LncRNA MIR4435-2HG promotes proliferation, migration, invasion and epithelial mesenchymal transition via targeting miR-22-3p/TMEM9B in breast cancer.

OBJECTIVE: Breast cancer, as a malignancy with the highest incidence and mortality in women, seriously threatens women's life and health. Pieces of evidence have suggested that long non-coding RNAs (lncRNAs) possess important roles in regulating the occurrence and development of breast cancer. METHODS: RT-qPCR was used to explore the expression levels of MIR4435-2HG, miR-22-3p and TMEM9B in breast cancer tissues and cell lines. Cell viability, proliferation, migration and invasion were assessed by CCK-8 assay, Colony formation assay, Wound healing assay and Transwell assay, respectively. The effect of MIR4435-2HG on EMT progress was explored by Immunofluorescence assay and Western blot. RNA pull-down analysis and Dual-luciferase reporter assay were performed to validate the interaction between MIR4435-2HG and miR-22-3p, as well as miR-22-3p and TMEM9B. RESULTS: MIR4435-2HG was notably up-regulated in breast cancer tissues and cell lines. Additionally, down-regulation of MIR4435-2HG restrained the viability, proliferation, migration, invasion and EMT of breast cancer cells. MiR-22-3p expression was down-regulated in breast cancer tissues and cell lines, and negatively associated with MIR4435-2HG expression. Over-expression of miR-22-3p obviously inhibited the viability, proliferation, migration, invasion and EMT of breast cancer cell lines. Furthermore, TMEM9B was up-regulated in breast cancer tissues and cell lines and negatively associated with miR-22-3p expression. TMEM9B inhibition partially restored the effects of MIR4435-2HG/miR-22-3p on the viability, proliferation, migration, invasion and EMT of breast cancer cell lines. CONCLUSION: MIR4435-2HG plays a potential tumor-promoting role in the occurrence and development of breast cancer, possibly by regulating the miR-22-3p/TMEM9B axis.

Prognostic analysis of cuproptosis-related gene in triple-negative breast cancer.

Background: Cuproptosis is a copper-dependent cell death mechanism that is associated with tumor progression, prognosis, and immune response. However, the potential role of cuproptosis-related genes (CRGs) in the tumor microenvironment (TME) of triple-negative breast cancer (TNBC) remains unclear. Patients and methods: In total, 346 TNBC samples were collected from The Cancer Genome Atlas database and three Gene Expression Omnibus datasets, and were classified using R software packages. The relationships between the different subgroups and clinical pathological characteristics, immune infiltration characteristics, and mutation status of the TME were examined. Finally, a nomogram and calibration curve were constructed to predict patient survival probability to improve the clinical applicability of the CRG_score. Results: We identified two CRG clusters with immune cell infiltration characteristics highly consistent with those of the immune-inflamed and immune-desert clusters. Furthermore, we demonstrated that the gene signature can be used to evaluate tumor immune cell infiltration, clinical features, and prognostic status. Low CRG_scores were characterized by high tumor mutation burden and immune activation, good survival probability, and more immunoreactivity to CTLA4, while high CRG_scores were characterized by the activation of stromal pathways and immunosuppression. Conclusion: This study revealed the potential effects of CRGs on the TME, clinicopathological features, and prognosis of TNBC. The CRGs were closely associated with the tumor immunity of TNBC and are a potential tool for predicting patient prognosis. Our data provide new directions for the development of novel drugs in the future.

Surgical outcomes of different approaches to dissection of lymph nodes posterior to right recurrent laryngeal nerve: a retrospective comparative cohort study of endoscopic thyroidectomy via the areolar approach and via the axillo-breast approach.

Background: The American Thyroid Association (ATA) points out that lymph nodes posterior to right recurrent laryngeal nerve (LN-prRLN) should be routinely dissected. Due to the high risk of nerve injury, the lymph nodes in this area are difficult to dissect thoroughly. Although there are many approaches to endoscopic thyroidectomy, no study has been conducted on which one is more suitable. The purpose of this study was to evaluate the safety, thoroughness, related trauma, and feasibility of two widely used endoscopic thyroidectomy approaches, so as to provide a basis for the surgeon to select a better surgical approach. Methods: This retrospective study included patients who underwent ETA (n=26) and ETAB (n=36). All patients had a pathological diagnosis of papillary thyroid carcinoma (PTC) and underwent endoscopic right thyroidectomy from May 2015 to February 2022 in the Affiliated Hospital of Nantong University. The basic clinical data and surgical outcomes of the two groups were compared. Results: There was no statistical difference between the two groups in basic clinical data and oncological characteristics, which meant that the baseline data of the two groups of patients were comparable. Significant statistical significance was observed in the operation duration (149.38±44.15 vs. 119.22±45.48 min, P=0.011), drainage volume 24 h after operation (95.54±16.79 vs. 54.46±15.11 mL, P<0.001), visual analog score (VAS) 24 h after operation (3.69±1.44 vs. 2.25±1.32, P<0.001), hospitalization duration after the operation (3.19±0.75 vs. 2.25±0.44 days, P<0.001), number of lymph node dissections after right recurrent laryngeal nerve resection (0.96±1.08 vs. 2.06±1.77, P=0.007), and number of lymph node metastases after right recurrent laryngeal nerve resection (0.12±0.33 vs. 0.58±1.00, P=0.025). Besides, there was no significant difference in the numbers of central lymph node dissections and central lymph node metastases. Conclusions: Our study indicated that compared with ETA, ETAB may perform a more efficient dissection of the LN-prRLN based on less surgical trauma, which could provide a basis for the surgeon to select a better surgical approach.

LncRNA-MCM3AP-AS1 Promotes the Progression of Infantile Hemangiomas by Increasing miR-138-5p/HIF-1α Axis-Regulated Glycolysis.

Infantile hemangioma (IH) is a common benign tumor of endothelial cells in infants. Most hemangiomas are self-limited, but a few may develop and lead to serious complications that affect the normal life of children. Therefore, finding an effective treatment strategy for IH is a pressing need. Recent studies have demonstrated that non-coding RNAs affect the progression of multiple tumors. This study aims to investigate the mechanism by which LncRNA-MCM3AP-AS1 promotes glycolysis in the pathogenesis of IH. We first documented that the expression of LncRNA MCM3AP-AS1 was significantly upregulated in IH. Furthermore, we demonstrated that MCM3AP-AS1 bound to miR-106b-3p which promotes glycolysis in IH. In addition, we found that inhibition of HIF-1α contributed to the transformation of glycolysis to normal aerobic oxidation, partially reversed the promoting effect on glycolysis by the up-regulation of LncRNA MCM3AP-AS1 in IH disease. More importantly, we demonstrated this phenomenon existed in IH patients. Taken together, we demonstrate that LncRNA-MCM3AP-AS1 promotes the progression of infantile hemangiomas by increasing the glycolysis via regulating miR-138-5p/HIF-1α axis.

A novel PDGF receptor inhibitor-eluting stent attenuates in-stent neointima formation in a rabbit carotid model.

A novel drug-eluting stent (DES) is required to target vascular smooth muscle cells (SMCs) without harming endothelial cells (ECs). Platelet-derived growth factor (PDGF) is critical for the proliferation and migration of SMCs. Sunitinib [a PDGF receptor (PDGFR) tyrosine kinase inhibitor]­eluting stents may therefore inhibit neointimal formation. The aim of the present study was to examine the stent­based delivery of sunitinib in a rabbit carotid model; in addition, the effects of sunitinib were evaluated in vitro. Local administration of sunitinib markedly reduced neointimal formation without delaying re-endothelialization in the carotid artery model. In vitro, sunitinib inhibited SMC proliferation; however, no effects were observed on ECs. Sunitinib caused necrosis of SMCs. In addition, sunitinib attenuated PDGF-stimulated SMC migration in a scratch wound assay and inhibited α­SMA cytoskeleton polymerization. Furthermore, sunitinib inhibited PDGF-induced phosphorylation of extracellular signal-regulated kinase in vitro and in vivo. Therefore, this novel DES may be a potential strategy for the treatment of vascular disorders.

Triple-amiRNA VEGFRs inhibition in pancreatic cancer improves the efficacy of chemotherapy through EMT regulation.

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with dismal outcome. Both novel prognostic markers and therapeutic targets are needed to improve the overall outcome of patients. Although single or double VEGFRs have been studied in PDAC, little is known about the role of triple combination of VEGFRs (VEGFR1, 2, and 3) in prognosis and therapy. We determined VEGFRs protein expression in 241 pancreatic tissues by tissue microarray immunohistochemistry (TMA-IHC), and correlated with patients' clinical characteristics and overall survival. Subsequently, we inactivated VEGFRs expression using artificial microRNAs (amiRNAs) in vitro. Triple combination of amiRNAs to VEGFRs reduced cell proliferation, increased apoptosis, and reduced cell migration and invasion in pancreatic cancer cell lines. In the mouse xenograft pancreatic cancer model, triple VEGFRs silencing significantly reduced tumor growth, had synergistic effect with standard chemotherapy, and was associated with inhibition of epithelial mesenchymal transition (EMT). We conclude that triple combination of VEGFRs is a prognostic marker for PDAC, and inhibition of VEGFRs expression via amiRNA represents a novel targeted therapy in PDAC through regulating EMT.

Rescuing Impaired Re-endothelialization of Drug-Eluting Stents Using the Hepatocyte Growth Factor.

BACKGROUND: Current commercially available drug-eluting stents (DESs) are criticized for the problem of stent thrombosis by induced impaired re-endothelialization (RE). The solving of this challenge could be boosted by endothelial progenitor cells (EPCs). The purpose of this study was to examine the effects of hepatocyte growth factor (HGF) on this process. METHODS: The abundance and functional capacity of circulating EPC was analyzed by a fluorescence-activated cell sorter and western blot. The in vivo effect of HGF on DES patency, RE, and neointimal formation was investigated in a hypercholesterolemic rabbit model. RESULTS: After 7 days of HGF administration, the number of CD34+/CD133+ progenitor cells had increased significantly. HGF also significantly inhibited the onset of senescence of EPC due to a decrease in protein expression of p53 and p21. In the in vivo study, HGF-treated DES had a higher patency rate than the control group (11/12 vs. 6/12, P = 0.032). Moreover, the HGF-treated group exhibited better RE (control group: 69.5 ± 12.9%, HGF group: 88.8 ± 8.4%, P = 0.006), but significantly smaller areas of neointima (control group: 0.68 ± 0.15 mm2, HGF group: 0.45 ± 0.18 mm2, P = 0.02). CONCLUSION: HGF efficiently ameliorates the vascular response to stent implantation, and has an important redeeming influence on the deleterious endothelial effects of DES.

Prognostic value of LAMP3 and TP53 overexpression in benign and malignant gastrointestinal tissues.

Lysosomal associated membrane protein 3 (LAMP3) is a newly identified tumor-specific protein. It is a downstream target gene of tumor suppressor TP53 and its expression has been associated with hypoxia-induced metastasis and poor overall survival in cervical and breast cancers. However, little is known of LAMP3 protein expression in gastrointestinal cancer and its prognostic value. We determined protein expression of LAMP3 and TP53 in both gastric (n=750) and colorectal (n=479) tissues by immunohistochemistry analysis on tissue microarray (TMA), their expression was correlated with patients' clinical parameters. LAMP3 and TP53 protein expression was significantly higher in cancerous tissues compared to normal and benign tissues. In both gastric and colorectal cancers, high LAMP3 protein expression (LAMP3+) was significantly associated with tumor stage (P=0.014 and P<0.001). No correlation between LAMP3 and TP53 expression was observed. Patients with high LAMP3 expression but not high TP53 expression had a poor overall survival (for gastric cancer P<0.001, CI: 1.762-4.567; for colorectal cancer P=0.036, CI: 1.062-5.980). Our data suggest that epithelial LAMP3 expression is an independent prognostic marker for gastrointestinal cancer.

High expression of ROR2 in cancer cell correlates with unfavorable prognosis in colorectal cancer.

The receptor tyrosine kinase-like orphan receptor 2 (ROR2) is a transmembrane protein that belongs to a conserved family of tyrosine kinase receptors involved in several functional processes. ROR2 is overexpressed in various types of solid tumors; however, the expression of ROR2, as well as its functional and prognostic significance has yet to be evaluated in colorectal cancer (CRC). In this study, one-step quantitative reverse transcription-polymerase chain reaction and immunohistochemical analysis using tissue microarrays were used to evaluate ROR2 expression in CRC and to investigate the association between ROR2 expression and patient prognosis. We observed that the expression of ROR2 mRNA and protein was significantly higher in CRC specimens compared with normal, tumor-adjacent tissues (both p<0.05). Cytoplasmic ROR2 expression was related to TNM stage (p=0.041) and lymph node metastasis (N) (p=0.015). Kaplan-Meier and multivariate analyses suggested that high cytoplasmic ROR2 expression (p=0.001), poor tumor differentiation (p=0.001), and advanced TNM stage (p=0.001) and high preoperative CEA level (p<0.001) were significantly associated with unfavorable survival of CRC patients. These results suggest that ROR2 expression is correlated with malignant attributes of CRC and may serve as an indicator for poor prognosis in patients with CRC.