Caffeine ameliorates the metabolic syndrome in diet-induced obese mice through regulating the gut microbiota and serum metabolism.

OBJECTIVE: Obesity is associated with gut microbiota disorders, which has been related to developing metabolic syndromes. The research aims to investigate the effects of caffeine treatment on insulin resistance, intestinal microbiota composition and serum metabolomic changes in high-fat diet (HFD)-induced obesity mice. METHODS: Eight-week-old male C57BL/6 J mice were fed a normal chow diet (NCD) or HFD with or without different concentrations of caffeine. After 12 weeks of treatment, body weight, insulin resistance, serum lipid profiles, gut microbiota and serum metabolomic profiles were assessed. RESULTS: Caffeine intervention improved the metabolic syndrome in HFD-fed mice, such as serum lipid disorders and insulin resistance. 16S rRNA Sequencing analysis revealed that caffeine increased the relative abundance of Dubosiella, Bifidobacterium and Desulfovibrio and decreased that of Bacteroides, Lactobacillus and Lactococcus to reverse HFD-fed obesity in mice. Additionally, Caffeine Supplementation also altered serum metabolomics, mainly focusing on lipid metabolism, bile acid metabolism and energy metabolism. Caffeine increased its metabolite 1,7-Dimethylxanthine, which was positively correlated with Dubosiella. CONCLUSIONS: Caffeine exerts a beneficial effect on insulin resistance in HFD-mice, and the underlying mechanism may be partly related to altered gut microbiota and bile acid metabolism.

TGF-ß1 promotes human breast cancer angiogenesis and malignant behavior by regulating endothelial-mesenchymal transition.

Background: Endothelial-mesenchymal transition (EndMT) is an important process of angiogenesis, which plays a significant role in in tumor invasion and metastasis, while its regulatory mechanisms in breast cancer remain to be fully elucidated. We previously demonstrated that tumor-associated macrophages (TAMs) can induce EndMT in endothelial cells by secreting CCL18 through the activation of the TGF-ß and Notch signaling pathways in breast cancer. This study was designed to study the role of EndMT in breast cancer angiogenesis and progression in order to explore the underlying mechanism. Methods: Immunohistochemistry (IHC) was used to evaluate the expression of microvascular density (MVD) and EndMT markers in breast cancer. TGF-ß1 was used to induce EndMT models of differentiated-endothelial breast cancer stem-like cells (BCSLCs). In vitro cell migration, proliferation and matrigel tube-formation assays, as well as in vivo nude mouse tumor-bearing model and nude mouse dorsal skinfold window chamber (DSWC) model, were utilized to investigate the effects in order to explore the mechanism of EndMT induced by TGF-ß1 on breast cancer progression. Results: In this study, we demonstrated that the EndMT markers were positively associated with MVD indicating unfavorable prognosis of invasive ductal carcinoma (IDC) patients. Functionally, TGF-ß1 promoted migration, proliferation and angiogenesis of differentiated-endothelial BCSLCs by inducing EndMT in vitro and promoted tumor growth and angiogenesis in vivo. Mechanically, we revealed TGF-ß1 induced EndMT by activation of TGF-ß and Notch signaling pathways with increase of p-Smad2/3 and Notch1 expression. Moreover, we found Snail and Slug were key factors of TGF-ß and Notch signaling pathways. Conclusion: Our findings elucidated the mechanism of TGF-ß1 in the promotion of angiogenesis and progression by EndMT in breast cancer.

Corrigendum: TGF-ß1 promotes human breast cancer angiogenesis and malignant behavior by regulating endothelial-mesenchymal transition.

[This corrects the article DOI: 10.3389/fonc.2022.1051148.].

High BTBD7 expression positive is correlated with SLUG-predicted poor prognosis in hormone receptor-negative breast cancer.

BACKGROUND: Hormone receptor-negative breast cancer (HRNBC), which includes triple-negative breast cancer (TNBC) and human epidermal growth factor receptor 2 (HER-2) overexpressing breast cancer, is prone to metastasis and has a poor prognosis. BTB/POZ domain-containing protein 7 (Btbd7) is thought to regulate SLUG and the epithelial-mesenchymal transition (EMT) process. However, the role of Btbd7 in HRNBC is unclear. METHODS: Expression of BTBD7 and SLUG in HRNBC tumor tissue and normal adjacent tissue (NAT) as well as breast cancer cells were characterized by immunohistochemistry and immunofluorescence. MDA-MA-231 cells was transfected with BTBD7 siRNA and detected by qRT-PCR and western blot. Expression levels of Slug and EMT related proteins were detected western blot analysis. cell invasion assays were used to analyse cell invasion ability of MDA-MA-231. GO and KEGG analyses was used to analysis the gene function. RESULTS: The total positive rate of BTBD7 expression in HRNBC tumor tissue was 66.7%, which was higher than that in NAT (52.1%) and benign breast lesion tissues (20%). Co-expression of SLUG and BTBD7 proteins could be found in HRNBC tissue and MDA-MA-231 cells. BTBD7 silencing significantly up-regulated the epithelial marker E-cadherin, down-regulated the mesenchymal markers α-SMA and SLUG and suppressed the invasion abilities of MDA-MA-231 cells. GO and KEGG analyses based on 322 DEGs showed that BTBD7 may be associated with generic transcription in breast cancer. CONCLUSIONS: The study data indicated that BTBD7 was inversely associated with SLUG expression. Higher BTBD7 was associated with poor clinicopathologic features and prognosis in HRNBC patients. BTBD7 silencing inhibited EMT through regulation of SLUG expression. BTBD7 might act as a potential molecular target for gene therapy in HRNBC patients.

Analysis of Lung Adenocarcinoma Subtypes Based on Immune Signatures Identifies Clinical Implications for Cancer Therapy.

Lung cancer is the most common cause of cancer deaths worldwide, and lung adenocarcinoma (LUAD) is the most common histological subtype. However, the prognostic and predictive outcomes differ because of this cancer type heterogeneity. LUAD subtypes were identified on the basis of the immunogenomic profiling of 29 immune signatures. We named three LUAD subtypes: Immunity High, Immunity Medium, and Immunity Low. The Immunity High subtype was characterized by immune activation, e.g., increased immune scores, elevated stromal scores and the highest infiltration of CD8+ T cells, and decreased tumor purities. Activated expressions of human leukocyte antigen (HLA) genes, immune checkpoint molecules, and T helper 1 (Th1)/interferon-gamma (IFNγ) gene signature were also observed in the Immunity High subtype. N 6-methyladenosine (m6A) RNA methylation, associated with cancer initiation and progression, was reduced in the Immunity High subtype. Functional and signaling pathway enrichment analysis further showed that differentially expressed genes between the Immunity High subtype and the other subtypes mainly participated in immune response and some cancer-associated pathways. In addition, the Immunity High subtype exhibited more sensitivity to immunotherapy and chemotherapy. Finally, candidate compounds that aimed at LUAD subtype differentiation were identified. Comprehensively characterizing the LUAD subtypes based on immune signatures may help to provide potential strategies for LUAD treatment.

The prognostic implications of microvascular density and lymphatic vessel density in esophageal squamous cell carcinoma: Comparative analysis between the traditional whole sections and the tissue microarray.

Focal distribution of microvascular and lymphatic vessels is a critical issue in cancer, and is measured by tissue microarray (TMA) construction from paraffin-embedded surgically obtained tissues, a process that may not accurately reflect true focal distribution. The aim of this study was to assess the concordance of microvascular density (MVD) and lymphatic vessel density (LVD) in TMAs with corresponding whole sections, and to correlate the MVD or LVD with clinicopathological parameters in 124 cases of esophageal squamous cell carcinoma (ESCC). MVD, determined by CD105 immunohistochemistry of whole sections, was strongly associated with lymph node metastasis (p=0.000) and pTNM stage (p=0.001). Kaplan-Meier survival analysis showed that increasing CD105 microvessel count correlated with decreasing survival (p<0.001). The same result was acquired when MVD was calculated from tissue microarrays. Analysis of continuous data showed a highly significant correlation between whole sections and TMA data (Pearson r=0.522, p<0.001). Increasing LVD, as determined by D2-40 immunohistochemistry of whole sections, correlated with decreasing survival, but this relationship was undetectable using TMAs. In conclusion, we demonstrate that for the selected endothelial markers, TMAs can provide a realistic and reliable estimate of the extent of MVD, but not LVD in ESCC samples.

Altered expression of endogenous soluble vascular endothelial growth factor receptor-2 is involved in the progression of esophageal squamous cell carcinoma.

Endogenous soluble vascular endothelial growth factor receptor-2 (esVEGFR-2), a new splicing variant of VEGFR-2, was shown to be the first endogenous specific inhibitor of lymphatic vessel growth. The expression of esVEGFR-2 and its clinicopathological roles in esophageal squamous cell carcinoma (ESCC) are unclear. In this article, quantitative RT-PCR was employed to detect the mRNA levels of esVEGFR-2 and VEGF-C in 90 paired primary ESCC tissues, along with immunohistochemical staining to measure esVEGFR-2 protein in 182 ESCC primary tissues. Correlations between esVEGFR-2 expression and clinicopathological features were also analyzed. Compared with the corresponding non-neoplastic esophageal mucosa tissues, the mRNA level of esVEGFR-2 was decreased, whereas the mRNA level of VEGF-C was increased in ESCCs. Downregulation of esVEGFR-2 mRNA level was significantly correlated with pTNM stages (χ(2) = 7.790, p=0.02). Immunohistochemical staining of esVEGFR-2 was inclined to be reduced in ESCC tissues; lower esVEGFR-2 protein expression was related to better prognosis (χ(2) = 6.366, p=0.012), whereas higher esVEGFR-2 protein accumulation in ESCC tissues was an independent prognostic factor for poor survival of patients (hazard ratio, 1.606; 95% confidence interval, 1.042-2.476; p=0.032). Taken together, altered expression of esVEGFR-2 is correlated with progression of ESCC. esVEGFR-2 might serve as a new independent prognostic marker for ESCC patients.

Acoustic radiation force impulse elastography of breast imaging reporting and data system category 4 breast lesions.

INTRODUCTION: This population-based study on early breast cancer detection in women aimed to evaluate acoustic radiation force impulse elastography to differentiate BI-RADS (Breast Imaging Reporting and Data System) category 4 lesions. METHODS: Acoustic radiation force impulse was performed on 95 patients with 122 BI-RADS 4 breast lesions diagnosed by conventional ultrasound. We calculated the area ratio of lesions by using virtual touch tissue imaging and gray-scale imaging. By using virtual touch tissue quantification, we calculated the ratio of shear wave velocity (SWV) in lesions and in surrounding glandular tissue at the same depth (SWV ratio). RESULTS: The mean area ratio of benign lesions (1.09 ± 0.17) differed from that of the malignant lesions (1.96 ± 0.64; P < .001). The mean SWV ratio of benign lesions (2.44 ± 1.27) was lower than that of malignant lesions (5.74 ± 1.68; P < .001). The cutoff for the area and SWV ratios for malignancy were estimated to be 1.37 and 3.65, respectively. CONCLUSIONS: Acoustic radiation force impulse provides quantitative elasticity measurements, which, combined with conventional ultrasound, can potentially improve the diagnostic accuracy of BI-RADS 4 breast lesions, and it is helpful to regulate the BI-RADS classification and avoid the need for unnecessary biopsies.

[Safety and long-term outcome of sleeve lobectomy for non-small cell lung cancer].

BACKGROUND AND OBJECTIVE: Pneumonectomy has been long term used as the standard surgical procedure for central type non-small cell lung cancer (NSCLC). Sleeve lobectomy has been performed in a small number of patients meeting the indications. This study was to compare the 5-year survival rate, operation related complications and mortality of sleeve lobectomy with pneumonectomy for NSCLC, and evaluate sleeve lobectomy in the surgical treatment for NSCLC. METHODS: Ninety-three patients with NSCLC undergoing sleeve lobectomy (group A) and 571 patients with NSCLC undergoing pneumonectomy (group B) from January 1997 to December 2007 in Sun Yat-sen University Cancer Center were reviewed. The 5-year survival rate, operation related complications and mortality between the two groups were analyzed. RESULTS: The overall 5-year survival for group A and group B were 42.0% and 31.5%, respectively (P=0.015). In the subgroup analysis, the 5-year survival of N0 (P=0.007) and N1 (P=0.025) patients were significant higher in group A than in group B, while the survival were not significantly different between N2 patients (P=0.073). The 5-year survival rates for bronchial and pulmonary arterial sleeve resection (the subset of group A) and pneumonectomy were not significantly different (P=0.092). There was no significant difference in local recurrences between the groups (P=0.821). The postoperative complication rates were 11.8% in group A and 20.7% in group B (P=0.046). There was no statistically significant difference in mortality between the two groups (P=0.259). CONCLUSION: The operative safety and long term efficacy of sleeve lobectomy are superior to pneumonectomy for NSCLC.

[Internal mammary node biopsy in breast cancer].

BACKGROUND & OBJECTIVE: Internal mammary node (IMN) is an important lymphatic metastasis pathway in breast cancer. Its status may affect the staging, treatment, prognosis, and outcome evaluation of breast cancer. This study was to discuss the significance of internal mammary sentinel node biopsy (IMSNB), IMN dissection via intercostal spaces, and IMN micro-metastasis detection in breast cancer. METHODS: A total of 38 consecutive patients with primary breast cancer received mastectomy in Cancer Center of Sun Yat-sen University from Dec. 2004 to May 2006. IMSNB was guided by radionuclide tracking method, followed by IMN dissection via the first to forth intercostal spaces. After routine pathologic examination for all the removed IMNs, the negative ones were submitted to further micro-metastasis detection by immunohistochemistry (IHC) combined with multilayer section technique. RESULTS: Of the 38 patients, 17 (44.7%) had internal mammary sentinel nodes (IMSNs). Of the 17 patients, 4 had IMSN metastasis detected by routine pathology, 1 had IMSN micro-metastasis, and 2 had isolated tumor cells in IMSNs, while the remaining 10 had no metastasis. The results of IMSNB were accordant to the results of IMN dissection. Of the 21 (55.3%) patients had no IMSN identified, 5 had IMN metastasis by routine pathology after IMN dissection, and 16 had no metastasis. CONCLUSIONS: When IMSNs are identified, they can predict the IMN status well. While for the patients had no IMSN identified, IMN dissection should be performed, especially for those more likely to have metastasis, to decrease the false negative rate. IHC combined with multilayer section technique tends to search out micro-metastasis.