A novel approach for transforming breast cancer stem cells into endothelial cells.

Tumor vascular endothelial cells play a pivotal in the tumor microenvironment, influencing the proliferation, invasion, and metastasis of tumor progression. The present study investigated a novel method for inducing the transformation of breast cancer stem cells into endothelial cells, providing a cellular model investigating anti-angiogenic mechanisms in vitro. The breast cancer cell line MCF-7 was used, and the expression of CD133 was initially detected using flow cytometry. CD133+ breast cancer cells were purified using immunomagnetic bead sorting technology, yielding an MCF-7CD133+ subpopulation. The proliferation ability of these cells was assessed using an MTT assay, while their microsphere formation ability was evaluated using a microsphere formation assay. Post-transformation in an optimized endothelial cell culture medium, expression of endothelial cell markers CD31 and CD105 were detected using flow cytometry. Endothelial cell tube formation assays and DiI-labeled acetylated low-density lipoprotein (DiI-Ac-LDL) assays were employed to analyze the endothelial cell function of the MCF-7CD133+ cells. MDM2/CEN12 gene amplification was detected through fluorescence in situ hybridization (FISH). The MCF-7 breast cancer cell line exhibited 1.7±0.3% trace cells expressing the stem cell surface marker CD133. After anti-CD133 immunomagnetic bead sorting, MCF-7CD133+ and MCF-7CD133- subpopulation cells were obtained, with CD133 expression rates of 85.6±2.8 and 0.18±0.08%, respectively. MTT assay results demonstrated that, after 7 days, the proliferation rate of MCF-7CD133+ cells was significantly higher compared with MCF-7CD133- cells. MCF-7CD133+ subpopulation cells displayed strong stem cell characteristics, growing in suspension in serum-free media and forming tumor cell spheres. In contrast, MCF-7CD133- cells failed to form microspheres. After culturing cells in endothelial cell differentiation and maintenance media, the percentage of MCF-7CD133+ cells before and after endothelial cell culture was 0.3±0.16 and 81.4±8.37% for CD31+ cells and 0.2±0.08 and 83.8±7.24% for CD105+ cells, respectively. Vascular-like structure formation and Ac-LDL phagocytosis with red fluorescence in the tube formation assays confirmed endothelial cell function in the MCF-7CD133+ cells. FISH was used to verify MDM2/CEN12 gene amplification in the induced MCF-7CD133+ cells, indicating tumor cell characteristics. The modified endothelial cell transformation medium effectively induced differentiated tumor stem cells to express vascular endothelial cell markers and exhibit endothelial functions, ideal for in vitro anti-angiogenesis research.

Retracted: Microcalcification-associated breast cancer: HER2-enriched molecular subtype is associated with mammographic features.

OBJECTIVE: To investigate whether the mammographic features were different between breast cancer HER2-enriched molecular subtype and non-HER2-enriched molecular subtype. METHODS: 283 microcalcification-associated breast cancers were identified (HER2-enriched: n = 57; non-HER2-enriched: n = 226). Mammographic tumor mass and calcification features in relation to HER2 molecular subtype were analyzed. RESULTS: On univariate analysis, HER2-enriched molecular subtype rates were significantly higher (a) in tumor size ≤2 cm [33 of 57 (57.9%)] than in tumor size >2 cm lesions [22 of 226 (9.7%)] (p = 0.007), (b) in non-spiculated mass [39 of 57 (68.4%)] than in spiculated mass lesions [18 of 226 (7.9%)] (p = 0.034)，(c) in calcifications extent >2 cm [41 of 57 (71.9%)] lesions than in calcifications extent ≤2 cm lesions [16 of 226 (7.1%)] (p < 0.001) and (d) in calcification density ＞20 cm-2 [44 of 57 (71.2%)] lesions than in calcification density ≤20 cm-2 lesions [13 of 226 (5.8%)] (p = 0.034).On multivariate analysis, three mammographic features [tumor size >2 cm vs size ≤2 cm odds ratio (OR): 0.415 95% confidence interval (CI) (0.215 to 0.802), p = 0.009, spiculated mass vs non-spiculated mass OR: 0.226 95% CI (0.114 to 0.446), p < 0.001 and calcifications extent >2 cm vs calcifications extent ≤2 cm OR: 7.754, 95% CI (3.100 to 19.339) p< 0.001] were independent predictors. Our results indicated that small tumor size, non-spiculated mass and calcification extent >2 cm are more likely to be HER2 molecular subtype. The discrimination of this model, as quantified by the area under the curve, was 0.751 [95% CI (0.701 to 0.854)]. CONCLUSION: Our study presents a prediction model that incorporates the mammographic features of tumor size, non-spiculated mass and calcification extent, which can potentially be used to preoperative predict breast cancer HER2-enriched subtype. ADVANCES IN KNOWLEDGE: Mammographic features can noninvasively visualize breast tumor phenotype characteristics.

Serum levels of 25-hydroxyvitamin D predicts infarct volume and mortality in ischemic stroke patients.

BACKGROUND: The aim of this study is to determine 25-hydroxyvitamin D [25(OH) D] levels in serum, and investigate their associations with cardiovascular disease (CVD) or all-cause mortality in a 1-year follow-up study in patients with first-ever ischemic stroke. METHODS: From November 2013 to October 2015, 387 consecutive patients with ischemic stroke admitted to our hospital were identified. Serum 25(OH) D levels were measured at admission. Infarct volume was measured using diffusion-weighted imaging (DWI). The primary end point was CVD mortality among 1year. The secondary end point was all-cause mortality. RESULTS: In this study, 387 patients were included. A statistically significant negative correlation between serum 25(OH) D level and infarct volume was found (r=-0.442; P<0.001). There were 74 patients (19.1%, 95%CI: 15.2%-23.0%) died, including 36 CVD mortality (9.3%, 95CI%: 6.4%-12.2%). The mortality distribution across the 25(OH) D quartiles ranged between 39.2% (first quartile) to 5.2% (fourth quartile) for all-cause mortality and between 18.6% (first quartile) to 2.1% (fourth quartile) for CVD mortality. In a multivariate model using the first quartiles of 25(OH) D vs. quartiles 2 through 4 together with the clinical variables, the marker displayed prognostic information CVD mortality: OR for first quartile, 3.06 [95% CI, 2.16-4.95]; all-cause mortality: OR for first quartile, 2.76 [95% CI, 2.01-4.32]. CONCLUSIONS: The data show serum levels of 25(OH) D at admission is useful prognostic marker of CVD and all-cause mortality in Chinese patients with ischemic stroke.