MicroRNAs (miRs/miRNAs) play important roles in the occurrence, metastasis and prognosis of multiple types of cancers. However, the specific role of miR-302d-3p and its underlying mechanism in breast cancer (BC) have not yet been reported. The present study aimed to identify the role of miR-302D-3p in BC and its potential mechanism using BC cell lines MCF7 and MDA-MB-231 and normal breast epithelial cell MCF-10A. Cancer and paracancerous tissue from patients with BC were also used. Reverse transcription-quantitative PCR was performed to detect the expression of miR-302d-3p and transmembrane Bax inhibitor motif containing 6 (TMBIM6). Dual-luciferase reporter assays verified the binding sites of miR-302d-3p and TMBIM6. Immunohistochemistry was used to measure the expression of TMBIM6. Cell transfection techniques were used to overexpress or interfere with miR-302d-3p and TMBIM6. A Cell Counting Kit-8 assay was performed to detect cell viability, and migration was measured using a wound healing assay. Apoptosis was detected by flow cytometry. The expression levels of apoptosis-related proteins and pathway-related proteins were detected by western blotting. The expression of miR-302d-3p in BC cell lines was found to be downregulated. It was also demonstrated that miR-302d-3p could inhibit cell viability and migration and promote apoptosis. The expression of TMBIM6 in BC cell lines and tissues was upregulated. Upregulated miR-302d-3p was shown to inhibit viability and migration, and promote apoptosis by targeting TMBIM6, during which extracellular signal-regulated kinase (ERK) and its phosphorylation were inhibited in the ERK signaling pathway in cells. Overall, the present study demonstrated that miR-302d-3p could regulate the viability, migration and apoptosis of BC cells through regulating TMBIM6-mediated ERK signaling pathway.

Background Traditional phenotype-based screening for β-globin variant and β-thalassemia using hematological parameters is time-consuming with low-resolution detection. Development of a MALDI–TOF–MS assay using alternative markers is needed. Methods We constructed a MALDI–TOF–MS-based approach for identifying various β-globin disorders and classifying thalassemia major (TM) and thalassemia intermedia (TI) patients using 901 training samples with known HBB/HBA genotypes. We then validated the accuracy of population screening and clinical classification in 2 separate cohorts consisting of 16 172 participants and 201 β-thalassemia patients. Traditional methods were used as controls. Genetic tests were considered the gold standard for testing positive specimens. Results We established a prediction model for identifying different forms of β-globin disorders in a single MALDI–TOF–MS test based on δ- to β-globin, γ- to α-globin, γ- to β-globin ratios, and/or the abnormal globin-chain patterns. Our validation study yielded comparable results of clinical specificity (99.89% vs 99.71%), and accuracy (99.78% vs 99.16%) between the new assay and traditional methods but higher clinical sensitivity for the new method (97.52% vs 88.01%). The new assay identified 22 additional abnormal hemoglobins in 69 individuals including 9 novel ones, and accurately screened for 9 carriers of deletional hereditary persistence of fetal hemoglobin or δβ-thalassemia. TM and TI were well classified in 178 samples out of 201 β-thalassemia patients. Conclusions MALDI–TOF–MS is a highly accurate, predictive tool that could be suitable for large-scale screening and clinical classification of β-globin disorders.

Background: High-fat, low-carbohydrate and adequate-protein ketogenic diets (KD) are expected to become an effective adjunct therapy for cancer patients. However, the direct effects of KD on tumor cells and the underlying mechanisms are elusive. In this study, the nude mouse models of subcutaneous transplanted human colon cancer cells were established and applied to study the effects and mechanisms of KD on the growth of subcutaneous tumors in nude mice.Methods: Male nude mice were injected subcutaneously with human colon cancer HCT-116 cell line to construct a subcutaneous tumor model of human colon cancer. The successfully constructed subcutaneous tumor mice were divided into normal diet group and KD group. The mice were anesthetized and euthanized after 30 days of feeding, the subcutaneous tumor tissues were collected, and the size of tumors was measured and weighed. HE staining was used to observe the pathological changes of subcutaneous tumor tissues in normal feeding group and KD group. TUNEL staining was used to detect the level of apoptosis in tumor tissue. Immunohistochemistry of subcutaneous tumor tissues was used to detect the expression levels of Wnt-1 and β-catenin. In addition, RT-qPCR and western blotting were applied to detect the expression levels of Wnt1/β-catenin signaling pathway-related proteins.Results: After 30 days of normal diet and KD feeding, the subcutaneous tumor tissues of human colon cancer mice were taken out for various assays. The results of tumor size measurement showed that the tumor size and weight of KD group were significantly smaller than that of the normal diet group. HE staining showed that the pathological characteristics of colon tumor tissue in the KD group were significantly improved, and the infiltration of inflammatory cells was reduced. TUNEL staining showed that the apoptosis level of tumor cells in the KD group was significantly increased compared to the normal diet group. RT-qPCR and western blotting revealed that the expression of pro-apoptotic proteins such as caspase 3，caspase 9 and Bax were increased（P < 0.01）, while the expression of anti-apoptotic protein such as Bcl-2 or survivin was decreased （P < 0.01）. Furthermore, the expression of Wnt1/β-catenin signaling pathway-related proteins including Wnt1 and β-catenin were largely reduced after 30 days of KD feeding compared to normal feeding group （P < 0.01）.Conclusions: Ketogenic diets (KD) promotes apoptosis of human colon cancer subcutaneous tumor cells and inhibits the growth of tumor by inhibiting Wnt1/β-catenin signaling pathway in mouse subcutaneous tumor models of human colon cancer.