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.

FDX1 inhibits thyroid cancer malignant progression by inducing cuprotosis.

Cuprotosis is a recently identified cell death form that caused by intracellular copper accumulation and regulated by FDX1. This work aimed to explore the role of cuprotosis and the pivotal regulatory gene FDX1 in thyroid cancer development. We observed that expression of FDX1 in tumor section was notably lower than that in non-tumor sections in clinical samples. Induction of cuprotosis by elesclomol (ES) significantly repressed the in vitro and in vivo growth of thyroid cancer cells, simultaneously elevated Cu level and expression of FDX1, whereas depletion of FDX1 abolished these effects. Knockdown of FDX1 decreased the lipoylation level of DLAT and DLST in thyroid cancer cells, alleviated cuprotosis-induced cell death, simultaneously upregulated the levels of PA and α-KG. These findings demonstrated that FDX1 promotes the cuprotosis of thyroid cancer cells via regulating the lipoylation of DLAT.

"Elastic stretch cavity building" system in endoscopic thyroidectomy via the axillary approach: a case series.

Background: Patients undergoing conventional endoscopic thyroidectomy via the axillary approach, which is commonly used clinically, suffered from a range of postoperative complications. This study aimed to prevent postoperative complications and evaluate patients' satisfaction with cosmetic outcomes in endoscopic thyroidectomy via the axillary with the use of "Elastic Stretch Cavity Building" System. Methods: In this retrospective case series study, the clinical data of patients who were admitted to the Thyroid Surgery Department of Ningbo Medical Centre Lihuili Hospital between December 2020 and December 2021 for endoscopic thyroidectomy via the axillary approach under the "Elastic Stretch Cavity Building" System. Results: A total of 67 patients were included, all surgeries were successfully completed. The operation time was 75.61 ± 13.67 minutes; the postoperative drainage volume was 109.97 ± 37.54 ml; the average postoperative hospital stay was 4 (2-6) days. There was no skin ecchymosis, effusion or infection, hypocalcemia, convulsions, upper extremity dyskinesia, and temporary hoarseness after the surgery. The patients were satisfied with the cosmetic effects, and the cosmetic score was 4 (3-4). Conclusion: The "Elastic Stretch Cavity Building" System in endoscopic thyroid surgery via the axillary approach might reduce the risks of complications and achieve satisfactory results with the cosmetic outcomes.

SBP-0636457, a Novel Smac Mimetic, Cooperates with Doxorubicin to Induce Necroptosis in Breast Cancer Cells during Apoptosis Blockage.

Breast cancer (BC) is a common health concern worldwide. Doxorubicin (Dox) is a widely used chemotherapeutic agent to treat various cancers, including BC. However, drug resistance and severe side effects often hinder the clinical application of Dox. Combination therapy is an effective potent strategy to increase chemosensitivity and reduce the adverse effects. Smac is a proapoptotic protein that interacts with inhibitors of apoptosis proteins (IAPs) and thereby promotes cell death. Smac mimetic compounds can mimic its function and can be used to kill cancer cells. In this study, Dox and SBP-0636457, a novel Smac mimetic, were found to have cooperative effects in inducing BC cell death. Dox and SBP-0636457 cotreatment induced necroptosis instead of apoptosis in BC cells. Receptor-interacting serine/threonine-protein kinase 1 or mixed-lineage kinase domain-like silencing could attenuate cell death caused by Dox/SBP-0636457 in BC cells. In addition, this combined treatment caused synergistic induction of TNFα, and TNFα/TNFR signalling is essential for cell death induced by Dox/SBP-0636457 in BC cells. Moreover, both canonical and noncanonical nuclear factor kappa B pathways were found to contribute to TNFα upregulation induced by Dox/SBP-0636457. Therefore, the findings suggest that SBP-0636457 combined with Dox is an alternative strategy for treating BC.

"Elastic Stretch Cavity Building" System in Endoscopic Thyroidectomy of Giant Thyroid Tumors.

Objective: To analyze the clinical characteristics of patients with large thyroid tumors underwent endoscopic thyroidectomy using the "elastic stretch cavity builder" system. Methods: This retrospective case series study included thyroid tumor patients admitted to the Ningbo Medical Center Li Hui li Hospital between September 2017 and November 2021. The self-developed "elastic stretch cavity builder" was used to elastically lift the anterior cervical flap, combined with low-pressure (3 mmHg) high-flow CO2 inflation, and create a working cavity for endoscopic thyroidectomy. Results: This study included 13 patients for analysis. The endoscopic thyroidectomy duration was 92-170 min (mean, 123 ± 24min). The maximum transverse plane diameter of the glands was 5.0-6.2 cm (mean, 5.3 ± 0.3 cm). The maximum sagittal plane diameter was 6.8-10.0 cm (mean, 7.6 ± 0.9 cm). After the "elastic stretch cavity builder" lifted the cervical flap, the height of the subcutaneous region was increased by 1.3 ± 0.2cm without affecting cervical activity. There was no residual scar in the anterior cervical skin puncture hole. All patients were satisfied with the cosmetic with the cosmetic satisfaction score was 3.4 ± 0.5. Conclusion: The novel mixed cavity building model established by the "elastic stretch cavity builder" might provide the surgeon with additional longitudinal cervical operating space while improving the stability of the space and saving human effort.

BAY-885, a mitogen-activated protein kinase kinase 5 inhibitor, induces apoptosis by regulating the endoplasmic reticulum stress/Mcl-1/Bim pathway in breast cancer cells.

The mitogen-activated protein kinase kinase 5 (MEK5)/extracellular signal-regulated kinase 5 (ERK5) axis has been reported to promote tumorigenesis in breast cancer (BC). Therefore, targeting the MEK5/ERK5 axis is a potential strategy against BC. BAY-885 is a novel inhibitor of ERK5; however, to date, its anti-tumor effects in BC have not been investigated. This study aimed to assess the anti-tumor effects of BAY-885 in BC and identify its underlying mechanisms of action. Unlike other ERK5 inhibitors, which frequently failed to mimic ERK5 genetic ablation phenotypes, the BAY-885 treatment effectively recapitulated ERK5 depletion effects in BC cells. Results revealed that BAY-885 affected the viability and induced apoptosis in BC cells. Moreover, the BAY-885-mediated downregulation of myeloid cell leukemia-1 (Mcl-1) and upregulation of Bim were dependent on ERK5 inhibition. Furthermore, BAY-885 triggered activation of endoplasmic reticulum (ER) stress, which further led to the upregulation of Bim and downregulation of Mcl-1. ER stress was induced in an ERK5 inhibition-dependent manner. These findings suggested that BAY-885 induced apoptosis in BC cells via ER stress/Mcl-1/Bim axis, suggesting that BAY-885 may serve as a therapeutic agent for BC.

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.

Hypermethylated KLF9 Is An Independent Prognostic Factor For Favorable Outcome In Breast Cancer.

BACKGROUND AND OBJECTIVE: Breast cancer (BC) is the most lethal human malignancy and is the leading cause of cancer-associated death in women worldwide. Krüppel-like factor 9 (KLF9) belongs to a family of transcriptional regulators and its role in BC has not been fully investigated. METHOD: Data mining was used to analyze BC data from The Cancer Genome Atlas (TCGA) database, which was downloaded using the UCSC Xena browser. The differential expression and methylation level of KLF9 was analyzed in patients with BC and corresponding normal controls enrolled from our hospital. Besides, the correlation of KLF9 methylation and prognosis was explored, and gene set enrichment analysis (GSEA) was conducted to identify the potential signaling pathway of KLF9 involved. RESULTS: Both TCGA and BC tissues indicated hypermethylation of the KLF9 promoter region in patients with BC compared with normal controls, which might account for the dysregulation of KLF9 in patients with BC. Besides, hypermethylation of KLF9 was detected in patients with estrogen or progesterone receptor-positive and non-triple-negative disease. Further, hypermethylation of KLF9 was demonstrated to be a potential independent biomarker in obtaining favorable outcomes in BC. By GSEA, tumor-associated biological processes and signaling pathway were identified, which indicated that KLF9 might play a vital role in the carcinogenesis of BC. CONCLUSION: KFL9 plays an important role in the carcinogenesis of BC through the multiple tumor-associated signaling pathway. The hypermethylation of KLF9 resulted in its reduced expression in BC, while the hypermethylation of KLF9 has potential in the prediction of favorable outcomes in BC.

Silencing of rhomboid domain containing 1 to inhibit the metastasis of human breast cancer cells in vitro.

OBJECTIVES: A growing body of evidence indicates that rhomboid domain containing 1 (RHBDD1) plays an important role in a variety of physiological and pathological processes, including tumorigenesis. We aimed to determine the function of RHBDD1 in breast cancer cells. MATERIALS AND METHODS: In this study, we used the Oncomine™ database to determine the expression patterns of RHBDD1 in normal and breast cancer tissues. We performed lentiviral transfection of RHBDD1-specific small interfering RNA into the breast cancer cell lines ZR-75-30 and MDA-MB-231 in order to investigate the effects of RHBDD1 deficiency on breast cancer metastasis. RESULTS: We found that knockdown of RHBDD1 inhibited breast cancer cell migration and invasion in vitro. Moreover, knockdown of RHBDD1 promoted epithelial-mesenchymal transition (EMT) by suppressing the expression of MPP2, MPP9, fibronectin 1, vimentin, SRY-box 2, zinc finger E-box binding homeobox 1, and snail family transcriptional repressor 1, and promoting the expression of cadherin 1. Additionally, knockdown of RHBDD1 inhibited the protein expression and phosphorylation of Akt. CONCLUSION: Our data indicate that RHBDD1 overexpression may promote breast cancer metastasis via the regulation of EMT, suggesting that RHBDD1 may be an important regulator of breast cancer metastasis.

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.