Intrahepatic cholangiocarcinoma with FGFR alterations: A series of Chinese cases with an emphasis on their clinicopathologic and genetic features.

Intrahepatic Cholangiocarcinoma (iCCA) with FGFR alterations is relatively rare, and its identification is important in the era of targeted therapy. We collected a large series of FGFR-altered cases in the Chinese population and characterized their clinicopathological and genetic features. Among the 18 FGFR-altered cases out of 260 iCCAs, 10 were males and 8 were females, ranging in age from 35 to 74 years (mean, 57.3 years; median, 58 years). Pathologically, they include 9 cases of large duct (LD, 50 %) and small duct (SD, 50 %) types each. All of them (100 %, 18/18) showed microsatellite stable (MSS) and low tumor mutation burden (TMB). Genetically, FGFR alterations involved FGFR1 (20 %), FGFR2 (70 %), and FGFR3 (10 %), with FGFR2 rearrangement accounting for the most (11/18). The most frequently altered genes/biological processes were development/proliferation-related pathways (44 %), chromatin organization (20 %), and tumor suppressors (32 %). Our study further revealed the clinicopathological and genetic features of FGFR-altered iCCA and demonstrated that its occurrence may show regional or ethnic variability and is less common in the Chinese population. A significant number of LD-type iCCA cases also have FGFR alterations rather than the SD type.

Role of micronucleus-activated cGAS-STING signaling in antitumor immunity. / å¾®æ ¸æ¿€æ´»cGAS-STINGä¿¡å·é€šè·¯çš„æœºåˆ¶åŠå ¶è‚¿ç˜¤å ç–«åŠŸèƒ½.

Cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS)-stimulator of interferon genes (STING) signaling is a significant component of the innate immune system and functions as a vital sentinel mechanism to monitor cellular and tissue aberrations in microbial invasion and organ injury. cGAS, a cytosolic DNA sensor, is specialized in recognizing abnormally localized cytoplasmic double-stranded DNA (dsDNA) and catalyzes the formation of a second messenger cyclic-GMP-AMP (cGAMP), which initiates a cascade of type Ⅰ interferon and inflammatory responses mediated by STING. Micronucleus, a byproduct of chromosomal missegregation during anaphase, is also a significant contributor to cytoplasmic dsDNA. These unstable subcellular structures are susceptible to irreversible nuclear envelope rupture, exposing genomic dsDNA to the cytoplasm, which potently recruits cGAS and activates STING-mediated innate immune signaling and its downstream activities, including type Ⅰ interferon and classical nuclear factor-κB (NF-κB) signaling pathways lead to senescence, apoptosis, autophagy activating anti-cancer immunity or directly killing tumor cells. However, sustained STING activation-induced endoplasmic reticulum stress, activated chronic type Ⅰ interferon and nonclassical NF-κB signaling pathways remodel immunosuppressive tumor microenvironment, leading to immune evasion and facilitating tumor metastasis. Therefore, activated cGAS-STING signaling plays a dual role of suppressing or facilitating tumor growth in tumorigenesis and therapy. This review elaborates on research advances in mechanisms of micronucleus inducing activation of cGAS-STING signaling and its implications in tumorigenesis and therapeutic strategies of malignant tumors.

Deletion of cis-regulatory Element in FOXL2 Promoter in a Chinese Family of Type II Blepharophimosis-ptosis-epicanthus Inversus Syndrome with Polydactyly.

Blepharophimosis-ptosis-epicanthus inversus syndrome (BPES) is a relatively uncommon autosomal-dominant genetic disorder, primarily attributed to mutations in the forkhead box L2 (FOXL2) gene. Albeit the involvement of protein-coding regions of FOXL2 has been observed in the majority of BPES cases, whether deficiencies in regulatory elements lead to the pathogenesis remains poorly understood. Herein, an autosomal-dominant BPES type II family was included. Peripheral venous blood has been collected, and genomic DNA has been extracted from leukocytes. A whole exome sequencing analysis has been performed and analyzed (Deposited in NODE database: OER422653). The promoter region of FOXL2 was amplified using polymerase chain reaction (PCR). The luciferase reporter assay was performed to identify the activity of this region. In this study, we present a Chinese family diagnosed with type II BPES, characterized by the presence of small palpebral fissures, ptosis, telecanthus, and epicanthus inversus. Notably, all male individuals within the family display polydactyly. A 225-bp deletion in the 556-bp 5'-upstream to transcription start site of FOXL2 , decorated by multiple histone modifications, was identified in affected members of the family. This deletion significantly decreased FOXL2 promoter activity, as measured by the luciferase assay. Conclusively, a novel 255-bp-deletion of the FOXL2 promoter was identified in Chinese families with BPES. Our results expand the spectrum of known FOXL2 mutations and provide additional insight into the genotype-phenotype relationships of the BPES pathogenesis. In addition, this study indicates the important role of genetic screening of cis-regulatory elements in testing heritable diseases.

Identication and validation of cell senescence biomarkers in idiopathic pulmonary hypertension via integrated transcriptome analyses and machine learning.

BACKGROUND: Idiopathic pulmonary hypertension (IPAH) is a rare and severe disease that affects the pulmonary vasculature. As the diagnosis of IPAH requires invasive right heart catheterization surgery, early detection of this condition is notoriously challenging. Therefore, it is of utmost importance to investigate biomarkers present in peripheral blood that could aid physicians in the early identification and management of IPAH. METHOD: We speculate that cellular senescence may be involved in the occurrence and development of IPAH through various pathways. In this study, we utilized integrated transcriptome analyses and machine learning-based approach to develop a diagnostic model for IPAH cell senescence. To select genetic features, we employed two machine learning algorithms: the Least Absolute Shrinkage and Selection Operator (LASSO) and Random Forest (RF). Additionally, we validated our findings through both external data sets and qRT-PCR experiments. RESULTS: The resulting diagnostic nomogram was able to identify five important biomarkers that can aid in the diagnosis of IPAH, including TNFRSF1B, CCL16, GCLM, IL15, and SOD1. These genes are primarily associated with the immune system, as well as with cell senescence and apoptosis. CONCLUSION: Our study demonstrates the utility of machine learning algorithms in making accurate diagnoses of IPAH, providing clinicians with a more directed approach to the diagnosis and treatment of this disease.

Association Between Intermediate End Points, Progression-free Survival, and Overall Survival in First-line Advanced or Recurrent Endometrial Cancer.

PURPOSE: Advanced/recurrent endometrial cancer is associated with poor long-term outcomes. Clinical studies of novel regimens are ongoing, but given that data on overall survival (OS) take a long time to mature, surrogate end points are often used to support clinical-research interpretation. The aim of this study was to explore the correlation between progression-free survival (PFS)/time to progression (TTP) and OS across multiple time points in the first-line treatment of advanced/recurrent endometrial cancer. METHODS: This study comprised meta-analyses of Phase 2/3 randomized, controlled trials of first-line treatments in patients with advanced primary or first-recurrent endometrial cancer identified via systematic literature review. The strength of the surrogacy relationship was assessed by correlation analyses (estimated with Spearman and Pearson correlation coefficients) and weighted linear regression. FINDINGS: Data from 15 studies were included. PFS and TTP (TTP was reported in one study only) were highly correlated with future OS at multiple time points (Spearman values, 0.83-0.90; Pearson values, 0.86-0.93), suggesting that a change in PFS/TTP would likely be correlated with a change in OS in the same direction. On weighted linear regression, a 10% increase in PFS/TTP probability was significantly associated with a 9.3% to 13.3% increase in the probability of future OS. The strong positive association between PFS/TTP and OS was supported by findings from sensitivity analyses based on identified sources of interstudy heterogeneity. IMPLICATIONS: PFS/TTP is a good potential candidate for predicting long-term OS outcomes in trials of first-line treatment in patients with advanced/recurrent endometrial cancer. The findings from this report may help to inform health-authority and clinical decision makers that PFS/TTP improvements are likely to translate into subsequent OS improvements once data mature.

Outcomes for Dostarlimab and Real-World Treatments in Post-platinum Patients With Advanced/Recurrent Endometrial Cancer: The GARNET Trial Versus a US Electronic Health Record-Based External Control Arm.

Background: Patients with advanced or recurrent endometrial cancer (EC) have limited treatment options following platinum-based chemotherapy and poor prognosis. The single-arm, Phase I GARNET trial (NCT02715284) previously reported dostarlimab efficacy in mismatch repair-deficient/microsatellite instability-high advanced or recurrent EC. Objectives: The objective of this study was to compare overall survival (OS) and describe time to treatment discontinuation (TTD) for dostarlimab (GARNET Cohort A1 safety population) with an equivalent real-world external control arm receiving non-anti-programmed death (PD)-1/PD-ligand (L)1/2 treatments (constructed using data from a nationwide electronic health record-derived de-identified database and applied GARNET eligibility criteria). Methods: Propensity scores constructed from prognostic factors, identified by literature review and clinical experts, were used for inverse probability of treatment weighting (IPTW). Kaplan-Meier curves were constructed and OS/TTD was estimated (Cox regression model was used to estimate the OS-adjusted hazard ratio). Results: Dostarlimab was associated with a 52% lower risk of death vs real-world treatments (hazard ratio, 0.48; 95% confidence interval [CI], 0.35-0.66). IPTW-adjusted median OS for dostarlimab (N=143) was not estimable (95% CI, 19.4-not estimable) versus 13.1 months (95% CI, 8.3-15.9) for real-world treatments (N = 185). Median TTD was 11.7 months (95% CI, 6.0-38.7) for dostarlimab and 5.3 months (95% CI, 4.1-6.0) for the real-world cohort. Discussion: Consistent with previous analyses, patients treated with dostarlimab had significantly longer OS than patients in the US real-world cohort after adjusting for the lack of randomization using stabilized IPTW. Additionally, patients had a long TTD when treated with dostarlimab, suggesting a favorable tolerability profile. Conclusion: Patients with advanced or recurrent EC receiving dostarlimab in GARNET had significantly lower risk of death than those receiving real-world non-anti-PD-(L)1/2 treatments.

When self-assembly meets interfacial polymerization.

Interfacial polymerization (IP) and self-assembly are two thermodynamically different processes involving an interface in their systems. When the two systems are incorporated, the interface will exhibit extraordinary characteristics and generate structural and morphological transformation. In this work, an ultrapermeable polyamide (PA) reverse osmosis (RO) membrane with crumpled surface morphology and enlarged free volume was fabricated via IP reaction with the introduction of self-assembled surfactant micellar system. The mechanisms of the formation of crumpled nanostructures were elucidated via multiscale simulations. The electrostatic interactions among m-phenylenediamine (MPD) molecules, surfactant monolayer and micelles, lead to disruption of the monolayer at the interface, which in turn shapes the initial pattern formation of the PA layer. The interfacial instability brought about by these molecular interactions promotes the formation of crumpled PA layer with larger effective surface area, facilitating the enhanced water transport. This work provides valuable insights into the mechanisms of the IP process and is fundamental for exploring high-performance desalination membranes.

Transcriptomic profiling and differential analysis reveal the renal toxicity mechanisms of mice under cantharidin exposure.

Cantharidin (CTD), extracted from the traditional Chinese medicine mylabris, has shown significant curative effects against a variety of tumors, but its clinical application is limited by its high toxicity. Studies have revealed that CTD can cause toxicity in the kidneys; however, the underlying molecular mechanisms remain unclear. In this study, we investigated the toxic effects in mouse kidneys following CTD treatment by pathological and ultrastructure observations, biochemical index detection, and transcriptomics, and explored the underlying molecular mechanisms by RNA sequencing (RNA-seq). The results showed that after CTD exposure, the kidneys had different degrees of pathological damage, altered uric acid and creatinine levels in serum, and the antioxidant indexes in tissues were significantly increased. These changes were more pronounced at medium and high doses of CTD. RNA-seq analysis revealed 674 differentially expressed genes compared with the control group, of which 131 were upregulated and 543 were downregulated. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses showed that many differentially expressed genes were closely related to the stress response, the CIDE protein family, and the transporter superfamily, as well as the MAPK, AMPK, and HIF-1 pathways. The reliability of the RNA-seq results was verified by qRT-PCR of the six target genes. These findings offer insight into the molecular mechanisms of renal toxicity caused by CTD and provide an important theoretical basis for the clinical treatment of CTD-induced nephrotoxicity.

Role of Bone Morphogenetic Protein 4 in the Inflammation of the Myocardium and Vascular Tissue of Obese Mice.

ABSTRACT: Bone morphogenetic protein 4 (BMP4) is a proinflammatory factor. The expression of BMP4 is reduced in the adipose and enhanced in the myocardium and vascular during obesity. It is possibly involved in the process of inflammatory response of the myocardium and vascular. Obesity, often regarded as a risk factor for cardiovascular diseases, is a kind of inflammatory response. This study aimed to investigate the relationship of BMP4 with obesity and cardiovascular disease. Ob/ob mice were used as the experimental group, and C57BL/6 mice were used as the control group. The two groups were further divided into 2 subgroups based on the mice carrying adenovirus-encoding shRNA for BMP4 or Lac Z genes. The messenger RNA and protein levels of BMP4, interleukin-1ß, and interleukin-9 were significantly higher in the myocardial tissue and aorta of ob/ob+ Lac Z shRNA than those in the other 3 groups, whereas the levels in the ob/ob+ BMP4 shRNA group were significantly decreased and comparable with those in the control groups. BMP4 is significantly upregulated in the myocardial tissue and aorta of obese mice, and this suggests that BMP4 is an risk factor involved in the local inflammatory response.

Neoadjuvant Programmed Cell Death Protein 1 Blockade Combined With Stereotactic Body Radiation Therapy for Stage III(N2) Non-Small Cell Lung Cancer: A Case Series.

The addition of radiotherapy in neoadjuvant chemotherapy did not improve event-free or overall survival in resectable non-small cell lung carcinoma (NSCLC). Neoadjuvant immunotherapy produced major pathologic response(MPR) rate of up to 45%. The potential synergy between radiotherapy and immunotherapy has been described in several studies. We reported outcomes of three cases of stage III/N2 NSCLC treated with neoadjuvant immunotherapy and stereotactic body radiation therapy (SBRT) in a single center. This explanatory trial included treatment-naive patients with stage III resectable NSCLC who received two doses of the programmed cell death protein 1 (PD-1) inhibitor toripalimab after 1 week of receiving SBRT for lung lesions. Thereafter, surgery was planned 4-6 weeks after the second dose. The primary endpoints were safety and feasibility, while the secondary endpoint was the pathologic response rate. Toripalimab combined with SBRT as a neoadjuvant treatment had well-tolerable side effects and did not lead to a delay in surgery. Among the included patients, one achieved pathologic complete response (PCR), one achieved MPR, and one with 20% residual tumor did not achieve MPR. There were few side effects of toripalimab combined with SBRT as a neoadjuvant treatment, and the treatment did not cause a delay in surgery. This study preliminarily explored the outcomes of a new neoadjuvant treatment.

Effects of high-dose all-trans retinoic acid on longitudinal bone growth of young rats.

OBJECTIVE: The signaling axis consisting of GH-IGF1-IGFBP3 is the primary signal taht acts prepubertally to influence height development. Growth plate thinning and even premature closure have been reported in children with tumors treated with retinoid chemotherapy, resulting in long bone dysplasia. Growth failure may occur despite received GH treatment, but the reason is unknown. This study investigate the effect of high-dose all-trans retinoic acid (ATRA) on the development of long bones in growing SD rats. METHODS: A total of 20 three-week-old male SD rats were randomly divided into a control group and an experimental group (n = 10). Rats were treated by gavage with or without high-dose ATRA for 10 days. The body weights of the rats were recorded daily. At the end of the experiment, we measured the length of nose-tail and tibia, stained the tibia and liver for pathological tissue and RT-PCR reaction, and measured the levels of serum GH, IGF1 and IGFBP3, and so on. RESULTS: Compared with controls, experimental rats exhibited reduced body weight and shortened nasal-tail and radial tibial length. Cyp26b1 enzyme activity in the liver was elevated, and histopathological staining revealed that the cartilaginous epiphyseal plate was narrowed, the medullary cavity of trabecular bone was sparse, the number of trabecular bones was decreased, trabecular separation was increased, bone marrow mineralization was enhanced, osteoclastic activity was increased, and circulating GH-IGF1-IGFBP3 levels were decreased. However, RT-PCR reaction results of localized proximal tibiae showed upregulation of IGF1 and downregulation of IGFBP3. CONCLUSIONS: High-dose ATRA intake over a short period of time can reduce GH-IGF1-IGFBP3 levels, affect cartilage and bone homeostasis, and inhibit bone growth in developing animals.

Downregulated microRNA-149-3p triggers malignant development and predicts worse prognosis in oral squamous cell carcinoma.

OBJECTIVE: Accumulating evidence reveals that aberrant expression of microRNAs contributes to the tumorigenesis and development of diverse human cancers. In the current study, we aimed to evaluate the functional role and prognostic value of miR-149-3p in oral squamous cell carcinoma (OSCC). METHODS: Real-time polymerase chain reaction (PCR) analysis was performed to detect the expression of miR-149-3p in 70 OSCC patients (64.10 ± 11.97 years; 31 males and 39 females). The prognostic ability of miR-149-3p in OSCC patients was assessed by Kaplan-Meier survival analysis. Transwell assays and cell adhesion assays were used to investigate the impact of miR-149-3p on cell migration and invasion. The regulation of MMP2 expression by miR-149-3p was determined by real-time PCR, western blotting and dual luciferase reporter assay. RESULTS: Our results revealed a lower level of miR-149-3p in OSCC tissues than in adjacent normal tissues. Downregulation of miR-149-3p was correlated with malignant development and poor outcomes in patients with OSCC. MiR-149-3p repressed the migratory and invasive abilities of OSCC cells. We confirmed that miR-149-3p targeted the 3'-untranslated region of MMP2 mRNA to suppress MMP2 expression. Moreover, the miR-149-3p-mediated decrease in metastasis was reversed by overexpression of MMP2 in OSCC cells. CONCLUSION: Our findings provide an important molecular mechanism by which miR-149-3p inhibits OSCC cell migration and invasion via negative regulation of MMP2 and implicate miR-149-3p as a prospective biomarker and therapeutic target for OSCC.

A Novel Curcumin-Based Drug Powder Inhalation Medicine for Chronic Obstructive Pulmonary Disease.

The dry powder inhaler is a new form of drug delivery that is widely used as an alternative to traditional drug delivery methods, addressing the shortcomings of traditional drug delivery methods and obtaining better therapeutic results. This mode of delivery is also one of the most rational ways to treat pulmonary diseases such as chronic obstructive pulmonary disease (COPD). Curcumin, a natural polyphenol, has been shown to be effective in the treatment of COPD. In this study, different concentrations of curcumin ethanol solution were spray dried with mannitol as a carrier to obtain dry powder particles with different particle size distribution for the preparation of curcumin dry powder inhaler. The solubility and physicochemical properties were further characterized by differential scanning calorimetry (DSC), X-ray diffraction (XRD), and scanning electron microscopy. The characterization results showed that the product obtained in the experiment had reasonable particle size distribution and excellent solubility properties, which were positive for the treatment of COPD or other pulmonary diseases.

MiR-4429 Alleviates Malignant Behaviors of Lung Adenocarcioma Through Wnt/ß-Catenin Pathway.

Lung adenocarcinoma (ADC) is a common subtype of non-small cell lung cancer. MicroRNAs have been reported to be effective biomarkers for diagnosis and an important target for therapy. MiR-4429 is a newly identified miRNA, which can take part in tumor progression as a tumor inhibitor. Moreover, it is an exosomal miRNA that can be taken by lung ADC cell line A549. Nevertheless, its role in lung ADC has been poorly studied. This research discovered that miR-4429 was low expressed in lung ADC cells. MiR-4429 mimics could alleviate the capacities of cell proliferation and metastasis. The mimics are able to reverse epithelial-mesenchymal transition at the same time. Furthermore, it was verified that miR-4429 could bind to ß-catenin and negatively regulate ß-catenin expression. Interestingly, SKL2001 can reverse the role of miR-4429 on tumor. Consequently, miR-4429 can inactivate Wnt/ß-catenin signaling pathway by targeting ß-catenin and prevent oncogene expression in lung ADC cells.

Contrast-enhanced computed tomography prior to percutaneous transthoracic needle biopsy reduces the incidence of hemorrhage.

BACKGROUND: Hemorrhage is the second most common complication of percutaneous transthoracic needle biopsy (PTNB), and at present, there is no effective prevention strategy. Contrast-enhanced computed tomography (CECT) has the advantage of clearly visualizing blood supply within the lesion and aiding in the imaging of blood vessels, which can reduce hemorrhage complicating PTNB. As no large-sample studies were evaluating whether CECT could reduce hemorrhage, we conducted the present retrospective study. METHODS: From November 2011 to February 2016, 1,282 biopsies at Jinling Hospital were retrospectively reviewed; 555 underwent CECT, and 727 underwent non-contrast computed tomography (CT). Factors associated with hemorrhage were defined, and hemorrhage rates were compared between the 2 groups. RESULTS: We found that pre-biopsy CECT was associated with a reduced incidence of biopsy-related hemorrhage compared to non-contrast CT (16.4% vs. 23.1%, P=0.003). Propensity score matching (PSM) analysis also showed that the incidence of hemorrhage in the CECT group was lower than that of the non-contrast CT group at a ratio of 1:1 (P=0.039), 1:2 (P=0.028), or 1:3 (P=0.013). In the multivariate analysis, CECT before PTNB was found to be significantly associated with a reduced risk of hemorrhage [odds ratio (OR): 0.671, 95% confidence interval (CI): 0.499-0.902, P=0.008]. Puncture position, lesion size, depth of needle tract, and the number of punctures were also found to be associated with hemorrhage (all P<0.05). CONCLUSIONS: Compared with non-contrast CT, CECT significantly reduced the risk of post-biopsy pulmonary hemorrhage, which suggests that CECT should be performed before PTNB.

In Vitro Metabolism of E2, G2: Novel Bile Acid-Coupling Camptothecin Analogues, in Rat Liver Microsomes.

BACKGROUND: E2 (Camptothecin - 20 (S) - O- glycine - deoxycholic acid), and G2 (Camptothecin - 20 (S) - O - acetate - deoxycholic acid) are two novel bile acid-derived camptothecin analogues, modified deoxycholic acid at 20-position of CPT(camptothecin) with greater anticancer activity and lower systematic toxicity in vivo. OBJECTIVE: We aimed to investigate the metabolism of E2 and G2 by Rat Liver Microsomes (RLM). METHODS: Phase I and Phase II metabolism of E2 and G2 in rat liver microsomes were performed, respectively, and the mixed incubation of phase I and phase II metabolism of E2 and G2 was also processed. Metabolites were identified by liquid chromatographic/mass spectrometry. RESULTS: The results showed that phase I metabolism was the major biotransformation route for both E2 and G2. The isoenzyme involved in their metabolism had some difference. The intrinsic clearance of G2 was 174.7 mL/min. mg protein, more than three times that of E2 (51.3 mL/min . mg protein), indicating a greater metabolism stability of E2. 10 metabolites of E2 and 14 metabolites of G2 were detected, including phase I metabolites (mainly via hydroxylations and hydrolysis) and their further glucuronidation products. CONCLUSION: These findings suggested that E2 and G2 have similar biotransformation pathways except for some differences in the hydrolysis ability of the ester bond and amino bond from the parent compounds, which may result in the diversity of their metabolism stability and responsible CYPs(Cytochrome P450 proteins).

Myosin18B predicts favorable prognosis of cutaneous squamous-cell carcinoma.

BACKGROUND: Myosin18 family, including Myosin18A (MYO18A) and Myosin18B (MYO18B), are newly-identified Myosins in Myosin superfamily. The expression and function of Myosin18 family in cancer progression is still controversial, and in cutaneous squamous-cell carcinoma (cSCC) is totally unknown. OBJECTIVE: To investigate the expression and prognostic significance of Myosin18 family in cSCC. METHODS: In this study, the expressions of MYO18 family, including MYO18A and MYO18B were detected in six pairs of cSCCs and corresponding normal tissues with qRT-PCR. MYO18A and MYO18B expressions and intracellular locations in 80 cSCCs were detected with immunohistochemistry. The clinical significance was evaluated by analyzing the correlation between MYO18 family and clinicopathological factors. The prognostic significance of MYO18 family was estimated by univariate analysis with log-rank test, and by multivariate analysis by Cox-regression model. RESULTS: The percentages of high MYO18A and MYO18B in cSCC were 43.75% and 36.25%, respectively. High expression of MYO18A (P = 0.035) and MYO18B (P = 0.032) were both associated with less tumor size. MYO18A had no significant influence on sSCC prognosis (P = 0.686), but low expression of MYO18B was proved to be significantly associated with poor outcome of cSCC (P = 0.014). MYO18B was confirmed as an independent prognostic biomarker of cSCC (P = 0.002), indicating the favorable outcome. CONCLUSION: The expression of MYO18B was an independent prognostic biomarker of cSCC, predicting the favorable prognosis independently. Investigating the expression of MYO18B can help stratify the subset of high-risk cSCC patients for more potent treatment and post-operational surveillance.

Mutual dependency between lncRNA LETN and protein NPM1 in controlling the nucleolar structure and functions sustaining cell proliferation.

Fundamental processes such as ribosomal RNA synthesis and chromatin remodeling take place in the nucleolus, which is hyperactive in fast-proliferating cells. The sophisticated regulatory mechanism underlying the dynamic nucleolar structure and functions is yet to be fully explored. The present study uncovers the mutual functional dependency between a previously uncharacterized human long non-coding RNA, which we renamed LETN, and a key nucleolar protein, NPM1. Specifically, being upregulated in multiple types of cancer, LETN resides in the nucleolus via direct binding with NPM1. LETN plays a critical role in facilitating the formation of NPM1 pentamers, which are essential building blocks of the nucleolar granular component and control the nucleolar functions. Repression of LETN or NPM1 led to similar and profound changes of the nucleolar morphology and arrest of the nucleolar functions, which led to proliferation inhibition of human cancer cells and neural progenitor cells. Interestingly, this inter-dependency between LETN and NPM1 is associated with the evolutionarily new variations of NPM1 and the coincidental emergence of LETN in higher primates. We propose that this human-specific protein-lncRNA axis renders an additional yet critical layer of regulation with high physiological relevance in both cancerous and normal developmental processes that require hyperactive nucleoli.

MicroRNA­149­3p inhibits cell proliferation by targeting AKT2 in oral squamous cell carcinoma.

MicroRNAs (miRs) exhibit oncogenic or tumor suppressive functions that contribute to the initiation and development of various types of human cancer. miR­149­3p has been reported to serve multiple roles in the regulation of proliferation, apoptosis and metastasis. However, the effects and detailed mechanism of miR­149­3p in oral squamous cell carcinoma (OSCC) remain unclear. In the present study, miR­149­3p mimic, mimic control, miR­149­3p inhibitor and inhibitor control were transiently transfected into Cal27 and SCC­9 cells. The viability, proliferation and apoptosis of OSCC cells were determined using Cell Counting Kit­8, colony formation and Annexin V assays, respectively. The mRNA expression levels of miR­149­3p and AKT2 were determined by reverse transcription­quantitative PCR. The protein expression levels of AKT2, cleaved caspase­3 and cleaved PARP were examined by western blot analysis. The binding of miR­149­3p to the AKT2 3'­untranslated region was evaluated by a dual luciferase reporter assay. In the present study, overexpression of miR­149­3p reduced the viability and proliferation of OSCC cells. By contrast, increased cell viability and proliferation was observed in miR­149­3p­deficient OSCC cells. Dual luciferase reporter assay indicated that miR­149­3p significantly decreased the luciferase activity of the wild­type AKT2 3'­untranslated region. Moreover, overexpression of miR­149­3p downregulated the mRNA and protein expression levels of AKT2, suggesting that miR­149­3p was a negative modulator of AKT2. Restoration of AKT2 efficiently reversed the miR­149­3p­mediated reduction in the proliferative capacity of OSCC cells. In addition, miR­149­3p enhanced the sensitivity of OSCC cells to the chemotherapeutic drug 5­fluorouracil. Taken together, the current findings revealed an inhibitory effect of miR­149­3p on the proliferation of OSCC cells through the post­transcriptional suppression of AKT2, and indicated a potential chemosensitizing function of miR­149­3p for the treatment of patients with OSCC.

MiR-320a inhibits malignant phenotype of melanoma cells via targeting PBX3.

PURPOSE: To study the expression of micro ribonucleic acid-320a (miR-320a) in melanoma cells and its influence on the biological functions of these cells. METHODS: MiR-320a expression data and clinical data in melanoma tissues were downloaded from The Cancer Genome Atlas (TCGA) database. Real time-quantitative polymerase chain reaction (RT-qPCR) was used to detect miR-320a expression in melanoma tissues, malignant melanoma cell lines (A375, SKMEL-28 and A2058) and human epidermal melanoma (HEM) cells. The miR-320a mimic was transfected into A375 cells, and the functions of cells were detected. The luciferase reporter gene assay was employed to verify the miR-320a downstream target protein predicted by the biological information prediction software. RESULTS: The differential analysis of miRNAs in melanoma tissues from TCGA database showed that miR-320a expression in melanoma tissues was significantly lower than that in adjacent tissues, and low expression of miR-320a exhibited a severe poor prognosis (p<0.01). MiR-320a mimic could significantly enhance the expression level of miR-320a (p<0.01). The absorbance at 490 nm of A375 cells overexpressing miR-320a decreased remarkably and their proliferation ability was weakened (p<0.01). Overexpression of miR-320a in A375 cells inhibited cell migration to wound parts and epithelial-mesenchymal transition (EMT), invading malignant phenotype (p<0.05). Flow cytometry was employed and it was denoted that after transfecting with miR-320a, the apoptosis rate of A375 cells was elevated overtly (p<0.01). The dual luciferase report test indicated that the luciferase activity in wild type pre-B-cell leukemia transcription factor 3 (PBX3) was markedly lower than that of mutant type PBX 3 (p<0.05). CONCLUSIONS: Targeted binding of miR-320a to PBX3 protein can inhibit the malignant phenotype of cells and affects the occurrence and development of melanoma.