Chelator boosted tumor-retention and pharmacokinetic properties: development of 64Cu labeled radiopharmaceuticals targeting neurotensin receptor.

PURPOSE: Accumulating evidence suggests that neurotensin (NTS) and neurotensin receptors (NTSRs) play key roles in lung cancer progression by triggering multiple oncogenic signaling pathways. This study aims to develop Cu-labeled neurotensin receptor 1 (NTSR1)-targeting agents with the potential for both imaging and therapeutic applications. METHOD: A series of neurotensin receptor antagonists (NRAs) with variable propylamine (PA) linker length and different chelators were synthesized, including [64Cu]Cu-CB-TE2A-iPA-NRA ([64Cu]Cu-4a-c, i = 1, 2, 3), [64Cu]Cu-NOTA-2PA-NRA ([64Cu]Cu-4d), [64Cu]Cu-DOTA-2PA-NRA ([64Cu]Cu-4e, also known as [64Cu]Cu-3BP-227), and [64Cu]Cu-DOTA-VS-2PA-NRA ([64Cu]Cu-4f). The series of small animal PET/CT were conducted in H1299 lung cancer model. The expression profile of NTSR1 was also confirmed by IHC using patient tissue samples. RESULTS: For most of the compounds studied, PET/CT showed prominent tumor uptake and high tumor-to-background contrast, but the tumor retention was strongly influenced by the chelators used. For previously reported 4e, [64Cu]Cu-labeled derivative showed initial high tumor uptake accompanied by rapid tumor washout at 24 h. The newly developed [64Cu]Cu-4d and [64Cu]Cu-4f demonstrated good tumor uptake and tumor-to-background contrast at early time points, but were less promising in tumor retention. In contrast, our lead compound [64Cu]Cu-4b demonstrated 9.57 ± 1.35, 9.44 ± 2.38 and 9.72 ± 4.89%ID/g tumor uptake at 4, 24, and 48 h p.i., respectively. Moderate liver uptake (11.97 ± 3.85, 9.80 ± 3.63, and 7.72 ± 4.68%ID/g at 4, 24, and 48 h p.i.) was observed with low uptake in most other organs. The PA linker was found to have a significant effect on drug distribution. Compared to [64Cu]Cu-4b, [64Cu]Cu-4a had a lower background, including a greatly reduced liver uptake, while the tumor uptake was only moderately reduced. Meanwhile, [64Cu]Cu-4c showed increased uptake in both the tumor and the liver. The clinical relevance of NTSR1 was also demonstrated by the elevated tumor expression in patient tissue samples. CONCLUSIONS: Through the side-by-side comparison, [64Cu]Cu-4b was identified as the lead agent for further evaluation based on its high and sustained tumor uptake and moderate liver uptake. It can not only be used to efficiently detect NTSR1 expression in lung cancer (for diagnosis, patient screening, and treatment monitoring), but also has the great potential to treat NTSR-positive lesions once chelating to the beta emitter 67Cu.

Development of 18F-Labeled Vinyl Sulfone-PSMAi Conjugates as New PET Agents for Prostate Cancer Imaging.

Radiolabeled prostate-specific membrane antigen (PSMA) ligands have been rapidly adopted as part of patient care for prostate cancer. In this study, a new series of 18F-labeled PSMA-targeting agents was developed based on the high-affinity Glu-ureido-Lys scaffold and 18F-vinyl sulfones (VSs), the tumor uptake and tumor/major organ contrast of which could be tuned by pharmacokinetic linkers within the molecules. In particular, 18F-PEG3-VS-PSMAi showed the highest tumor uptake (12.1 ± 2.2%ID/g at 0.5 h p.i.) and 18F-PEG2-VS-PSMAi showed the highest tumor-to-liver ratio (T/L = 3.7 ± 1.0, 4.8 ± 1.2, and 6.3 ± 1.1 at 0.5, 1.5, and 3 h p.i. respectively). Significantly, compared with the FDA-approved 68Ga-PSMA-11, the newly developed 18F-PEG3-VS-PSMAi has an almost double tumor uptake (P < 0.0001) when tested in the same animal model. In conclusion, 18F-VS-labeled PSMA ligands are promising PET agents with prominent tumor uptake and high contrast. The lead agents 18F-PEG2-VS-PSMAi and 18F-PEG3-VS-PSMAi warrant further evaluation in prostate cancer patients.

Image-guided selection of Gd@C-dots as sensitizers to improve radiotherapy of non-small cell lung cancer.

BACKGROUND: Recently, gadolinium-intercalated carbon dots (Gd@C-dots) have demonstrated potential advantages over traditional high-Z nanoparticles (HZNPs) as radiosensitizers due to their high stability, minimal metal leakage, and remarkable efficacy. RESULTS: In this work, two Gd@C-dots formulations were fabricated which bore carboxylic acid (CA-Gd@C-dots) or amino group (pPD-Gd@C-dots), respectively, on the carbon shell. While it is critical to develop innovative nanomateirals for cancer therapy, determining their tumor accumulation and retention is equally important. Therefore, in vivo positron emission tomography (PET) was performed, which found that 64Cu-labeled pPD-Gd@C-dots demonstrated significantly improved tumor retention (up to 48 h post injection) compared with CA-Gd@C-dots. Indeed, cell uptake of 64Cu-pPD-Gd@C-dots reached close to 60% of total dose compared with ~ 5% of 64Cu-CA-Gd@C-dots. pPD-Gd@C-dots was therefore further evaluated as a new radiosensitizer for non-small cell lung cancer treatment. While single dose radiation plus intratumorally injected pPD-Gd@C-dots did lead to improved tumor suppression, the inhibition effect was further improved with two doses of radiation. The persistent retention of pPD-Gd@C-dots in tumor region eliminates the need of reinjecting radiosensitizer for the second radiation. CONCLUSIONS: PET offers a simple and straightforward way to study nanoparticle retention in vivo, and the selected pPD-Gd@C-dots hold great potential as an effective radiosensitizer.

Analysis of presacral tissue structure in LARS and the prevention of LARS by reconstruction of presacral mesorectum with pedicled greater omentum flap graft.

BACKGROUND: The greater omentum has played a unique biological role in regenerative surgery. The aim of our study was to alter the anterior sacral structure by filling the anterior sacral space with the greater omentum and evaluate its effect on the low anterior resection syndrome (LARS) after total mesorectal excision (TME) surgery for low rectal cancer. METHODS: We retrospectively collected clinical data of patients with primary low rectal cancer who underwent TME and ileostomy closure in our hospital from March 2018 to March 2020. Spearman correlation analysis was conducted to analyze the correlation between postoperative mesorectal fascia (MRF) thickness and LARS score. Subsequently, we prospectively used a tipped greater omental flap graft to reconstruct the anterior rectal sacral structures (MRF reconstruction) in 17 patients and compared LARS scores and rectal compliance (RC) at week 12 after closure of the ileostomy in both groups. RESULTS: There were 47 patients with No-MRF reconstruction (31 males, mean age 60.68 ± 9.21 years) and 17 with MRF reconstruction (10 males, mean age 49.82 ± 14.74 years). Correlation analysis indicated that MRF thickness and RC were negatively correlated with LARS severity (p < 0.05). The LARS score of patients with MRF reconstruction at 12 weeks was significantly better than that of those with No-MRF reconstruction (32.97 ± 2.65 vs. 26.94 ± 1.52, p = 0.001), and the RC of MRF reconstruction were lower (2.80 ± 0.55 vs. 3.67 ± 0.38, p = 0.001). In addition, MRF reconstruction and No-MRF reconstruction have the similar incidence of postoperative complications (p = 0.156). No hemorrhage or necrosis of the greater omentum flap was observed in any of the patients. CONCLUSIONS: Greater omentum flap transplantation can significantly improve the symptoms of LARS at 12 weeks after ileostomy closure and we expect it to become a new surgical procedure for the treatment of low rectal cancer.

Synthesis, Structure, and Electrochemical Properties of Some Cobalt Oxalates.

Transition-metal oxalates have wide applications in magnetics, photoemission, electrochemistry, etc. Herein, using hydrothermal reactions, five cobalt(II) oxalates, Na2Co2(C2O4)3·2H2O (I), Na2Co(C2O4)2·8H2O (II), KLi3Co(C2O4)3 (III), Li4Co(C2O4)3 (IV), and (NH4)2Co2(C2O4)F4 (V) have been synthesized, and their structures are determined from single-crystal X-ray diffraction or Rietveld refinement of powder X-ray diffraction data. Notably, IV and V are identified for the first time. The structures of these cobalt oxalates are versatile, covering 0D, 1D, 2D, and 3D frameworks, while the coordination environments of Co2+ centers are uniquely distorted octahedra. As representative examples, I and III are investigated as cathode materials for secondary batteries. Both exhibited electrochemical activity despite large cell polarization. The present study enriches the transition-metal oxalate family and provides new options for energy storage materials.

Oral squamous cell carcinoma-derived exosomes promote M2 subtype macrophage polarization mediated by exosome-enclosed miR-29a-3p.

This study aims to explore the mechanism of the signal transmission between oral squamous cell carcinoma (OSCC) and unpolarized stromal immune macrophages mediated by OSCC-derived exosomes (OSCC-Exo). Polarization of macrophages was found by detection of the level of protein markers or specific components for M1 subtype or M2 subtype macrophages, respectively. Exosomes extracted from two OSCC cell lines, which might have been transfected with micro-RNA (miR)-29a-3p inhibitor or mimic, were cocultured with macrophages to ensure the effect of exosome-enclosed miR-29a-3p on the polarization of macrophages. miR-29a-3p is highly expressed, suppressor of cytokine signaling 1 (SOCS1) is low expressed and phosphorylated signal transduction and transcriptional activator 6 (p-STAT6) is highly expressed in OSCC tissues. Upregulation of miR-29a-3p is observed in OSCC-derived exosomes. When cocultured, OSCC-derived exosomes promote M2 subtype macrophage polarization and the medium of the coculture promotes the proliferation and invasion of SCC-9 and CAL-27 cells. After interfered silencing miR-29a-3p of OSCCs, SCC-9- and CAL-27 cell-derived exosomes inhibit M2 subtype macrophage polarization. On the other hand, cellular highly expressed miR-29a-3p of macrophages enhances M2 subtype macrophage polarization. Moreover, such macrophages promote the proliferation and invasion of SCC-9 and CAL-27. SOCS1 is a direct target for miR-29a-3p and could be negatively regulated by miR-29a-3p. Moreover, SOCS1 overexpression reverses the activity of SOCS1/STAT6 signals of macrophages and cell proliferation and invasion of OSCCs induced by miR-29a-3p overexpression. Also, overexpressed SOCS1 in macrophages counteracts the impact of OSCC-derived exosomes in M2 subtype macrophage polarization. Exosome-enclosed miR-29a-3p promotes tumor growth in nude mice with xenograft. OSCC-derived exosomes promote M2 subtype macrophage polarization mediated by exosome-enclosed miR-29a-3p, and the mechanism by miR-29a-3p is the activity of SOCS1/STAT6 signals in macrophages.

Reconstruction of Maxillary and Orbital Floor Defect With Free Fibula Flap and Whole Individualized Titanium Mesh Assisted by Computer Techniques.

PURPOSE: We sought to investigate the clinical application of free fibula flap and individualized titanium mesh through the use of a virtual planning and guiding template to assist the reconstruction of maxilla and orbital floor defects. PATIENTS AND METHODS: Between 2015 and 2016, a total of 6 adult patients with maxillary and orbital floor defects were enrolled in this study. Preoperative virtual planning, including virtual maxillary resection and fibular reconstruction, was performed in all cases according to 3-dimensional radiographic and clinical findings. A 3-dimensionally printed resin model and prebent templates were used to guide the harvesting and positioning of the fibula flap during surgery. Then, an individualized titanium mesh was used to support the orbital floor and restore the maxillary contour. The results were confirmed by postoperative computed tomography scans and clinical follow-up. RESULTS: Preoperative virtual planning and prebent templates can be used to guide the harvesting and positioning of the fibula flap, as well as the forming and positioning of the individualized titanium mesh, with satisfactory results. All flaps survived, and symmetrical facial contours were achieved with normal lower jaw movement and proper vertical distance for dental implants in all patients. CONCLUSIONS: Computer-aided techniques such as virtual planning, 3-dimensionally printed models, and prebent guide templates can be used to harvest and position a free fibula flap, form personalized titanium mesh, and ultimately improve the clinical efficacy of maxillary and orbital floor reconstruction.

A retrospective study on application of fibula/iliac flap surgical techniques to mandibular defects.

This study group consists of a total of 61 patients who underwent fibula flap and iliac flap surgeries to repair mandibular defects. Patients' Quality Of life (QOL) at 6 and 24 months after surgery is investigated and compared by the EORTC-QLQ-H&N and OHIP-14. The base data of the two groups of patients are collected and analysed by the SPSS 20.0 statistical software. Independent sample t test was conducted for EORTC-QLQ-H&N and OHIP-14 scores at two time points in each group. The 61 cases of free flap all survived and the difference in the location of the primary tumor between the two groups is statistically significant. The EORTC-QLQ-H&N showed that the score of speech, diet, social contact, and teeth all went up at 6 months after surgery, but went down dramatically at 24 months after surgery. The OHIP-14 showed that there was significant reduction in functional limitation at 24 months after surgery, with statistical significance (p < 0.05) between the groups of iliac flap (19.16 ± 5.33) and fibula flap (33.77 ± 7.71). Therefore, it is suggested that patients suffering from mandibular defects receive surgery utilizing the iliac flap, while those with a larger range of defects or lesions involving the condyle and chin should receive corrective surgery utilizing the fibular flap.

Effect of Esophagus-Remnant Stomach Anterior Wall Anastomosis on Postoperative Reflux and Nutritional Status in Patients with Early- and Intermediate-Stage Proximal Gastric Cancer.

Purpose: To compare and analyze the incidence of postoperative reflux esophagitis (RE) in patients with early- and intermediate-stage proximal gastric cancer after proximal gastrectomy plus esophagus-remnant stomach anterior wall anastomosis with proper spacing between the reserved anastomotic stoma and the stump of the remnant stomach versus total gastrectomy plus Roux-en-Y anastomosis and to analyze the advantages and disadvantages of these anastomosis approaches. Methods: Hospitalization data of 23 patients with early- and intermediate-stage proximal gastric cancer were retrospectively analyzed. They were divided into an observation group who underwent proximal gastrectomy plus esophagus-remnant stomach anterior wall anastomosis with proper spacing between the reserved anastomotic stoma and the stump of the remnant stomach and a control group who underwent total gastrectomy plus Roux-en-Y anastomosis. Quality observation indicators were compared between the two groups. Results: There was no statistically significant difference between the groups in the number of lymph nodes cleared or the recurrence rate at 12 months postoperatively. The incidence of postoperative RE was significantly lower in the observation group (25%) than in the control group (80%). The operation time, postoperative length of hospital stay, appetite change, body mass index, and hemoglobin level at 6 months postoperatively were significantly better in the observation group than in the control group (P < .05). Conclusion: Proximal gastrectomy plus esophagus-remnant stomach anterior wall anastomosis with proper spacing between the reserved anastomotic stoma and the stump of the remnant stomach can be used as a preferred surgical procedure for early- and mid-stage proximal gastric cancer.

Oncology Outcomes of Total Mesorectal Excision with Preservation of Both the Superior Rectum Artery and Left Colonic Artery for Upper-Rectal and Sigmoid Colon Cancers: A Meta-Analysis.

Background: Total mesorectal resection (TME) has become the standard surgical procedure for resection of colorectal cancer tumors. We presented a systematic meta-analysis to evaluate the surgical outcomes of laparoscopic TME surgery with preservation or nonpreservation of both the superior rectum artery (SRA) and left colonic artery (LCA) for upper-rectal and sigmoid colon cancers. Methods: The comparative studies were systematically searched on PubMed, Science Direct, Web of Science, Wanfang Data, and China National Knowledge Infrastructure (CNKI) up to April 2021. Primary outcomes were oncology outcomes. And secondary outcomes involved surgical outcomes of interest and postoperative recovery. Results: Five relevant studies with a total of 761 patients undergoing laparoscopic TME surgery were eligible for meta-analysis. Three hundred seven patients received TME with preservation of both SRA and LCA (Group A), and 454 received TME surgery alone (Group B), respectively. Our results indicated that Group A had a less total postoperative complications (P = .000), lower anastomotic leakage rate (P = .002), shorter length of stay (P = .008), and longer operative time (P = .002). However, there was no significant difference between the two groups in terms of lymph node dissections (P = .188), intraoperative bleeding (P = .474), the first postoperative defecation (P = .943), recurrence rate (P = .547), and conversive rate (P = .504). Conclusions: Based on our meta-analysis, laparoscopic TME surgery with preservation of both the SRA and LCA for upper-rectal and sigmoid colon cancers may significantly receive better clinical and surgical outcomes. More well-designed large sample studies are required to replicate the short-term benefits and long-term oncologic outcomes.

The role of folded fibular flap in patients' reconstruction of mandibular defects: a retrospective clinical study.

This study has analyzed 41 patients with mandibular ameloblastoma who underwent a partial mandibulectomy and reconstruction by folding the free fibular flap. In the preoperative and postoperative (6 months and 24 months after surgery), the Quality of Life (QOL) of these patients was assessed by using the University of Washington Quality of Life Questionnaire (UW-QOL) and the medical outcome study short form-36 (SF-36) questionnaires. SPSS 20.0 statistical software was used to conduct statistical analysis on the base data of the two groups of patients. Independent sample t test was conducted for sf-36 and UW-QOL scores at two time points in each group. The SF-36 survey showed that body pain (54.54 ± 8.10), general health (55.27 ± 7.54), and health changes (58.29 ± 9.60) decreased significantly at 6 months after surgery, but the mean score at 24 months after surgery all exceeded the preoperational level. At 24 months after the surgery, the vitality (80.41 ± 3.74), social function (81.61 ± 4.07), emotional role (82.39 ± 4.07), psychological health (81.66 ± 4.37) and total score (704.00 ± 31.53) all returned to the preoperative level, which was statistically significant compared with 6 months after surgery. However, there was no significant difference compared with the preoperative level. The UW-QOL survey showed that chewing (56.68 ± 7.23), speech (54.54 ± 7.7) and taste (62.29 ± 10.15) have significantly changed at 6 months after the surgery, and the difference was statistically significant at 24 months after surgery. Saliva generation decreased slightly (80.76 ± 3.35) at 6 months after surgery, but quickly returned to the preoperative level (81.59 ± 4.06). The total score of the patients almost recovered to the preoperative level at 24 months after surgery. The folded the fibular flap can not only repair the defects of soft tissue and bone tissue, but also restore the height of the alveolar ridge to, avoid the imbalance of crown and root ratio after implantation and reduce the occurrence of peri-implant inflammation, so that a true functional reconstruction can be realized.

Upregulation of ADAM12 Is Associated With a Poor Survival and Immune Cell Infiltration in Colon Adenocarcinoma.

BACKGROUND: A disintegrin and metalloprotease 12 (ADAM12) is a member of the multidomain protein family, but the mechanisms by which it affects prognosis and immune cell infiltration in patients with colon adenocarcinoma (COAD) remain unclear. Here, our study aimed to analyze the prognostic value of ADAM12 and investigate the correlation between ADAM12 expression and immune cell infiltration in patients with COAD. METHODS: Differential expression analyses were performed using the Oncomine and UALCAN databases, and prognostic analyses were conducted using PrognoScan, Gene Expression Profiling Interactive Analysis (GEPIA), and Kaplan-Meier Plotter. Then, the cBioPortal database was used to analyze alterations in the ADAM12 gene, and the STRING and Metascape websites were used to conduct Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. Additionally, relationships between ADAM12 and the immune microenvironment were evaluated based on the TIMER, GEPIA, and TISIDB databases. RESULTS: ADAM12 was overexpressed in COAD tissues, and higher ADAM12 expression correlated with a worse prognosis for patients with COAD. The gene regulatory network suggested that ADAM12 was mainly enriched in extracellular matrix (ECM) organization, ECM proteoglycans, skeletal system development, and ossification, among other pathways. Moreover, ADAM12 expression significantly correlated with the abundance of CD4+ T cells, B cells, CD8+ T cells, neutrophils, macrophages, dendritic cells, and their markers, as well as lymphocytes, immunomodulators, and chemokines. CONCLUSIONS: In colorectal tumors, ADAM12 may play vital roles in regulating the ECM and the recruitment of immune cells, and we suggest that ADAM12 will become a reliable biomarker for determining response to immunotherapy and the prognosis of patients with COAD.

Retraction Note: MicroRNA-27a-3p Modulates the Wnt/ß-Catenin Signaling Pathway to Promote Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Stem Cells by Targeting SFRP1.

Editor's Note: this Article has been retracted; the Retraction Note is available at https://www.nature.com/articles/s41598-020-77203-x.

[Clinic analysis of 198 elder patients with maxillofacial fracture].

OBJECTIVE: The purpose of the study was to analyze the characteristics of elder patients with maxillofacial fracture. METHODS: We retrospectively analyzed the characteristics of maxillofacial fractures in the elder patients, who were treated from July 2010 to October 2017. The clinical characteristics of the etiology, fracture site, combined injury, systemic disease, and treatment method were analyzed. RESULTS: In the 198 elderly patients with maxillofacial fractures, the male-to-female ratio was 3.95︰1, and the mean age was 66.15 years old. Traffic accident injury (78 patients, 39.39%), fall injury (49 patients, 24.75%), high fall injury (33 patients, 16.67%) were the main factors of maxillofacial fracture in elderly patients. The most frequently observed fracture site was the mandible (120 patients). A total of 60 patients demonstrated associated injuries, in which limb injuries were the most prevalent (28 patients); whereas 66 patients had other systemic medical conditions, in which cardiovascular diseases was the most frequent (50 patients). The main treatment method of 198 patients was rigid internal fixation with small or micro-plates. CONCLUSIONS: Falling and traffic accidents are the main factors of maxillofacial fracture in elderly patients. Thus, interference measures should be observed for the prevention of maxillofacial fractures in elderly patients.

MicroRNA-542-3p inhibits oral squamous cell carcinoma progression by inhibiting ILK/TGF-ß1/Smad2/3 signaling.

In this study, we investigated the effects of microRNA-542-3p (miR-542-3p) on ILK/TGF-ß1/Smad2/3 signaling and oral squamous cell carcinoma (OSCC) progression. Levels of miR-542-3p were lower in OSCC tissues (n=108) than adjacent normal tissues, whereas levels of ILK, TGF-ß1 and Smad2/3 were higher. Patients with undifferentiated tumors, advanced TNM stage and lymph node metastasis showed low miR-542-3p levels. This was accompanied by high ILK expression and poor survival. Dual luciferase reporter assays of SCC-9 cells showed that miR-542-3p inhibited ILK gene expression by binding to its 3'UTR at 233-240 bp. SCC-9 cells transfected with miR-542-3p mimics exhibited elevated miR-542-3p and decreased ILK, TGF-ß1 and Smad2/3 expression. They also showed reduced self-renewal (fewer CD44+ cells and tumor-spheres), invasiveness, migration, proliferation and survival. Conversely, miR-542-3p inhibitors promoted increased self-renewal (more CD44+ cells and tumor-spheres), invasiveness, migration, proliferation and survival. In xenograft experiments with nude mice, SCC-9 cells transfected with miR-542-3p mimics or siRNA-ILK yielded tumors with smaller volumes and weights than control tumors. These results demonstrate that miR-542-3p is a tumor suppressor that inhibits ILK/TGF-ß1/Smad2/3 signaling, thereby inhibiting OSCC progression.

Induction of oxidative stress and cell apoptosis by selenium: the cure against oral carcinoma.

Oral carcinoma (OC) remains one of the most difficult malignancies to cure. selenium (Se) is an essential trace mineral for human and animals, but high concentrations of Se induce apoptosis and oxidative effects. Although cell apoptosis has been evidenced as a critical mechanism mediating the anticancer activity of Se, the underlying molecular mechanisms remain elusive. To explore the role of Se in rat OC, we examined the weather the oxidative stress-mediated apoptotic pathway induced by Se was involved in the development of OC. In this study, we successfully constructed the OC rat model by 4-Nitroquinoline-1-oxide (4-NQO) exposure which reflected from histopathological observations. Se-induced the productions of methane dicarboxylic aldehyde (MDA) and reactive oxygen species (ROS), which was accompanied by the inhibition of superoxide dismutase (SOD) both in vivo and vitro. The anti-apoptotic gene (Bcl-2) was down-regulated and pro-apoptosis members (Bax, Bak, Cyt-c, caspase9 and caspase3) were up-regulated by Se in OC cells. Meanwhile, we also found that Se could strongly inhibited the cell proliferation of OC lines in vitro. These results suggested that excessive Se could effectively cause oxidative stress and induce apoptosis in OC cells, as a result the OC was also inhibited to some extent. Therefore, the information presented in this study is believed to be helpful in supplementing data for further therapy of OC.

MicroRNA-27a-3p Modulates the Wnt/ß-Catenin Signaling Pathway to Promote Epithelial-Mesenchymal Transition in Oral Squamous Carcinoma Stem Cells by Targeting SFRP1.

This study aimed to elucidate how microRNA27a-3p (miR-27a-3p) modulates the Wnt/ß-catenin signaling pathway to promote the epithelial-mesenchymal transition (EMT) in oral squamous carcinoma stem cells (OSCSCs) by targeting secreted frizzled-related protein 1 (SFRP1). Flow cytometry was used to sort OSCSCs from the SCC-9 and Tca8113 cell lines. The OSCSCs were randomly assigned into the miR-27a-3p inhibitors group, the miR-27a-3p inhibitors-NC group, the si-SFRP1 group, the si-SFRP1 + miR-27a-3p inhibitors group and the blank group. A luciferase reporter, immunofluorescence and Transwell assays were performed to detect luciferase activity, SFRP1, and cell migration and invasion, respectively. The mRNA expression of miR-27a-3p, SFRP1 and EMT markers (E-cadherin, N-cadherin, vimentin and ZEB1) were detected using qRT-PCR. The protein expression of SFRP1, EMT markers and the proteins of the Wnt/ß-catenin signaling pathway was detected by Western blotting. OSCSCs showed up-regulated miR-27a-3p, Wnt/ß-catenin signaling pathway-related proteins, vimentin, N-cadherin and ZEB1 and down-regulated SFRP1 and E-cadherin. MiR-27a-3p targeted SFRP1. Down-regulated miR-27a-3p resulted in increased E-cadherin and SFRP1 but decreased vimentin, N-cadherin, ZEB1, the Wnt/ß-catenin signaling pathway-related proteins, and invasive and migratory cells. Silenced SFRP1 reversed this effect. We found that miR-27a-3p modulated the Wnt/ß-catenin signaling pathway to promote EMT in OSCSCs by down-regulating SFRP1.

Regulation of tumorigenesis in oral epithelial cells by defined reprogramming factors Oct4 and Sox2.

Oct4 and Sox2 are pluripotent stem cell factors but the interplay between them in tumorigenesis is unclear. The aim of the present study was to investigate the roles of Oct4 and Sox2 in the reprogramming of oral cancer stem cells. One or both Oct4 and Sox2 were overexpressed in immortalized oral epithelial (hTERT+-OME) cells by lentivirus transduction. In addition, Oct4 and Sox2 proteins in two oral squamous cell carcinoma cell (OSCC) lines (Cal27 and primary cultured OSCC from a T2N2M0 patient) were individually or combinedly knocked down by shRNA. The results showed that the doubly transduced (Oct4+Sox2+) cells could trigger neoplasms in immunodeficient mice after lentivirus transduction, but single transduced (Oct4+ or Sox2+) cells had no tumor formation ability. The knockdown Sox2low and knockdown Oct4lowSox2low cells resulted in decreased tumor size in the immunodeficient mice but the single knockdown Oct4low cancer cells acquired more aggressive xenografts. Our findings suggest that Oct4+Sox2+ cells may be reprogrammed cancer stem cells inducing oral carcinogenesis.

Polypeptide Fraction from Arca subcrenata Induces Apoptosis and G2/M Phase Arrest in HeLa Cells via ROS-Mediated MAPKs Pathways.

Arca subcrenata is documented in the literature of marine Traditional Chinese Medicine. Polypeptide fraction from A. subcrenata, coded as P2, was demonstrated to possess significant antitumor activity in our previous study. However, the underlying mechanism remains undefined. The present study was carried out to investigate the underlying antitumor mechanism of P2 in human cervical cancer HeLa cells by MTT, FCM, LSCM, and western blot assays. The results revealed that P2 significantly induced apoptosis of HeLa cells in a concentration- and time-dependent manner. High level of ROS was provoked by P2, which was in turn responsible for induction of apoptosis through activation of intrinsic mitochondrial pathway and JNK1/2, p38 MAPK pathways, as well as inhibition of ERK1/2 pathway, as evidenced by the abrogation of P2's effect on HeLa cells preincubated with the ROS scavenger NAC. P2 also was observed to display significant effect on G2/M phase arrest by downregulating the expression of cyclin B1/cdc2 complex and upregulating the expression of p21. These findings demonstrate that P2 induces apoptosis and G2/M phase arrest in HeLa cells through ROS-mediated MAPKs pathways, suggesting that P2 would be worth investigating as a promising agent within the scope of marine drugs for treatment of cervical cancer.