LINC01133 regulates MARCKS expression via sponging miR-30d-5p to promote the development of lung squamous cell carcinoma.

LncRNAs are vital regulators for lung squamous cell carcinoma (LUSC). However, the detailed role that LINC01133 plays in LUSC is unclear. This work sought to explore the potential function of LINC01133.Levels of LINC01133, miR-30d-5p, and MARCKS were separately tested in both tissues and cells using qRT-PCR. Proliferation was assessed through MTT experiment and apoptosis was detected upon flow cytometry. Transwell experiments were implemented to evaluate migratory and invasive abilities. The interaction between two genes was affirmed through luciferase reporter assay and RNA pull-down experiment. Western blotting measured the protein level of MARCKS. Animal models were established and tissues were taken for IHC analysis of MARCKS and Ki67.LINC01133 was elevated in LUSC and its downregulation could suppress proliferation, migration and invasion but induced apoptosis. LINC01133 interacted with and regulated the binding of miR-30d-5p to MARCKS. LINC01133/miR-30d-5p axis mediated proliferation, apoptosis, migration and invasion in LUSC cells, as well as modulated tumor growth in animal models. LINC01133 interacted with miR-30d-5p to modulate MARCKS expression, contributes to promoted cell proliferation, migration, invasion, and inhibited cell apoptosis in vitro, and promoted tumor growth in vivo. These findings could provide possible therapeutic targets in view of LUSC treatment in the future.

A Modular and Cost-Effective Droplet Microfluidic Device for Controlled Emulsion Production.

The droplet microfluidic device has become a widely used tool in fields such as physics, chemistry, and biology, but its complexity has limited its widespread application. This report introduces a modular and cost-effective droplet microfluidic device for the controlled production of complex emulsions, including oil and aqueous single emulsions, and double emulsions with varying numbers of encapsulated droplets. The droplet sizes can be precisely controlled by easily replacing flat needles and adjusting the needle position within an axially accelerated co-flow field. This modular device not only allows for easy repair and maintenance in case of device clogging or damage but can also be readily expanded to produce complex emulsions. The low-cost and user-friendly nature of the device greatly facilitates the widespread adoption and utilization of droplet microfluidics.

[Retracted] MACC­1 antibody target therapy suppresses growth and migration of non­small cell lung cancer.

Following the publication of this paper, it was drawn to the Editor's attention by a concerned reader that the control data in Fig. 2B and C on p. 7332, showing immunofluorescence and migration assay experiments respectively, were strikingly similar to data appearing in different form in another article which was written by different authors at different research institutes [Tian L, Shen D, Li X, Shan X, Wang X, Yan Q and Liu J: Ginsenoside Rg3 inhibits epithelial­mesenchymal transition (EMT) and invasion of lung cancer by down­regulating FUT4. Oncotarget 7: 1619­1632, 2016]. Owing to the fact that the contentious data in the above article had already been published prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 16: 7329­7336, 2017; DOI: 10.3892/mmr.2017.7517].

Over-expression of USP15/MMP3 predict poor prognosis and promote growth, migration in non-small cell lung cancer cells.

Aberrant ubiquitin modifications caused by an imbalance in the activities of ubiquitinases and de-ubiquitinases are emerging as important mechanisms underlying non-small cell lung cancer (NSCLC) progression. The deubiquitinating enzyme ubiquitin-specific peptidase 15 (USP15) has been identified as an important factor in oncogenesis and a potential therapeutic target. However, the expression profile and function of USP15 in NSCLC remain elusive. In the present study, we investigated the expression pattern and the potential biological functions of USP15 in NSCLC both in cells and animal models. Our data revealed that USP15 was highly expressed in NSCLC tissues and cells compared with normal counterpart. We subsequently knocked down USP15 expression in two NSCLC cell lines, which significantly suppressed cell proliferation. In addition, knocking down USP15 expression reduced NSCLC cell migration and invasion according to the results from Matrigel-Transwell analysis. NSCLC animal model results showed that USP15 knockdown also reduced NSCLC size. Biochemical analysis revealed that USP15 knockdown inhibited matrix metalloproteinase (MMP)3 and MMP9 expression. Furthermore, high levels of USP15 and MMP3 expression were associated with poor prognosis in NSCLC. In conclusion, the results from the present study suggest that the high expression of USP15 promotes NSCLC tumorigenesis. Therefore, it is proposed that USP15 and MMPs may represent novel biomarkers for NSCLC progression and prognosis.

Self-assembly of photosensitive and radiotherapeutic peptide for combined photodynamic-radio cancer therapy with intracellular delivery of miRNA-139-5p.

BACKGROUND: A significant challenge in cancer therapy is to maximize the therapeutic efficacy and minimize the side effects. In the past decade, a lot of nanoparticles have been used as the carriers for efficient drug delivery. METHODS AND RESULTS: This study was to prepare R9 modified with 125I-labeled cRGD and ce6 which self-assembled with miR-139-5p to form nanoparticles (Ce6-R9-125I-RGD-MNPs), and to further take advantage of the enhanced permeability and retention (EPR) effect of radiolabeled nanoparticles to realize the integration of tumor diagnosis and treatment. We successfully synthesized and represented it, saline and serum stability experiments demonstrating good stability. Moreover, Ce6-R9-125I-RGD-MNPs showed superior tumor targeting and the effect of combined photodynamic therapy (PDT) and radiotherapy treatment in vivo and vitro. CONCLUSION: The pathological results further confirmed that the therapeutic doses of Ce6-R9-125I-RGD-MNPs cause pathological changes of tumor tissues while showing minimal toxicity to normal tissues.

Downregulation of circRNA_100876 Inhibited Progression of NSCLC In Vitro via Targeting miR-636.

BACKGROUND: Non-small cell lung carcinoma (NSCLC) is a common malignant tumor with poor prognosis. CircRNA-100876 has been considered to be involved in NSCLC. However, the mechanism by which circRNA_100876 mediated the progression of NSCLC remains unclear. METHODS: CCK8 assay and immunofluorescence were used to detect cell proliferation. Flow cytometry and transwell assay were performed to analyze cell apoptosis, migration and invasion, respectively. Verification of possible target for circRNA_100876 and related miR-636 were done using luciferase assay. In addition, western blot was performed to detect the protein expressions in NSCLC cells. RESULTS: Silencing of circRNA_100876 notably inhibited the proliferation of NSCLC cells. Moreover, downregulation of circRNA_100876 significantly induce the apoptosis of NSCLC cells via mediation of apoptosis-related proteins. In addition, silencing of circRNA_100876 significantly inhibited migration and invasion of NSCLC cells. MiR-636 was the downstream target of circRNA_100876. Meanwhile, RET was the direct target of miR-636. Finally, circRNA_100876 shRNA2 notably suppressed the progression of NSCLC through PI3K/Akt signaling. CONCLUSION: CircRNA_100876 knockdown notably suppressed the progression of NSCLC through regulation of miR-636/RET axis, which may serve as a potential target for treatment of NSCLC.

Downregulation of miR-7-5p Inhibits the Tumorigenesis of Esophagus Cancer via Targeting KLF4.

BACKGROUND: Esophageal cancer (EC) is one of the aggressive gastrointestinal malignancies. It has been reported that microRNAs (miRNAs) play key roles during the tumorigenesis of EC. To identify novel potential targets for EC, differential expressed miRNAs (DEG) between EC and adjacent normal tissues were analyzed with bioinformatics tool. METHODS: The differential expression of miRNAs between EC and adjacent normal tissues was analyzed. CCK-8 and Ki67 staining were used to detect the cell proliferation. Flow cytometry was performed to test the cell apoptosis. The correlation between miR-7-5p and KLF4 was detected by dual-luciferase report assay. Gene and protein expression in EC cells or in tissues were measured by qRT-PCR and Western blot, respectively. Cell migration and invasion were detected with transwell assay. Xenograft mice model was established to investigate the role of miR-7-5p in EC tumorigenesis in vivo. RESULTS: MiR-7-5p was found to be negatively correlated with the survival rate of patient with EC. In addition, downregulation of miR-7-5p significantly inhibited the growth and invasion of EC cells. Meanwhile, miR-7-5p directly targeted KLF4 in EC cells. Moreover, downregulation of miR-7-5p inhibited the tumorigenesis of EC via inactivating MAPK signaling pathway in vivo. CONCLUSION: Downregulation of miR-7-5p notably suppressed the progression of EC via targeting KLF4. Thus, miR-7-5p might serve as a new target for the treatment of EC.

Silence of FOXD2-AS1 inhibited the proliferation and invasion of esophagus cells by regulating miR-145-5p/CDK6 axis.

This study aimed to investigate the function of long non-coding RNA FOXD2 adjacent opposite strand RNA 1 (lncRNA FOXD2-AS1) during the progression of esophagus cancer (EC) and explore its underlying molecular mechanisms. The level of FOXD2-AS1 in EC tissues and paracancerous tissues was detected by using RT-qPCR; ROC curve was used to evaluate the diagnostic value of FOXD2-AS1 for EC. In addition, CCK8 assay and immunofluorescence staining assay were used to detect the proliferation of Eca-109 and TE-1 cells. To investigate the function of FOXD2-AS1 on cell apoptosis and cell cycle, flow cytometry was performed. To detect the invasion ability of EC cells, transwell invasion assay was performed. Starbase3.0 and Targetscan were used to predict the target genes of FOXD2-AS1 and miR-145-5p, and protein expressions were detected with western blot. We found FOXD2-AS1 was significantly upregulated in EC tissues compared with adjacent normal tissues, which was positively correlated with clinicopathological parameters of patients with EC. Downregulation of FOXD2-AS1 inhibited the proliferation and invasion by inducing apoptosis of EC cells. Moreover, FOXD2-AS1 may regulate the expression of CDK6 by targeting miR-145-3p. Meanwhile, silencing of FOXD2-AS1 caused G1 phase arrest of EC cells by reducing the expression of CDK6. In conclusion, silening FOXD2-AS1 significantly inhibited the proliferation and invasion of EC cells by regulating the miR-145-5p/CDK6 axis. Therefore, FOXD2-AS1 might be used as diagnostic biomarker and therapeutic target for EC.

Grim-19 expressed by recombinant adenovirus for esophageal neoplasm target therapy.

Esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EA) are the two most common types of esophageal cancer, which is the sixth highest cause of cancer­associated mortality and the eighth most common cancer worldwide. Gene associated with retinoid­interferon (IFN)­induced mortality­19 (Grim­19) is reported to be a cell death activator that may be used to define mechanisms involved in IFN­ß­ and retinoic acid­induced cell death and apoptosis in a number of tumor cell lines. The present study constructed a recombinant adenovirus expressing Grim­19 (rAd­Grim­19) and investigated its therapeutic outcomes in ESCC cells and tumor­bearing mice. Grim­19 expression was detected in EC­109 (ESCC) cells by reverse transcription­quantitative polymerase chain reaction and western blot analysis. Tumor cell death and apoptosis induced by rAd­Grim­19 in EC­109 cells were analyzed by flow cytometry. The inhibitory effects of rAd­Grim­19 on EC­109 growth were determined by MTT assays. Furthermore, the therapeutic effects of rAd­Grim­19 were investigated in EC­109­bearing mice. The results demonstrated that Grim­19 mRNA and protein expression was downregulated in EC­109 esophageal carcinoma cells compared with Het­1A normal esophageal epithelial cells. In addition, EC­109 cells exhibited a significant reduction in tumor cell growth in the rAd­Grim­19 group compared with the control groups. Furthermore, rAd­Grim­19 increased EC­109 cell apoptosis compared with the control group. These results indicated that rAd-Grim-19 may regulate tumor cell growth and apoptosis. Additionally, the results demonstrated that rAd­Grim­19 led to beneficial outcomes and prolonged the survival of esophageal tumor­bearing mice. In conclusion, the present study demonstrated that rAd­Grim­19 may have potential as an antitumor agent for esophageal neoplasms and may therefore be beneficial for patients with esophageal neoplasms.

Preoperative 3-dimensional computed tomography lung simulation before video-assisted thoracoscopic anatomic segmentectomy for ground glass opacity in lung.

BACKGROUND: Three-dimensional (3D) simulation of pulmonary vessels and the space between the lesion and adjacent tissues may improve the safety and accuracy of video-assisted thoracoscopic surgery (VATS) for lung. The aim of this study was to evaluate the effect of 3D simulation on the outcome of VATS segmentectomy for ground glass opacity (GGO) in lung. METHODS: We retrospectively analyzed 68 cases of small (≤2 cm) GGO, which were diagnosed as cT1aN0M0 lung cancer, from May 1, 2016 to February 28, 2017 in our institute. All the patients underwent VATS segmentectomy. The patients were divided into "3D" group, 3D preoperative reconstruction simulation in 36 patients and "non-3D" group, 32 patients with only computed tomography (CT). Operation plans were firstly made by CT in all patients, then by 3D simulation only in 3D group. The clinical outcomes, including operation time, blood loss, resection margin distance, length of postoperative stay and postoperative complications were compared between the two groups. RESULTS: There were 21 male and 47 female analyzed, aging from 34 to 72 years (median 57). In 3D group, pathological result showed 8 cases of adenocarcinoma, 23 cases of microinvasive adenocarcinoma (MIA), 5 cases of adenocarcinoma in situ (AIS). In non-3D group, 18 cases of MIA, 9 cases of adenocarcinoma and 5 cases of AIS were diagnosed pathologically. The blood loss, postoperative hospital stay and the incidence of the postoperative complications were similar in both of the groups. There was no 30-day postoperative mortality in either group. The median operation time for the 3D group (111 minute) was shorter than non-3D group (139 minute) (P=0.03). Seven cases (19%) in 3D group changed the original operation plan according to the simulation result with the consideration of adequate resection margin distance. All cases in 3D group had adequate resection margin distance. Four cases (13%) in non-3D group got inadequate resection margin distance, and more lung tissues than the original plan were then resected in these patients (P=0.04). CONCLUSIONS: 3D preoperative simulation may be more precise in operation plan than CT scan and can significantly shorten the operation time in VATS segmentectomy for GGO in lung.

Hyperbaric oxygen alleviates the activation of NLRP­3­inflammasomes in traumatic brain injury.

Growing evidence has demonstrated that the nucleotide­binding oligomerization domain­like receptor family pyrin domain containing 3 (NLRP­3) inflammasome­mediated inflammatory pathways have been involved in the secondary injury of traumatic brain injury (TBI). In the present study, the authors investigated the effects of hyperbaric oxygen (HBO) therapy on the NLRP­3 inflammasome pathway following TBI. Following the evaluation of motor deficits and brain edema, the therapeutic effects of HBO on interleukin (IL)­1ß and IL­18 expression were assessed, as well as NLRP­3 inflammasome activation following TBI. HBO may improve motor score and reduce brain edema, accompanied with the reduction of IL­1ß and IL­18 during the 7­day observation period. Furthermore, HBO suppressed mRNA and protein expression of NLRP­3­inflammasome components, especially reducing NLRP­3 expression in microglia. Thus, these results suggested that HBO alleviates the inflammatory response in experimental TBI via modulating microglial NLRP­3­inflammasome signaling.

MACC­1 antibody target therapy suppresses growth and migration of non­small cell lung cancer.

Non­small­cell lung cancer (NSCLC) accounts for ~80% of human lung cancers that result in mortalities worldwide. Metastasis­associated in colon cancer­1 (MACC­1) has been demonstrated to be significantly expressed in cases of NSCLC and promotes tumor cell migration and metastasis through transactivation of the metastasis­inducing hepatocyte growth factor/MET proto­gene, receptor tyrosine kinase (HGF/MET) signaling pathway. The present study constructed a chimeric antibody (Chanti­MACC­1) targeting MACC­1 and investigated its potential as a molecular therapeutic target in the treatment of NSCLC therapy. The expression of MACC­1 was detected by reverse transcription­quantitative polymerase chain reaction and western blotting in lung cancer cell lines and tissues. MTT assay was used to detect proliferation of A549 cells treated by Chanti­MACC­1, whereas the functional and regulatory effects of Chanti­MACC­1 in the migration and metastasis of NSCLC cells was investigated by a cell invasion assay. The therapeutic effect and survival time was observed in animal models. The results demonstrated that MACC­1 expression was increased and overexpression of MACC­1 promoted the progression of the cell cycle, significantly promoted NSCLC cell growth and enhanced tumor migration and invasion through the HGF/MET signaling pathway. It was further demonstrated that Chanti­MACC­1 efficiently suppressed MACC­1 expression and significantly inhibited NSCLC cell proliferation, migration and invasion by blocking the HGF/MET signaling pathway. The data revealed that Chanti­MACC­1 was not only beneficial for tumor remission, however additionally contributed to the long­term survival of NSCLC ­bearing mice. The findings of the present study indicated that MACC­1 was significantly upregulated and promoted tumor cell growth and migration in NSCLC cells and tissues via transactivation of the metastasis­inducing HGF/MET signaling pathway. However, Chanti­MACC­1significantly inhibited tumor growth and metastasis, which suggested that MACC­1 may be essential for tumor initiation and progression by negatively regulating tumor suppressors.

Epidermal growth factor receptor and B7-H3 expression in esophageal squamous tissues correlate to patient prognosis.

Biomarkers that can serve as diagnostic and prognostic indicators of esophageal squamous cell carcinoma (ESCC) are urgently needed to help improve patient outcomes. Here, the expression of epidermal growth factor receptor (EGFR) and costimulatory molecule B7-H3, both of which have been implicated in tumor onset and progression in certain tumors, was investigated in relation to the clinical characteristics and survival outcomes of patients with ESCC. ESCC tissue samples were analyzed for 100 patients. Tumor and patient characteristics were recorded. Tissues were investigated for EGFR and B7-H3 staining by immunohistochemistry. Patients were followed for up to 96 months to determine overall survival (OS) and progression-free survival (PFS). High expression for EGFR (68.0%) and B7-H3 (66.0%) was observed in the majority of cases. High expression of either EGFR or B7-H3 was correlated with tumor invasion depth and clinical stage (P<0.05). Further, high expression of either EGFR or B7-H3 was correlated with worse survival outcomes. The estimated OS (38.1 months) and PFS (13.4 months) of patients with high expression of EGFR were lower than those of patients with low expression (69.3 and 68.1 months, P<0.05). The estimated OS (31.1 months) and PFS (13.1 months) of patients with high expression of B7-H3 were also lower than those of patients with low expression (69.3 and 66.6 months, P<0.05). Indeed, Cox multiple regression showed that OS and PFS were correlated with EGFR (relative risk =1.853, 1.875, respectively) and B7-H3 (relative risk =1.886, 2.061, respectively) (all P<0.05) expression level. Thus, EGFR and B7-H3 are highly expressed in tumor tissues of patients with ESCC. Their expression levels are correlated with tumor severity and survival, and therefore these may be viable biomarkers to aid in prognosis determination.

Meta-analysis of 18FDG PET-CT for nodal staging in patients with esophageal cancer.

BACKGROUND: We aimed to evaluate the value of (18)FDG PET-CT for detection of regional nodal metastasis in patients with esophageal cancer before surgery. METHODS: A computer search about PET-CT original articles was conducted from January 2000 to December 2012. The reference standard was histopathologic analysis. Two reviewers independently searched articles and extracted data. Sensitivity, specificity, diagnostic odds ratio, and likelihood ratio were pooled for PET-CT using bivariate models. Summary receiver operating characteristic (SROC) curves were also used to summarize overall test performance. RESULTS: Across all seven studies on a per-station analysis (2232 stations), the pooled sensitivity and specificity with 95% confidence interval for PET-CT were 0.62 (0.40-0.79) and 0.96 (0.93-0.98). Across all 6 studies on a per-patient analysis (245 patients), corresponding values for PET-CT were 0.55 (0.34-0.74) and 0.76 (0.66-0.83). CONCLUSIONS: (18)FDG PET-CT had lower sensitivity and accuracy for detection of regional nodal metastasis in patients with esophageal cancer before surgery.

Sleeve lobectomy versus pneumonectomy for non-small cell lung cancer: a meta-analysis.

AIM: It is controversial that whether sleeve lobectomy (SL) should be promoted more worthy than pneumonectomy (PN) in suitable patients. METHODS: We searched all studies that had been published in English from PUBMED and Embase which compared the short-term and long-term outcomes of SL and pneumonectomy (PN) in patients with non-small cell lung cancer (NSCLC). RESULTS: Nineteen studies met our criteria with a combined total of 3878 subjects, of which 1316 (33.9%) underwent SL and 2562 (66.1%) underwent PN. The odds ratio was 0.50 (95% CI: 0.34-0.72) for postoperative mortality, 1.17 (95% CI: 0.82-1.67) for postoperative complications, 0.78 (95% CI: 0.47-1.29) for locoregional recurrences. The risk difference for 1-, 3-, 5- year was 0.11 (95% CI: 0.07-0.14), 0.15 (95% CI: 0.06-0.24), 0.15 (95% CI: 0.09-0.20),respectively. The pooled hazard ratio was 0.63 (95% CI: 0.56-0.71) in favor of SL group. CONCLUSION: SL is more worthy to be done than PN in suitable patients with less mortality and better long-term survival.