LncRNA HOTTIP regulates TLR4 promoter methylation by recruiting H3K4 methyltransferase MLL1 to affect apoptosis and inflammatory response of fibroblast-like synoviocyte in rheumatoid arthritis.

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease, and the role of HOXA transcript at the distal tip (HOTTIP) in its pathogenesis remains underexplored. This study investigates the mechanism by which HOTTIP influences apoptosis and the inflammatory response of fibroblast-like synoviocytes (FLS). An RA mouse model was established, and clinical scores were analyzed. Pathological changes in synovial tissues, bone mineral density (BMD) of the paws, serum tartrate-resistant acid phosphatase (TRAP) activity, and TNF-α and IL-1ß levels were assessed. FLS were transfected, and cell proliferation and apoptosis were examined. The RNA-pull-down assay determined HOTTIP's interaction with mixed-lineage leukemia 1 (MLL1), while RNA immunoprecipitation assay measured HOTTIP expression pulled down by MLL1. The levels of MLL1 and toll-like receptor 4 (TLR4) after MLL1 overexpression based on HOTTIP silencing were determined. Chromatin immunoprecipitation (ChIP) was performed with H3K4me3 as an antibody, followed by the evaluation of TLR4 expression. HOTTIP expression was elevated in RA mouse synovial tissues. Inhibition of HOTTIP led to reduced clinical scores, inflammatory infiltration, synovial hyperplasia, TRAP activity, and TNF-α and IL-1ß levels, along with increased BMD. In vitro Interference with HOTTIP suppressed RA-FLS apoptosis and inflammation. HOTTIP upregulated TLR4 expression by recruiting MLL1 to facilitate TLR4 promoter methylation. MLL1 overexpression reversed HOTTIP silencing-mediated repression of RA-FLS apoptosis. Activation of H3K4 methylation counteracted HOTTIP knockout, ameliorating the inflammatory response. HOTTIP regulates TLR4 expression by recruiting MLL1, leading to TLR4 promoter methylation, thereby suppressing RA-FLS proliferation and inducing cell apoptosis and inflammatory response in RA.

Miniaturization of flexible ureteroscopes: a comparative trend analysis of 59 flexible ureteroscopes.

The purpose of this review is to analyze the trend in miniaturization of flexible ureteroscopes over the past decades, identify the advantages and disadvantages, and determine the correlation of individual parameters with release period. Flexible ureteroscopes mentioned in the literature or those commercially available were searched. To minimize the search bias, the instruments were grouped by release date time periods of < 2000 year, 2000-2009, 2010-2019, and 2020 onwards. The final review included only those instrument models for which data on tip size, overall shaft, working length and channel size had been determined. The correlation among features investigated as well as with release period was also determined. 59 models of flexible ureteroscopes (26 fiber optic and 33 digital scopes) were included. Among the different features investigated among fiber optic endoscopes, only the sizes of the distal tip and overall shaft positively correlated with each other. In contrast to their fiber optic counterparts, a strong positive correlation was observed between tip and channel sizes, whereas negative correlation was found between channel size and overall shaft size and working length of digital scopes. Only distal tip of fiber optic flexible ureteroscopes and overall shaft of digital endoscopes were significantly reduced over their evolution. With the development of technology, there has been an improvement of flexible ureteroscopes and one of the indicators of this trend is a decrease in their size. With a definite trend towards miniaturization over the past decades, a significant correlation was observed in tip size and overall shaft for fiber optic and digital endoscopes, respectively.

Influence of maxillary molar distalization with clear aligners on three-dimensional direction: molar distal movement, intrusion, distal tip and crown buccal torque.

BACKGROUND: The aim of this study was to evaluate the distal movement, vertical movement, distal tipping and crown buccal torque of maxillary molars after the completion of distalization by comparing the predicted movement with the achieved movement using palatal rugae registration. METHODS: The study included 22 clear aligner patients (7 males and 15 females), and 79 molars were measured. Two digital models were generated before treatment and after molar distalization and were superimposed after selecting the palatal rugae area for registration in GOM inspect suite software 2022 (GOM; Braunschweig, Germany). The predicted and achieved movements of molar distalization, intrusion, distal tip and crown buccal torque were measured and compared. RESULT: The achieved distalization (1.25 ± 0.79 mm vs. 2.17 ± 1.03 mm, P < 0.001; 1.41 ± 1.00 mm vs. 2.66 ± 1.15 mm, P < 0.001), intrusion (0.47 ± 0.41 mm vs. 0.18 ± 0.54 mm, P < 0.01; 0.58 ± 0.65 mm vs. 0.10 ± 1.12 mm, P < 0.01), distal tip (5.30 ± 4.56° vs. 1.53 ± 2.55°, P < 0.001; 4.87 ± 4.50° vs. - 1.95 ± 4.32°, P < 0.001) and crown buccal torque (1.95 ± 4.18° vs. - 1.15 ± 4.75°, P < 0.001; 0.43 ± 4.39° vs. - 4.27 ± 6.42°, P < 0.001) were significantly different from the predicted values in the two groups (first molar, second molar). Significant regression relationships were found between the achieved distal movement and deviational intrusion (R2 = 0.203, P < 0.0001), distal tip (R2 = 0.133, P < 0.001) and crown buccal torque (R2 = 0.067, P < 0.05). There was a significant correlation between the deviational movements of intrusion and the distal tip (R = 0.555, P < 0.0001). CONCLUSION: Approximately 2 mm maxillary molar distalization was achieved in this study. Deviational movement of intrusion, distal tip and crown buccal torque beyond the clear aligner virtual design appeared to a certain degree after distalization. Thus, more attention should be given to molar intrusion and distal tip and crown buccal torque as the designed distalization increases.

Cep104 is a component of the centriole distal tip complex that regulates centriole growth and contributes to Drosophila spermiogenesis.

Proper centrosome number and function relies on the accurate assembly of centrioles, barrel-shaped structures that form the core duplicating elements of the organelle. The growth of centrioles is regulated in a cell cycle-dependent manner; while new daughter centrioles elongate during the S/G2/M phase, mature mother centrioles maintain their length throughout the cell cycle. Centriole length is controlled by the synchronized growth of the microtubules that ensheathe the centriole barrel. Although proteins exist that target the growing distal tips of centrioles, such as CP110 and Cep97, these proteins are generally thought to suppress centriolar microtubule growth, suggesting that distal tips may also contain unidentified counteracting factors that facilitate microtubule polymerization. Currently, a mechanistic understanding of how distal tip proteins balance microtubule growth and shrinkage to either promote daughter centriole elongation or maintain centriole length is lacking. Using a proximity-labeling screen in Drosophila cells, we identified Cep104 as a novel component of a group of evolutionarily conserved proteins that we collectively refer to as the distal tip complex (DTC). We found that Cep104 regulates centriole growth and promotes centriole elongation through its microtubule-binding TOG domain. Furthermore, analysis of Cep104 null flies revealed that Cep104 and Cep97 cooperate during spermiogenesis to align spermatids and coordinate individualization. Lastly, we mapped the complete DTC interactome and showed that Cep97 is the central scaffolding unit required to recruit DTC components to the distal tip of centrioles.

Organoid modeling of human fetal lung alveolar development reveals mechanisms of cell fate patterning and neonatal respiratory disease.

Variation in lung alveolar development is strongly linked to disease susceptibility. However, underlying cellular and molecular mechanisms are difficult to study in humans. We have identified an alveolar-fated epithelial progenitor in human fetal lungs, which we grow as self-organizing organoids that model key aspects of cell lineage commitment. Using this system, we have functionally validated cell-cell interactions in the developing human alveolar niche, showing that Wnt signaling from differentiating fibroblasts promotes alveolar-type-2 cell identity, whereas myofibroblasts secrete the Wnt inhibitor, NOTUM, providing spatial patterning. We identify a Wnt-NKX2.1 axis controlling alveolar differentiation. Moreover, we show that differential binding of NKX2.1 coordinates alveolar maturation, allowing us to model the effects of human genetic variation in NKX2.1 on alveolar differentiation. Our organoid system recapitulates key aspects of human fetal lung stem cell biology allowing mechanistic experiments to determine the cellular and molecular regulation of human development and disease.

PI3K signaling specifies proximal-distal fate by driving a developmental gene regulatory network in SOX9+ mouse lung progenitors.

The tips of the developing respiratory buds are home to important progenitor cells marked by the expression of SOX9 and ID2. Early in embryonic development (prior to E13.5), SOX9+progenitors are multipotent, generating both airway and alveolar epithelium, but are selective progenitors of alveolar epithelial cells later in development. Transcription factors, including Sox9, Etv5, Irx, Mycn, and Foxp1/2 interact in complex gene regulatory networks to control proliferation and differentiation of SOX9+progenitors. Molecular mechanisms by which these transcription factors and other signaling pathways control chromatin state to establish and maintain cell-type identity are not well-defined. Herein, we analyze paired gene expression (RNA-Seq) and chromatin accessibility (ATAC-Seq) data from SOX9+ epithelial progenitor cells (EPCs) during embryonic development in Mus musculus. Widespread changes in chromatin accessibility were observed between E11.5 and E16.5, particularly at distal cis-regulatory elements (e.g. enhancers). Gene regulatory network (GRN) inference identified a common SOX9+ progenitor GRN, implicating phosphoinositide 3-kinase (PI3K) signaling in the developmental regulation of SOX9+ progenitor cells. Consistent with this model, conditional ablation of PI3K signaling in the developing lung epithelium in mouse resulted in an expansion of the SOX9+ EPC population and impaired airway epithelial cell differentiation. These data demonstrate that PI3K signaling is required for epithelial patterning during lung organogenesis, and emphasize the combinatorial power of paired RNA and ATAC seq in defining regulatory networks in development.

Studying how lungs develop has helped us understand and treat often-devastating lung diseases. This includes diseases like cystic fibrosis which result from spelling mistakes known as mutations in a person's genetic code. However, not all lung diseases involve mutations. Many other diseases, in both adults and children, are caused by genes failing to switch on or off at some point during lung development. DNA is surrounded by various proteins which package it into a compressed structure known as chromatin. Cells can control which genes are turned on or off by modifying how tightly packed parts of the genetic code are within chromatin. Changes in chromatin accessibility, also known as 'epigenetic' changes, are a normal part of development, and guide cells towards specific jobs or identities as an organ matures. However, how this happens in the developing lung is poorly understood. Here, Khattar, Fernandes et al. set out to determine how chromatin accessibility shapes development of the tissue lining the lungs, focusing on a group of progenitor cells which produce the protein SOX9. These cells are initially found at the tips of the early lung, where they go on to develop into the cells that line the whole of the mature organ. Initial experiments used large-scale genetic techniques to measure gene activity and chromatin accessibility simultaneously in progenitor cells extracted from the lungs of mice. Khattar, Fernandes et al. were then able to predict the signaling pathways that shape the lung lining based on which genes were surrounded by unpacked chromatin, and determine the proteins responsible for these epigenetic changes. This included the signaling pathway Phosphatidylinositol 3 kinase (PI3K) which is involved in a number of cellular processes. Additional experiments in mice confirmed that the PI3K pathway became active very early in lung development and remained so until adulthood. In contrast, mice lacking a gene that codes for a key part of the PI3K pathway had defective lungs which failed to develop a proper lining. The data generated in this study will provide an important resource for future studies investigating how epigenetic changes drive normal lung development. Khattar, Fernandes et al. hope that this knowledge will help researchers to better understand the cause of human lung diseases, and identify already available 'epigenetic drugs' which could be repurposed to treat them.

Long-noncoding RNA HOXA transcript at the distal tip ameliorates the insulin resistance and hepatic gluconeogenesis in mice with gestational diabetes mellitus via the microRNA-423-5p/wingless-type MMTV integration site family member 7A axis.

Long-noncoding RNA HOXA transcript at the distal tip (HOTTIP) has been probed to exert essential effects on diabetes progression, while its function in gestational diabetes mellitus (GDM) remains unclear. This study was committed to unravel the effects of HOTTIP on GDM progression via the microRNA (miR)-423-5p/wingless-type MMTV integration site family member 7A (WNT7A) axis. The GDM mouse model was established. HOTTIP, miR-423-5p and WNT7A levels in GDM mice were examined. The saline with dissolved various constructs altering HOTTIP, miR-423-5p and WNT7A expression was injected into GDM mice to detect the levels of GDM-related biochemical indices, HOMA indices, liver gluconease: expression levels of phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G-6-Pase), glucose transporter 2 (GLUT2) and pathological changes of pancreatic tissues, and the apoptosis rate of pancreatic cells in GDM mice. The relations among HOTTIP, miR-423-5p and WNT7A were validated. HOTTIP and WNT7A levels were decreased while miR-423-5p was elevated in GDM mice. The enriched HOTTIP or silenced miR-423-5p alleviated the levels of GDM-relatedbiochemical indices, enhanced the insulin homeostasis, elevated GLUT2 expression and decreased G-6-pase and PEPCK expression, mitigated the pathological changes of pancreatic tissues, and hindered the apoptosis of pancreatic cells. MiR-143-5p upregulation abrogated the effects of elevated HOTTIP on repressing GDM; whereas WNT7A deletion reversed the therapeutic effects of reduced miR-423-5p. HOTTIP sponged miR-423-5p that targeted WNT7A. HOTTIP ameliorates insulin resistance and hepatic gluconeogenesis in GDM mice via the modulation of the miR-423-5p/WNT7A axis. This study affords novel therapeutic modalities for GDM treatment.

An outcome comparison of superficial versus deep plane dissection of cervicofacial flaps: A systematic review and meta-analysis.

PURPOSE: Cervicofacial flaps represent an excellent option for coverage of cheek defects secondary to oncologic resection, trauma or infection. However, there remains clinical equipoise regarding whether superficial plane or deep plane dissection results in the lowest rates of complications and optimal outcomes. METHODS: A systematic review and meta-analysis of proportions was conducted to assesses outcomes between cheek reconstruction superficial plane or deep plane cervicofacial flaps. Outcome measures included flap necrosis, ectropion, hematoma formation, facial nerve injury, and requirement for further operative or non-operative intervention. RESULTS: Of 881 citations identified for review, 10 met the inclusion criteria. In total, 284 patients received superficial plane flaps while 44 patients received deep plane flaps. Overall, reported rates of complications were low for cervicofacial flaps. The proportion of necrosis, ectropion, and hematoma were 3.05% (95% CI: 0.00-10.71%), 2.03% (95% CI: 0.41-4.42%), and 0.05% (95% CI: 0.00-3.29%), respectively. No cases of permanent facial nerve injury were reported. Sub-group analysis demonstrated comparable rates of complications between superficial and deep plane dissection and no difference was found between groups. Other complications were noted with low incidence. CONCLUSIONS: Currently published literature demonstrates that superficial and deep plane cervicofacial flaps exhibit similar rates of complications, although there is a low level of evidence overall. Overall, the rates of flap necrosis (3.05%), ectropion (2.03%), and hematoma (0.05%) are low. Notably, there were no reported cases of permanent facial nerve injury from either technique.

The apical cell - An enigmatic somatic cell in leech ovaries - Structure and putative functions.

Although somatic cells play an integral role in animal gametogenesis, their organization and function are usually poorly characterized, especially in non-model systems. One such example is a peculiar cell found in leech ovaries - the apical cell (AC). A single AC can be found at the apical tip of each ovary cord, the functional unit of leech ovaries, where it is surrounded by other somatic and germline cells. The AC is easily distinguished due to its enormous size and its numerous long cytoplasmic projections that penetrate the space between neighboring cells. It is also characterized by a prominent accumulation of mitochondria, Golgi complexes and electron-dense vesicles. ACs are also enriched in cytoskeleton, mainly in form of intermediate filaments. Additionally, the AC is connected to neighboring cells via junctions that structurally resemble hemidesmosomes. In spite of numerous descriptive data about the AC, its functions remain poorly understood. Its suggested functions include a role in forming skeleton for the germline cells, and a role in defining a niche for germline stem cells. The latter is more speculative, since germline stem cells have not been identified in leech ovaries. Somatic cells with similar morphological properties to those of the AC have been found in gonads of nematodes - the distal tip cell - and in insects - Verson's cell, hub cells and cap cells. In the present article we summarize information about the AC structure and its putative functions. AC is compared with other well-described somatic cells with potentially similar roles in gametogenesis.

Our emerging understanding of the roles of long non-coding RNAs in normal liver function, disease, and malignancy.

Long non-coding RNAs (lncRNAs) are important biological mediators that regulate numerous cellular processes. New experimental evidence suggests that lncRNAs play essential roles in liver development, normal liver physiology, fibrosis, and malignancy, including hepatocellular carcinoma and cholangiocarcinoma. In this review, we summarise our current understanding of the function of lncRNAs in the liver in both health and disease, as well as discuss approaches that could be used to target these non-coding transcripts for therapeutic purposes.

More than just a pool.

An intricate stem cell niche boundary formed by finger-like extensions generates asymmetry in stem cell divisions.

Extension Mechanism of a Flexible End for Colonoscopy.

INTRODUCTION: The significance of the distal tip extension mechanism (DTEM) arises when the insertion tube of the colonoscope is no longer able to move further inside the colon, and when a longer insertion tube is needed. The main focus of this research is to investigate the development of a novel distal tip extension mechanism (DTEM). METHODS: To characterize the performance of the DTEM, the ability of the DTEM to extend the distal tip of the colonoscope 16 cm is investigated. To determine the maximum number of turns that the extension knob needs to make in order to move the distal tip 16 cm, the DTEM is used to displace the distal tip in different colon configurations using the silicon tube of a colonoscopy training model (CTM). The experimentally collected data was presented and discussed to characterize the performance of the DTEM. RESULTS: The results showed that the DTEM is able to extend the distal tip 16 cm while the colon is in various configurations. Additionally, the impact of implanting the DTEM on the flexibility of the insertion tube was determined. DISCUSSION: The results of this research suggest that the DTEM will be an effective tool to help colonoscopists performing better colonoscopies.

Stem cell niche exit in C. elegans via orientation and segregation of daughter cells by a cryptic cell outside the niche.

Stem cells reside in and rely upon their niche to maintain stemness but must balance self-renewal with the production of daughters that leave the niche to differentiate. We discovered a mechanism of stem cell niche exit in the canonical C. elegans distal tip cell (DTC) germ stem cell niche mediated by previously unobserved, thin, membranous protrusions of the adjacent somatic gonad cell pair (Sh1). A disproportionate number of germ cell divisions were observed at the DTC-Sh1 interface. Stem-like and differentiating cell fates segregated across this boundary. Spindles polarized, pairs of daughter cells oriented between the DTC and Sh1, and Sh1 grew over the Sh1-facing daughter. Impeding Sh1 growth by RNAi to cofilin and Arp2/3 perturbed the DTC-Sh1 interface, reduced germ cell proliferation, and shifted a differentiation marker. Because Sh1 membrane protrusions eluded detection for decades, it is possible that similar structures actively regulate niche exit in other systems.

Stem cells have the rare ability to divide and specialize into the many different types of cells necessary for an organism to survive. For instance, germ stem cells can multiply to produce precursor cells that go on to become eggs or sperm needed for reproduction. When a stem cell divides, the daughter cells can either remain 'naïve', or start to specialize into a given cell type. In many cases, this decision is strongly influenced by the properties of the environment that surrounds the stem cell. However, in the microscopic worm Caenorhabditis elegans, how the daughters of germ stem cells specialize was thought to be a random process, with nearby cells equally likely to specialize or remain naïve. In this animal, germ stem cells reside in tube-shaped structures called gonads, which are enclosed by a large 'distal tip' cell. In addition, cells known as Sh1 surround the gonad. Here, Gordon et al. tracked dividing germ stem cells in the gonads of live worms. This revealed that both the distal tip cell and Sh1 cells have finger-like extensions that form contacts with the germ stem cells. The fate of dividing germ stem cells is shaped by these interactions. If they touch only the distal tip cell, they remain in a naïve state. However, if they contact both the distal tip cell and an Sh1 cell, one daughter of the stem cell becomes an egg precursor ­ with the daughter closest to the distal tip cell staying naïve. In fact, germ stem cells that are prevented from contacting Sh1 cells divide less often. Many other types of stem cells, for example in human skin, are believed to make the decision to remain naïve or undergo specialization randomly. The results from Gordon et al. could provide a roadmap to discover hidden layers of control in other organisms, some of which may be potentially relevant in health and disease.

The Caenorhabditis elegans homolog of human copper chaperone Atox1, CUC-1, aids in distal tip cell migration.

Cell migration is a fundamental biological process involved in for example embryonic development, immune system and wound healing. Cell migration is also a key step in cancer metastasis and the human copper chaperone Atox1 was recently found to facilitate this process in breast cancer cells. To explore the role of the copper chaperone in other cell migration processes, we here investigated the putative involvement of an Atox1 homolog in Caenorhabditis elegans, CUC-1, in distal tip cell migration, which is a key process during the development of the C. elegans gonad. Using knock-out worms, in which the cuc-1 gene was removed by CRISPR-Cas9 technology, we probed life span, brood size, as well as distal tip cell migration in the absence or presence of supplemented copper. Upon scoring of gonads, we found that cuc-1 knock-out, but not wild-type, worms exhibited distal tip cell migration defects in approximately 10-15% of animals and, had a significantly reduced brood size. Importantly, the distal tip cell migration defect was rescued by a wild-type cuc-1 transgene provided to cuc-1 knock-out worms. The results obtained here for C. elegans CUC-1 imply that Atox1 homologs, in addition to their well-known cytoplasmic copper transport, may contribute to developmental cell migration processes.

The HOTTIP (HOXA transcript at the distal tip) lncRNA: Review of oncogenic roles in human.

HOXA transcript at the distal tip (HOTTIP) is a 3764 nucleotide long non-coding RNA (lncRNA) encoded from a genomic region in the 5' tip of the HOXA locus. This lncRNA has a role in transmission of signals from higher order chromosomal configuration into chromatin codes. HOTTIP directly binds with the WDR5 protein and recruits the WDR5/MLL complexes across the HOXA locus which leads to H3K4 methylation and activation of the transcription of HOXA genes. This lncRNA has a prominent role in the pathogenesis of almost all kinds of cancers. Apart from a single study in glioma cells, all in vitro studies have emphasized on oncogenic roles of HOTTIP in different malignancies. In vivo studies also showed the effect of HOTTIP silencing in reduction of tumorigenicity in all cancer types except from glioma. Results of clinical studies mostly demonstrated up-regulation of this lncRNA in cancer samples compared with non-malignant tissues of the same origin and correlation between its expression levels and patients' outcome. Taken together, HOTTIP is regarded as an oncogenic lncRNA in almost all kinds of cancers and a putative biomarker and therapeutic target in human malignancies.

LncRNA HOTTIP facilitates the stemness of breast cancer via regulation of miR-148a-3p/WNT1 pathway.

Emerging evidence suggests that dysregulation of long non-coding RNA (lncRNA) plays a key role in tumorigenesis. The lncRNA, HOXA transcript at the distal tip (HOTTIP), has been reported to be up-regulated in multiple cancers, including breast cancer, and is involved in various biological processes, including the maintenance of stemness. However, the biological function and underlying modulatory mechanism of HOTTIP in breast cancer stem cells (BCSCs) remains unknown. In this study, we found that HOTTIP was markedly up-regulated in BCSCs and had a positive correlation with breast cancer progression. Functional studies revealed that overexpression of HOTTIP markedly promoted cell clonogenicity, increased the expression of the stem cell markers, OCT4 and SOX2, and decreased the expression of the differentiation markers, CK14 and CK18, in breast cancer cells. Knockdown of HOTTIP inhibited the CSC-like properties of BCSCs. Consistently, depletion of HOTTIP suppressed tumour growth in a humanized model of breast cancer. Mechanistic studies demonstrated that HOTTIP directly binds to miR-148a-3p and inhibits the mediation of WNT1, which leads to inactivation of the Wnt/ß-catenin signalling pathway. Our study is the first to report that HOTTIP regulates the CSC-like properties of BCSCs by as a molecular sponge for miR-148a-3p to increase WNT1 expression, offering a new target for breast cancer therapy.

Long non-coding (lnc)RNA PAPAS overexpression inhibits tumor growth in papillary thyroid carcinoma by downregulating lncRNA HOTTIP.

The long non-coding (lnc)RNA pre-ribosomal RNA antisense transcripts (PAPAS) can repress ribosomal RNA synthesis, although its role in human disease is currently unknown. The present study aimed to investigate the function of PAPAS in papillary thyroid carcinoma (PTC). PAPAS was downregulated, while the lncRNA HOXA transcript at the distal tip (HOTTIP) was upregulated in patients with PTC. PAPAS and HOTTIP were inversely associated in PTC. PAPAS overexpression led to the downregulation of HOTTIP in PTC cells, while PAPAS expression was not significantly affected by HOTTIP overexpression, suggesting that PAPAS was upstream of HOTTIP. PAPAS expression level decreased, while HOTTIP expression level increased with the increase of clinical staging. PAPAS overexpression led to the inhibition of cell proliferation, while HOTTIP overexpression increased proliferation of PTC cells, and HOTTIP overexpression decreased the effects of PAPAS overexpression. lncRNA PAPAS overexpression may inhibit tumor growth in papillary thyroid carcinoma by downregulating lncRNA HOTTIP.

Expression and clinical significance of long non-coding RNA HOTTIP in tissues of patients with endometrial carcinoma / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression and clinical significance of long non-coding RNA (lncRNA) HOTTIP in tissues of patients with endometrial carcinoma. Methods: A total of 109 cases of patients with endometrial carcinoma who underwent surgery in Xingxiang Central Hospital from April 2012 to April 2014 were selected. The endometrial carcinoma tissue and its corresponding adjacent tissue (more than 5 cm from the cancer margin) were obtained. The expressions of HOTTIP in endometrial carcinoma and adjacent tissues were detected by qRT-PCR. All patients were followed up from the first postoperative day. The follow-up deadline was April 30, 2019. The end-point event was death and the patient's survival time was recorded. Results: The relative expression level of HOTTIP in endometrial carcinoma tissues was (2.55±0.21), which was higher than that in the adjacent tissue (1.03±0.16) (t=60.631, P<0.01). The differences of the relative expression levels of HOTTIP in endometrial carcinoma tissues between different FIGO stage, histological grade, depth of myometrial invasion, lymphatic vascular infiltration status and lymph node metastasis were statistically significant (all P<0.05). Kaplan-Meier survival analysis showed that the 5-year survival rate and the survival time in the low expression group were higher than those in the high expression group [78.57% vs 37.04%, (70.67±4.94) months vs (42.14±3.65) months], the difference was statistically significant (χ2=12.839, P<0.01). Cox proportional hazards regression model analysis showed that the FIGO stage [HR=2.248 (95%CI: 1.034-4.887)], myometrial invasion depth [HR=3.055 (95%CI: 1.668-5.592)], lymph node metastasis [HR=3.811 (95%CI: 1.786-8.131)] and the expression of HOTTIP [HR=2.649 (95%CI: 1.026-6.842)] were all independent influence factors for the prognosis of patients with endometrial carcinoma. Conclusion: lncRNA HOTTIP is highly expressed in endometrial carcinoma tissues and associated with malignant progression of patients. It is an independent influencing factor for patients’ prognosis.

Long noncoding RNA HOTTIP overexpression: A potential prognostic biomarker in prostate cancer.

HOXA transcript at the distal tip (HOTTIP) is a long noncoding RNA (lncRNA), which is >200 nucleotides in length. HOTTIP expression has been demonstrated to play a crucial oncogenic role in cancer pathogenesis, and is said to be associated with poor human cancer prognosis. In prostate cancer, HOTTIP has been identified as an oncogene, but its clinicopathologic significance remains unclear. Array-based qRT-PCR was used to investigate lncRNA levels in 10 pairs of prostate cancer tissues and non-neoplastic parenchyma. Tissue microarray (TMA) was constructed using a total of 70 surgically resected prostatic adenocarcinoma tissues obtained from the Korea University Anam Hospital from 2009 to 2013. HOTTIP expression was determined by RNA in situ hybridization(ISH) and was correlated with clinicopathologic features. Increased HOTTIP expression was observed in all available prostate cancer tissue specimens compared with that in paired normal tissue. High HOTTIP expression was positively associated with bad clinicopathologic features, including higher pathologic T stage (p < 0.001), presence of extraprostatic extension (p < 0.001), seminal vesicle invasion (p < 0.001), perineural invasion (p < 0.001), and the tumor involvement of resection margin (p = 0.044). In particular, significantly increased HOTTIP expression was observed in specimens from patients in the high or very high-risk group, according to the 2018 National Comprehensive Cancer Network (NCCN) guidelines (p < 0.001). Also, patients with high HOTTIP expression showed poorer overall survival than those with low expression. In conclusion, we analytically validated the poor prognostic significance of HOTTIP overexpression and its association with bad clinicopathologic features, and present HOTTIP as a potential prognostic biomarker in prostate cancer.

Long non-coding RNA HOTTIP enhances IL-6 expression to potentiate immune escape of ovarian cancer cells by upregulating the expression of PD-L1 in neutrophils.

BACKGROUND: Long non-coding RNA (lncRNA) HOXA transcript at the distal tip (HOTTIP), has been demonstrated to be a vital biomarker when evaluating the prognosis of multiple cancers. Nevertheless, the potential function of HOTTIP in ovarian cancer (OC), a prevalent cancer among women worldwide, remains elusive. Hence, the current study aimed to elucidate the functional relevance of HOTTIP in the development of OC. METHODS: Positive expression of PD-L1 and IL-6 was determined using immunohistochemical staining in the collected OC and normal tissues. The correlation of IL-6 and PD-L1 was analyzed using flow cytometry, Western blot analysis as well as Pearson's correlation coefficient. The interaction among HOTTIP, c-jun and IL-6 was investigated with the use of RIP, ChIP and dual luciferase reporter gene assays. Finally, the effects of HOTTIP on T cell proliferation and infiltration were identified through gain- and loss-of-function studies in vitro and in vivo. RESULTS: HOTTIP, IL-6 and PD-L1 were all highly expressed in OC tissues. A positive correlation was observed between IL-6 and PD-L1 and that between HOTTIP and IL-6 in OC tissues. HOTTIP was noted to promote the expression of IL-6 by binding to c-jun, which resulted in a promoted PD-L1 expression in neutrophils and immune escape while inhibiting T cell proliferation as well as tumor immunotherapy. CONCLUSION: Taken together, our study unveiled that HOTTIP could promote the secretion of IL-6, and consequently up-regulate the expression of PD-L1 in neutrophils, thus inhibiting the activity of T cells and ultimately accelerating immune escape of OC cells. Our study provides a potential therapeutic strategy by targeting HOTTIP in OC.