) for 36 hrs, 100×. D Control stimulation with TGF-β1 (5 ng/mL) in the presence of SB4315425(5 µM) for 36 hrs, 100×. Scale bars represent 50 µm.</p

Correlation between the expression of E-cadherin protein and N-cadherin/vimentin proteins in ESCC.

<p><i>r</i> = −0.231, <i>p</i> = 0.021 vs. N-cadherin; r = −0.055, <i>p</i> = 0.590 vs. vimentin.</p><p>Correlation between the expression of E-cadherin protein and N-cadherin/vimentin proteins in ESCC.</p

Correlation between the expression of TGF-β1 protein and TGF-βRII/p-Smad2/3 proteins in ESCC.

<p>Correlation between the expression of TGF-β1 protein and TGF-βRII/p-Smad2/3 proteins in ESCC.</p

The comparison between the tumor and the adjacent tissue of the same patients.

<p>The comparison between the tumor and the adjacent tissue of the same patients.</p