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Background Breast cancer (BC) is regarded as one of the most common cancers diagnosed among the female population and has an extremely high mortality rate. It is known that Fibronectin 1 (FN1) drives the occurrence and development of a variety of cancers through metabolic reprogramming. Aspartic acid is considered to be an important substrate for nucleotide synthesis. However, the regulatory mechanism between FN1 and aspartate metabolism is currently unclear.Methods We used RNA sequencing (RNA seq) and liquid chromatography-mass spectrometry (LC-MS) to analyze the tumor tissues and paracancerous tissues of patients. MCF7 and MDA-MB-231 cells were used to explore the effects of FN1-regulated aspartic acid metabolism on cell survival, invasion, migration and tumor growth. We used PCR, Western blot, immunocytochemistry and immunofluorescence techniques to study it.Results We found that FN1 was highly expressed in tumor tissues, especially in Lumina A and TNBC subtypes, and was associated with poor prognosis. In vivo and in vitro experiments showed that silencing FN1 inhibits the activation of the YAP1/Hippo pathway by enhancing YAP1 phosphorylation, down-regulates SLC1A3-mediated aspartate uptake and utilization by tumor cells, inhibits BC cell proliferation, invasion and migration, and promotes apoptosis. In addition, inhibition of FN1 combined with the YAP1 inhibitor or SLC1A3 inhibitor can effectively inhibit tumor growth, of which inhibition of FN1 combined with the YAP1 inhibitor is more effective.Conclusion Targeting the "FN1/YAP1/SLC1A3/Aspartate metabolism" regulatory axis provides a new target for BC diagnosis and treatment. This study also revealed that intratumoral metabolic heterogeneity plays an important role in the progression of different subtypes of breast cancer.

Triple-negative breast cancer (TNBC) is extremely aggressive and lacks effective therapy. SAM and SH3 domain containing1 (SASH1) has been implicated in TNBC as a candidate tumor suppressor; however, the mechanisms of action of SASH1 in TNBC remain underexplored. Here, we show that SASH1 was significantly downregulated in TNBC patients samples compared with other subtypes of breast cancer. Ectopic SASH1 expression inhibited, while depletion of SASH1 enhanced, the invasive phenotype of TNBC cells, accompanied by deregulated expression of MMP2 and MMP9. The functional effects of SASH1 depletion were confirmed in the chicken chorioallantoic membrane and mouse xenograft models. Mechanistically, SASH1 knockdown downregulated the phosphorylation levels of the Hippo kinase LATS1 and its effector YAP (Yes associated protein), thereby upregulating YAP accumulation together with its downstream target CYR61. Consistently, forced SASH1 expression exhibited opposite effects. Pharmacological inhibition of YAP or knockdown of YAP reversed the enhanced cell invasion of TNBC cells following SASH1 depletion. Furthermore, SASH1-induced YAP signaling was LATS1-dependent, which in reverse enhanced phosphorylation of SASH1. The SASH1 S407A mutant (phosphorylation deficient) failed to rescue the altered YAP signaling by SASH1 knockdown. Notably, SASH1 depletion upregulated ARHGAP42 levels via YAP-TEAD and the YAP-ARHGAP42-actin axis contributed to SASH1-regulated TNBC cell invasion. Therefore, our findings uncover a new mechanism for the tumor-suppressive activity of SASH1 in TNBC, which may serve as a novel target for therapeutic intervention.

miR-590-3p acts as a tumor suppressor in glioblastoma multiform, medulloblastoma, hepatocellular carcinoma, and nephroblastoma. Here, we studied the role of miR-590-3p in triple-negative breast cancer (TNBC). The miR-590-3p levels in TNBC specimens were significantly lower than those in non-TNBC specimens. Overexpression of miR-590-3p significantly inhibited migration and invasion of TNBC cells and lung metastasis in vivo. Interestingly, miR-590-3p decreased the Slug mRNA and protein levels in TNBC cells, and luciferase reporter assay showed that miR-590-3p directly targeted 3’-UTR of Slug in TNBC cells. Importantly, overexpression of Slug reversed the inhibitory effect of miR-590-3p on migration and invasion of TNBC cells. Taken together, miR-590-3p inhibits TNBC migration and invasion by directly targeting Slug, suggesting a potential therapeutic effect of miR-590-3p for TNBC.

Reactivation of T cell immunity by PD-1/PD-L1 immune checkpoint blockade has been shown to be a promising cancer therapeutic strategy. However, PD-L1 immunohistochemical readout is inconsistent with patient response, which presents a clinical challenge to stratify patients. Because PD-L1 is heavily glycosylated, we developed a method to resolve this by removing the glycan moieties from cell surface antigens via enzymatic digestion, a process termed sample deglycosylation. Notably, deglycosylation significantly improves anti-PD-L1 antibody binding affinity and signal intensity, resulting in more accurate PD-L1 quantification and prediction of clinical outcome. This proposed method of PD-L1 antigen retrieval may provide a practical and timely approach to reduce false-negative patient stratification for guiding anti-PD-1/PD-L1 therapy.

Highlights • An in-house ELISA was developed to detect circulating antibodies to peptide antigens. • Circulating anti-MYC IgG levels were significantly increased in patients with NSCLC. • Neither anti-MYC nor anti-BIRC5 IgG levels significantly changed in early stage NSCLC. • Altered anti-MYC IgG levels were observed only in patients with late stage NSCLC., An in-house enzyme-linked immunosorbent assay (ELISA) was developed in this study to detect circulating IgG antibodies to peptide antigens derived from baculoviral IAP repeat-containing protein 5 isoform 2 (BIRC5) and myc proto-oncogene protein (MYC) in non-small cell lung cancer (NSCLC). Student’s t-test revealed that circulating anti-MYC IgG levels were significantly increased in patients with NSCLC compared with control subjects in the discovery sample (t = 3.96, P = 0.0001) but not in the validation sample (t = 1.24, P = 0.217), generating a combined P-value of 0.0003. Neither the discovery sample nor the validation sample showed a significant change in anti-BIRC5 IgG levels in NSCLC. Further analysis was performed to investigate whether circulating IgG antibodies to these two tumor-associated antigens (TAAs) significantly changed with early (stages I + II) and late (stages III + IV) NSCLC stages. The results showed that neither anti-MYC IgG nor anti-BIRC5 IgG levels significantly changed in patients with early stage NSCLC, while patients with late stage NSCLC had higher levels of circulating anti-MYC IgG than control subjects in the discovery sample (t = 4.74, P

MicroRNAs (miRNAs) are a class of small non-coding RNAs that, when dysregulated, are involved in the initiation and progression of various cancers, including lung cancer, in humans. In the current study, qRT-PCR was performed to measure miR-211 expression in human non-small cell lung cancer (NSCLC) cell lines and tissues. Cell proliferation, cell cycle, colony formation, and invasion were performed to detect the functional role of miR-211 in human NSCLC cell line. We used luciferase reporter assay to find the potential target of miR-211. We found that miR-211 expression was upregulated in human non-small cell lung cancer (NSCLC) cell lines and tissues. The overexpression of miR-211 enhanced NSCLC cell proliferation, colony formation, and invasion. SRC kinase signaling inhibitor 1 (SRCIN1) was identified as a direct target of miR-211. SRCIN1 silencing promoted cell proliferation, and SRCIN1 expression was downregulated in human NSCLC cell lines. Thus, miR-211 may function as an oncogenic miRNA in NSCLC, partly by regulating SRCIN1, and the modulation of miR-211 expression represents a potential strategy for the treatment of NSCLC patients.

Background: Forkhead box P3 (FOXP3) has been found to be overexpressed by a range of cancer cells and correlated with prognosis of tumors. This finding raises the possibility that the development of anti-FOXP3 antibody test may be useful for clinical application. Objective: The present work was designed to test whether circulating autoantibody to FOXP3 was altered in lung cancer. Methods: 271 patients with non-small cell lung cancer (NSCLC) and 227 in control subjects matched in age, gender and smoking history were recruited. Circulating anti-FOXP3 IgG antibody was tested using an in-house enzymelinked immunosorbent assay. Results: Student’s t-test showed that the levels of IgG autoantibody to FOXP3 were significantly higher in patients with NSCLC than control subjects (t = 7.67, P 0.65 - 0.75, SE ± 0.024), in which patients at stage 2 had the highest AUC of 0.75 (95%CI 0.67 - 0.81, SE ± 0.037), with a sensitivity of 31.4% against a specificity of 90.3%. Analysis of quality control samples gave an inter-assay deviation of 13.3% among 45 plates tested. Conclusions: Circulating IgG autoantibody to FOXP3 may be a potential biomarker for lung cancer.

Tumors can trigger specific immune response to tumor-associated antigens but the precise mechanism remains unclear. Since regulatory T-lymphocytes (Treg) play a crucial role in controlling autoimmune responses, the present work was undertaken to test whether dysfunction of Treg cells could be involved in developing autoimmunity in patients with lung cancer.In this study, we developed an in-house enzyme-linked immunosorbent assay to test circulating anti-CD25 autoantibodies among 272 patients with non-small cell lung cancer (NSCLC) and 226 control subjects matched in age, gender and smoking history.Mann-Whitney U test showed that the anti-CD25 IgG level was significantly higher in patients with NSCLC than control subjects (Z = -7.48, P 0.001) while the anti-CD25 IgA level was not significantly changed in the patient group as compared with the control group (Z = -1.34, P = 0.181). Spearman correlation analysis failed to reveal a significant correlation between the levels of anti-CD25 IgG and IgA either in patients with NSCLC (r = -0.034, P = 0.578) or in control subjects (r = 0.055, P = 0.429). ROC analysis showed an AUC of 0.70 for anti-CD25 IgG, in which NSCLC at stage III had the highest AUC (0.75). The sensitivity against a specificity of90 % was 35.0 % for anti-CD25 IgG assay with an inter-assay deviation of 9.4 %, and 4.0 % for anti-CD25 IgA assay with an inter-assay deviation of 13.0 %.Circulating anti-CD25 IgG antibody may be a useful biomarker for prognosis of lung cancer.

Overexpression of tumor-associated antigens (TAAs) has been reported in many types of cancer and may trigger secretion of their autoantibodies. The present work was thus designed to test whether circulating antibody to p16 protein-derived antigens was altered in lung cancer. Two hundred seventy-one patients with non-small cell lung cancer (NSCLC) and 226 control subjects matched in age, gender, and smoking history were recruited in this study. The levels of circulating anti-p16 IgA and IgG antibodies were tested using an enzyme-linked immunosorbent assay (ELISA) developed in-house with linear peptide antigens derived from p16 protein. Student’s t test showed that patients with NSCLC had a significant higher level of anti-p16 IgG antibody than control subjects (t = 2.74, P = 0.0063) but did not have a significant increase in IgA antibody levels (t = 1.92, P = 0.056). The sensitivity against >90 % specificity was 19.7 % for the IgG assay with an inter-assay deviation of 11.6 %, and 10.3 % for the IgA assay with an inter-assay deviation of 14.7 %. Based on a cut-off value determined by the 99th percentile of control IgG levels, the anti-p16 IgG positivity was 6.7 % in patients with NSCLC compared to 0.88 % in control subjects (χ 2 = 10.58, P = 0.001, OR = 7.97, 95 % CI 1.84–34.85). Circulating anti-p16 IgG levels were increased with stages of NSCLC, and patients with stage IV NSCLC had the highest IgG level among all four stages (t = 2.42, P = 0.016, compared with the control group). Pearson correlation analysis showed a significant correlation between circulating levels of IgA and IgG in the patient group (r = −0.2, df = 236, P = 0.0021) but not in the control group (r = −0.1, df = 205, P = 0.146). Circulating IgG antibody to p16 protein may be a potential biomarker with prognostic values for lung cancer.

One water-soluble polysaccharide (PCPw) was isolated and purified from the roots of Pulsatilla chinensis by DEAE cellulose-52 and Sephadex G-100 column chromatography, and its antitumor activity was evaluated on 4T1 tumor-bearing mice through transplantable animal tumor. After 10 days of PCPw (50, 100 and 200 mg/kg) treatment once daily in tumor-bearing mice, PCPw oral administration could not only significantly inhibit the growth of transplantable 4T1 tumor in mice but also promote concanavalin A (Con A), lipopolysaccharide (LPS)-stimulated splenocytes proliferation, the serum lysozyme level and 2,4-dinitrofluorobenzene (DNFB)-induced delayed-type hypersensitivity (DTH) reactions, especially at the dose of 100 mg/kg. Meanwhile, significant improvements in peripheral blood abnormality and anemia were observed in PCPw-treated group. These results suggested that PCPw could improve both cellular and humoral immune response and might be explored as a potential natural antitumor drug.