MAC mediates mammary duct epithelial cell injury in plasma cell mastitis and granulomatous mastitis.

Plasma cell mastitis (PCM) and granulomatous mastitis (GM) are common inflammatory nonbacterial mastitis (NBM). However, the pathogenesis of NBM is still unclear. METHODS: In this study, we statistically analyzed the pathological features of PCM and GM using pathological HE staining and tissue transmission electron microscopy. The levels of MAC (C5b-9n), P-selectin, E-selectin, and ICAM-1 were detected through IHC, WB, ELISA, and qPCR. The expression level and location of MAC were observed by tissue immunological electron microscopy. In addition, exosomes were isolated from tissues, identified using transmission electron microscopy, and the densities were detected by Nano-FCM. Finally, the expression intensity of MAC in exosomes was detected by flow cytometry and immunoelectron microscopy. RESULTS: The damage and apoptosis of mammary duct epithelial cells are the common pathological features of PCM and GM. MAC is primarily located in the cell membrane of mammary ductal epithelial cells and is significantly expressed in PCM and GM. The density of exosomes in PCM and GM tissues was elevated, and MAC was highly expressed in exosomes. In addition, the expression of P-selectin, E-selectin, and ICAM-1 in PCM and GM was significantly higher than in the normal group. CONCLUSION: We found severe damage of the mammary duct epithelial cells in PCM and GM tissues, which was verified by relevant pathological methods. Earlier studies demonstrated that MAC is highly expressed in PCM and GM tissues and exosomes seem to play a very important role in the understanding of MAC. Furthermore, MAC is involved in inflammatory infiltration and lesion of mammary duct epithelial cells upregulated by P-selectin, E-selectin, and ICAM-1. These findings provide new insights into PCM and GM molecular mechanisms.

LncRNA AFAP1-AS1 Knockdown Represses Cell Proliferation, Migration, and Induced Apoptosis in Breast Cancer by Downregulating SEPT2 Via Sponging miR-497-5p.

Background: Long non-coding RNA actin filament-associated protein1-antisense RNA 1 (AFAP1-AS1) was confirmed to be associated with tumorigenesis. However, the role of AFAP1-AS1 in breast cancer was little known. Materials and Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the levels of AFAP1-AS1, microRNA-497-5p (miR-497-5p), and Septin 2 (SEPT2) in breast cancer tissues and cells. The cell proliferation, migration, and apoptosis were tested by Methylthiazolyldiphenyl-tetrazolium bromide (MTT), Transwell and Flow cytometry assays, respectively. The targeting relationship between genes was predicted by StarBase v.3.0 and confirmed by dual-luciferase reporter assay. Pearson's correlation coefficient was applied to examine the correlation between the two groups. SEPT2 protein expression was evaluated by Western blot. Xenograft models were established to investigate the role of AFAP1-AS1 knockdown in vivo. Results: AFAP1-AS1 was upregulated in breast cancer tissues and cells, and AFAP1-AS1 knockdown could hinder proliferation and migration of breast cancer cells, and contribute to cell apoptosis. MiR-497-5p, which was downregulated in breast cancer, was verified to be a target of AFAP1-AS1 and inversely correlated with AFAP1-AS1 expression. SEPT2, as a target gene of miR-497-5p, was negatively regulated by miR-497-5p and positively correlated with AFAP1-AS1 expression. Importantly, AFAP1-AS1 could upregulate SEPT2 expression by sponging miR-497-5p, and modulate cell progression by regulation of the miR-497-5p/SEPT2 axis in breast cancer. Conclusion: AFAP1-AS1 knockdown repressed the progression of breast cancer cells by sponging miR-497-5p and downregulating SEPT2.

Composition and plasticity of triple-negative breast carcinoma-infiltrating regulatory T cells.

Low Foxp3+ regulatory T-cell (Treg) presence in the tumor-infiltrating lymphocytes (TILs) is considered favorable in breast cancer, and numerous CD25-targeting agents have been applied in the attempt to remove Foxp3+ Treg cells, which typically present CD4+ CD25+/hi surface phenotype. However, CD25 is not Treg-exclusive and can be upregulated by effector T cells. Hence, CD25 depletion may cause the elimination of activated T cells that are responding to tumor-specific antigens. In this study, the composition and function of CD4+ CD25+ cells inside the microenvironment of triple-negative breast carcinoma (TNBC) were investigated. Directly ex vivo, the Foxp3+ Treg cells represented a minor subset in total CD4+ CD25+ TILs. Significant differences were observed in the expression of Treg-associated molecules between CD4+ CD25+ Foxp3+ TILs and CD4+ CD25+ Foxp3- TILs. While both the CD4+ CD25+ Foxp3+ and the CD4+ CD25+ Foxp3- TILs could express CTLA-4 and LAG-3, the expression levels were significantly higher in CD4+ CD25+ Foxp3+ TILs than in CD4+ CD25+ Foxp3- TILs. Upon TCR stimulation, the expression of TGF-beta was significantly higher in CD4+ CD25+ Foxp3+ TILs, while the expression of IL-10 was significantly higher in CD4+ CD25+ Foxp3- TILs. These differences were conserved in the blood counterparts of these cells. Interestingly, the level of CD25+ Foxp3+ cells in circulating CD4+ T cells was positively correlated with the level of CD25+ Foxp3+ cells in CD4+ TILs, but the level of CD25+ Foxp3- cells in circulating CD4+ T cells was not associated with the level of CD25+ Foxp3- cells in CD4+ TILs. Th17-polarizing medium could readily remodel CD4+ CD25+ Foxp3- , but not CD4+ CD25+ Foxp3+ , T cells into RORgammat and IL-17-expressing T cells, demonstrating stronger plasticity of the former subset. Together, these data demonstrated that the CD4+ CD25+ TILs were composed of distinctive Foxp3- and Foxp3+ cells, with the former representing the major subset. The antigen specificity and effector molecule expression of the CD4+ CD25+ Foxp3- thus require further analyses.

IL-9-producing CD8+ T cells represent a distinctive subset with different transcriptional characteristics from conventional CD8+ T cells, and partially infiltrate breast tumors.

Accumulating evidence suggests that IL-9 and IL-9-producing cells exert various roles in antitumor immunity. Our study examined the IL-9 production in CD8+ T cells from breast cancer patients as compared to healthy controls. IL-9 secretion was undetectable in CD8+ T cells ex vivo, but could be readily detected following anti-TCR or PMA + ionomycin stimulation, and was higher in breast cancer patients than in healthy controls. The capacity to express IL-9 was not universal to all CD8+ T cells, but was favored in IL-9Rhigh CD8+ T cells, which were also present in breast cancer patients at significantly higher frequency than in healthy controls. Interestingly, exogenous IL-9 could significantly increase the expression of both IL-9 and IL-9R in IL-9Rhigh, but not IL-9Rlow, CD8+ T cells. IL-9Rhigh CD8+ T cells ex vivo presented lower expression of KLRG-1, PD-1, and Tim-3 than IL-9Rlow CD8+ T cells. Additionally, IL-9Rhigh CD8+ T cells following anti-TCR and PMA + ionomycin stimulation presented higher IL-2 and IL-17 expression, and lower IFN-γ expression, than IL-9Rlow CD8+ T cells. IL-9-expressing CD8+ T cells could be found in some, but not all, resected breast tumors. IL-9R expression, on the other hand, was readily present in CD8+ T cells, but with high variability from patient to patient. Patients with high intratumoral IL-9 expression also tended to present high IL-9R expression. Together, these data demonstrate that a transcriptionally distinctive IL-9-producing CD8+ T cell subset was elevated in breast cancer patients and could be found inside the tumor, with higher capacity to produce IL-2 and IL-17 and lower expression of inhibitory receptors.

Osteopontin versus alpha-fetoprotein as a diagnostic marker for hepatocellular carcinoma: a meta-analysis.

BACKGROUND: Several studies have reported that osteopontin (OPN) is a promising marker for the diagnosis of hepatocellular carcinoma (HCC); however, some studies emerged with conflicting results. Therefore, we provide a systematic review to evaluate the diagnostic performance of OPN for HCC. METHODS: Studies that investigated the diagnostic value of OPN and alpha-fetoprotein (AFP) in HCC were collected from PubMed and Embase. Sensitivity, specificity, and other parameters about the diagnostic accuracy of serum OPN and AFP in HCC were pooled using STATA 12.0 software. The summary receiver operating characteristic curve (sROC) and other parameters were used to summarize the overall test performance. RESULTS: Twelve studies were included in our meta-analysis. Pooled sensitivity, specificity, and diagnostic odds ratio were 0.813 (95% CI: 0.671-0.902), 0.874 (95% CI: 0.778-0.932), and 30.047 (95% CI: 8.845-102.067) for OPN; 0.639 (95% CI: 0.538-0.729), 0.959 (95% CI: 0.909-0.982), and 41.518 (95% CI: 13.688-125.929) for AFP; and 0.856 (95% CI: 0.760-0.918), 0.738 (95% CI: 0.630-0.823), and 16.718 (95% CI: 7.950-35.156) for OPN+AFP, respectively. The area under the sROC for OPN, AFP, and OPN+AFP was 0.91, 0.88, and 0.85, respectively. For diagnosis of early HCC, pooled sensitivity of serum OPN, AFP, and OPN+AFP was 0.493 (95% CI: 0.422-0.563), 0.517 (95% CI: 0.446-0.587), and 0.732 (95% CI: 0.666-0.791), respectively. CONCLUSIONS: OPN is a comparable marker to AFP for the diagnosis of HCC, and the sensitivity of OPN was higher than that of AFP. A combination of AFP and OPN can elevate the sensitivity of diagnosis for early HCC.

Prognostic value of osteopontin in patients with hepatocellular carcinoma: A systematic review and meta-analysis.

PURPOSE: The prognostic value of tissue and serum osteopontin (OPN) in hepatocellular carcinoma (HCC) remain controversial. The aim of present meta-analysis was to evaluate the prognostic value of OPN in patients with HCC. METHODS: Eligible studies were systematically searched by PubMed, EMBASE, and Google scholar. A meta-analysis of 12 studies included 2117 cases was performed to estimate the association between OPN level and overall survival (OS), disease-free survival (DFS) in HCC patients. Subgroup analyses were also performed in the meta-analysis. RESULTS: The pooled data of studies showed that high OPN level was significantly associated with poor OS (hazard ratios [HR] 1.84; 95% confidence intervals [CI] 1.54-2.20; P = .000) and DFS (HR 1.67; 95% CI 1.40-1.98; P = .000) in HCC. Furthermore, in subgroup analysis, high tissue based OPN by immunohistochemistry detection and serum-based OPN by enzyme-linked immunosorbent assay (ELISA) detection were both significantly associated with OS (tissue: HR 1.88; 95% CI 1.53-2.31; P < .0001; serum: HR 2.38; 95% CI 1.58-3.59; P < .0001). Simultaneously, we also found that OPN expression was positively associated with stage (odds ratios [OR] 5.68; 95% CI 3.443-7.758), tumor size (Size≤5 cm vs >5 cm; OR 2.001; 95% CI1.036-3.867). CONCLUSION: The current evidence indicates that OPN could serve as a prognostic biomarker and a potential therapeutic target for HCC.

Observation and analysis of clinical efficacy of breast-conserving therapy integrated with neoadjuvant chemotherapy on Breast Cancer.

To investigate the efficaciousness of breast-conserving therapy in connection with neoadjuvant chemotherapy on breast cancer. 68 patients, who were confirmed going down with breast cancer and hospitalized from June 2015 and June 2017, were sampled and divided into two groups using the random digit table, i.e. the observation group (n=34) and the control group (n=34). Patients in the observation group experienced breast-conserving therapy integrated with neoadjuvant chemotherapy, but those in the control group received the radical resection of breast cancer. Patients' condition in surgery, incidence of post-surgery complications as well as patient survivals were compared and coded. In the observation group, surgical duration, intraoperative bleeding amount, length of stay in hospital and incidence rate of post-surgery complications were all lower than the patients with the similar conditions in the control group with evident distinctions in statistics (p<0.05). In the observation group, survival ratios of one-to-five-year living patients were evidently higher than those in the control group. The distinctions owned evident significance in calculations (p<0.05). In comparison of the recurrence ratio of disease and the rate of distant metastasis between the observation group (5.88% and 8.82%) and the control group (11.76% and 8.82%), differences had no statistical significance (p>0.05). Before treatment, compared with the score of life quality in the two groups, no evident distinction in statistical exists (p>0.05), however, after that, the life quality in the observation group evidently outweighs the quality in the control group, which shows the distinctions in statistics (p<0.05). Breast-conserving therapy in combination with neoadjuvant chemotherapy shows promising clinical value in ameliorating the life quality, decreasing the mortality rate and the incidence of adverse reaction, which is expected to be applied in clinical practices as a kind of safe and effective method.

MiR-125b inhibits anaplastic thyroid cancer cell migration and invasion by targeting PIK3CD.

Emerging evidences indicate that dysregulated microRNAs are implicated in the process of tumorigenesis and progression. The miRNA-125b (miR-125b) is downregulated and identified as tumor supressor in various cancers including thyroid cancer. However, the role and mechanism of miR-125b in anaplastic thyroid cancer (ATC) migration and invasion remain unknown. In the present study, the expression levels of miR-125b were downregulated and the expression levels of phosphoinositide 3-kinase catalytic subunit delta (PIK3CD) were upregulated in ATC tissues and cell lines. Moreover, miR-125b expression was negatively related to PIK3CD expression in ATC tissues. A computational search and luciferase assay identified PIK3CD as a direct target of miR-125b in ATC and PIK3CD expression was downregulated by miR-125b in ATC cells. In terms of function, miR-125b repressed migration and invasion of ATC cells, whereas PIK3CD overexpression reversed this effect. Furthermore, we showed that exogenous miR-125b decreased the PI3K, phospho-Akt, and phospho-mTOR expression in ATC cells. In conclusion, these results indicated that miR-125b suppressed ATC cell migration and invasion by targeting PIK3CD expression, and suggested novel potential therapeutic targets for the treatment of ATC.

Long Noncoding RNA H19 Inhibits Cell Viability, Migration, and Invasion Via Downregulation of IRS-1 in Thyroid Cancer Cells.

Thyroid cancer is a common endocrine gland malignancy which exhibited rapid increased incidence worldwide in recent decades. This study was aimed to investigate the role of long noncoding RNA H19 in thyroid cancer. Long noncoding RNA H19 was overexpressed or knockdown in thyroid cancer cells SW579 and TPC-1, and the expression of long noncoding RNA H19 was detected by real-time polymerase chain reaction. The cell viability, migration, and invasion were determined by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide assay, Transwell assay, and wound healing assay, respectively. Furthermore, cell apoptosis was analyzed by flow cytometry, and expressions of some factors that were related to phosphatidyl inositide 3-kinases/protein kinase B and nuclear factor κB signal pathway were measured by Western blotting. This study revealed that cell viability and migration/invasion of SW579 and TPC-1 were significantly decreased by long noncoding RNA H19 overexpression compared with the control group ( P < .05), whereas cell apoptosis was statistically increased ( P < .001). Meanwhile, cell viability and migration/invasion were significantly increased after long noncoding RNA H19 knockdown ( P < .05). Furthermore, long noncoding RNA H19 negatively regulated the expression of insulin receptor substrate 1 and thus effect on cell proliferation and apoptosis. Insulin receptor substrate 1 regulated the activation of phosphatidyl inositide 3-kinases/AKT and nuclear factor κB signal pathways. In conclusion, long noncoding RNA H19 could suppress cell viability, migration, and invasion via downregulation of insulin receptor substrate 1 in SW579 and TPC-1 cells. These results suggested the important role of long noncoding RNA H19 in thyroid cancer, and long noncoding RNA H19 might be a potential target of thyroid cancer treatment.

Association between single nucleotide polymorphisms of upstream transcription factor 1 (USF1) and susceptibility to papillary thyroid cancer.

BACKGROUND: Thyroid cancer, predominantly by papillary thyroid cancer (PTC), is a malignant tumour of endocrine system with increasing incidence rate worldwide. Upstream transcription factor 1 (USF1) regulates a variety of biological processes by transactivation of functional genes. In this study, we investigated the association between USF1 polymorphisms and PTC risk. MATERIAL & METHODS: A total of 334 patients with PTC, 186 patients with benign nodules (BN) and 668 healthy controls were enrolled in our study. Tag-SNPs were identified in Chinese Han in Beijing (CHB) from International HapMap Project Databases. Genomic DNAs were extracted by TaqMan Blood DNA kits. SNPs of USF1 were genotyped by TaqMan SNPs genotyping assay. Odds ratios (OR) and corresponding 95% confidence interval (CI) were used to assess the association between USF1 genetic variants and PTC risk. The statistical analyses were carried out with spss 13.0 software. RESULTS: Five tag-SNPs were retrieved to capture all the genetic variants of USF1. Among the five tag-SNPs, genetic variants in rs2516838, rs3737787 and rs2516839 have significant association with PTC risk. The rs2516838 polymorphisms dominant model (CG+GG vs CC: OR = 0·71; 95% CI: 0·52-0·97; P = 0·033) and allelic model (C vs G: OR = 0·031; 95% CI: 0·56-0·97; P = 0·031) indicated it may act as a protective factor against PTC. On the contrary, the results of rs3737787 polymorphisms: dominant model (CT+TT vs CC: OR = 1·55; 95%CI: 1·09-2·02; P = 0·001) and allelic model (C vs T: OR = 1·35; 95%CI: 1·10-1·64; P = 0·003), as well as the results of rs2516839 polymorphisms: dominant model (GA+AA vs GG: OR = 1·77; 95%CI: 1·31-2·38; P < 0·001) and allelic model (G vs A: OR = 1·36; 95%CI: 1·13-1·63; P = 0·014), revealed that they may act as risk factors for PTC. CONCLUSION: In this study, we found the SNPs of rs2516838 (mutant G alleles vs wild C alleles), rs3737787 (mutant T alleles vs wild C alleles) and rs2516839 (mutant A alleles vs wild G alleles) were significantly associated with PTC risk. Further large-scale studies with different ethnicities are still needed to validate our findings and explore the underlying mechanism of USF1 in PTC development.