Invasion Ability Research Articles

Extracellular nicotinamide phosphoribosyltransferase visfatin activates JAK2-STAT3 pathway in cancer-associated fibroblasts to promote colorectal cancer metastasis.

Metastasis is one of the major challenges in the treatment of colorectal cancer (CRC), during which cancer-associated fibroblasts (CAFs) in the tumor microenvironment are critically involved. In this study, we aim to explore the regulatory role of extracellular nicotinamide phosphoribosyltransferase Visfatin and its impact on CRC metastasis. To examine the effect of visfatin on CAFs, human CRC tissue-derived CAFs were exposed to visfatin, and the expression of inflammatory factors, activation of JAK-STAT pathway and production of ROS in CAFs were assessed. To examine the effect of visfatin-treated CAFs on CRC metastasis, human CRC cell line SW480 or SW620 were cultured with the conditioned medium derived from visfatin-treated CAFs, and the invasion and migration ability of SW480 or SW620 cells were evaluated by transwell migration and matrigel invasion assays. Our previous study found that visfatin, a secreted form of nicotinamide phosphoribosyltransferase that governs the rate-limiting step of NAD synthesis, promoted CRC metastasis. However, little is known about the effect of visfatin on CAFs. The conditioned medium derived from visfatin- treated CAFs promotes the migratory and invasive capability of CRC cells, and enhance lung metastasis in mouse model. Visfatin treatment stimulated the expression of a couple of inflammatory factors in CAFs, which was mediated by visfatin-induced activation of JAK- STAT pathway and accumulation of ROS. Inhibition of JAK-STAT pathway or neutralization of cellular ROS attenuated visfatin-mediated migration and invasion of CRC cells. The present work highlights a critical role of visfatin in the crosstalk between CRC cells and CAFs, which moonlight as a non-metabolic extracellular signal molecule to hijacks JAK-STAT pathway in CAFs to promote CRC metastasis.

PPARγ alleviates damage to chorionic trophoblast cells induced by high glucose and high lipids through regulation of IGF-1.

Overweight and obesity are the most common high-risk conditions that increase the risk of adverse outcomes during pregnancy, childbirth, and the postpartum period. Dysfunctions in trophoblastic peroxisome proliferator-activated receptor gamma (PPARγ) contribute to a variety of related pregnancy disorders. This study investigated whether PPARγ influences chorionic trophoblast cell damage induced by high glucose (HG) and high lipid (HL) by regulating insulin-like growth factor-1 (IGF-1). Human trophoblast HTR-8/SVneo cells were exposed to HG and HL conditions to simulate damaged trophoblasts during pregnancy in vitro. Cell Counting Kit-8 (CCK-8) was used to assess cell proliferation. The Scratch test was used to test cell migration. Cell invasion ability was assessed by Transwell assay. ELISA was used to assess the inflammatory factor levels. Glucose, lactic acid, and adenosine triphosphate (ATP) levels were measured using biochemical kits. High glucose/HL inhibited the proliferation, migration, and invasion of HTR-8/SVneo cells. High glucose and HL increased tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β, and IL-6 expression while decreasing IL-10 expression. High glucose and HL decreased glucose uptake and ATP levels. High glucose and HL reduced the expressiofns of PPARγ, IGF-1, insulin receptor substrate (IRS) 1, IRS2, GLUT1, and GLUT4. High PPARγ expression promoted cell proliferation, migration, and invasion induced by HG and HL, increased glucose uptake and ATP levels and inhibited inflammation. Low IGF-1 expression inhibited cell proliferation, migration, and invasion under HG and HL conditions, reduced glucose uptake and ATP levels, and increased inflammation. Low IGF-1 expression reversed the effects of PPARγ on HTR-8/SVneo cells under HG and HL conditions. Peroxisome proliferator-activated receptor gamma alleviated HTR-8/SVneo cell damage induced by HG and HL by regulating IGF-1, suggesting a potentially effective approach for treating gestational obesity.

NTRK fusion promotes tumor migration and invasion through epithelial–mesenchymal transition and closely interacts with ECM1 and NOVA1

BackgroundThe NTRK fusion gene is a rare cancer driver and a typical representative "diamond mutation". Its unique role in tumor progression is highly important for the clinical diagnosis and treatment of patients with tumors. We searched for NTRK fusion-positive patients in our hospital. As of August 2022, a total of 8 patients were affected. We discovered that NTRK fusion was associated with enhanced tumor invasion and migration ability. Previous reports also support this finding, but its underlying mechanism has not been elucidated.MethodsWe undertook a comprehensive exploration of the correlations between NTRK fusions and tumor invasion as well as migration by analysing clinical data, performing bioinformatics analysis via public databases, and conducting in vitro cell experiments.ResultsWe ascertained that within the thyroid cancer (THCA) dataset and the pancancer dataset, ECM1 and NOVA1 were coexpressed with NTRKs. Additionally, they demonstrated a significant association with the activity of the epithelial‒mesenchymal transition (EMT) pathway. Furthermore, these genes are overexpressed in various cancers and are associated with advanced clinical stage and increased aggressiveness. Our in vitro study revealed that larolutinib potentially inhibited the invasion and metastasis ability of NTRK-fused cells. Interestingly, contrary to previous findings, the repression of ECM1 increased the migration and invasion ability of NTRK-fused tumor cells.ConclusionsNTRK fusion tumors present heightened migratory and invasive potential in clinical settings. Further experiments confirmed the significant inhibitory effects of TRK inhibitors on the migration and invasion abilities of these cells. There is a complex relationship between ECM1, NOVA1 and NTRK fusion; however, further research is needed to determine whether NTRK fusion promotes tumor metastasis through these two genes.

GXYLT2: an emerging therapeutic target and predictive biomarker for anti-PD-1 efficacy in clear cell renal cell carcinoma.

There are studies reporting that glucoside xylosyltransferase 2 (GXYLT2) has a role in promoting tumor progression, but its role in clear cell renal cell carcinoma (ccRCC) remains unclear. In this study, RT-qPCR and western blotting were employed to detect the expression level of GXYLT2. RNA interference assays were used to knock down GXYLT2. CCK-8, wound healing assays, clone formation assays, and Transwell assays were utilized to investigate the function of GXYLT2. Bioinformatics analysis was used to explore the tumor microenvironment and potential biological mechanisms. We found that the expression level of GXYLT2 in ccRCC was higher than that in adjacent normal renal tissues. Patients with high GXYLT2 expression have worse clinical outcomes. Knockdown of GXYLT2 inhibits the proliferation, invasion, migration, and clone formation ability of ccRCC cells. Enrichment analysis uncovered that GXYLT2 participates in Wnt, cell cycle, and actin cytoskeleton regulation signaling pathways. After receiving anti-PD-1 therapy, patients with high GXYLT2 expression had longer progression-free survival compared with those with low GXYLT2 expression. In conclusion, GXYLT2 is a novel potential therapeutic target for ccRCC. Meanwhile, GXYLT2 can be used as a novel marker for predicting immunotherapeutic response.

Cancer-associated fibroblast-derived MMP11 promotes tumor progression in pancreatic cancer.

Matrix metalloproteinase 11 (MMP11), a zinc-dependent endopeptidase involved in extracellular matrix degradation and remodeling, has been identified as a tumor promoter in multiple cancer types. However, its expression pattern and role in pancreatic ductal adenocarcinoma (PDAC) remain unclear. In this study, elevated MMP11 expression was identified in PDAC tissues and was associated with diminished survival. Integrated single-cell RNA sequencing and co-immunofluorescence staining revealed that MMP11 was predominantly expressed in cancer-associated fibroblasts (CAFs). Mechanistically, cancer cell-derived TGF-β1 mediated CAF activation via the pSmad2/3 pathway and accompanied by MMP11 production. Additionally, MMP11 knockdown in CAFs impaired the proliferative and invasive abilities of AsPC-1 and BxPC-3 cells invitro; which could be rescued by adding recombinant MMP11. Similarly, co-injection of AsPC-1 cells with MMP11-knockdown CAFs into nude mice significantly suppressed tumor growth and liver metastasis compared with tumors bearing unmodified CAFs. Furthermore, we confirmed that CAF-derived MMP11 may drive the epithelial-mesenchymal transition process of PDAC cells to promote tumor invasion via the PI3K/AKT pathway rather than extracellular matrix remodeling. Collectively, we uncovered a crosstalk between cancer cells and CAFs mediated by TGF-β1 and MMP11 that drives the progression of PDAC.

Ailanthone disturbs cross-talk between cancer cells and tumor-associated macrophages via HIF1-α/LINC01956/FUS/β-catenin signaling pathway in glioblastoma

BackgroundAn increasing number of studies have focused on ailanthone (aila) due to its antitumor activity. However, the role of ailanthone in glioblastoma(GBM) has not been investigated before. This study aims to explore the biological function and the underlying mechanism of ailanthone in GBM.MethodsThe microarray analysis was used to screen out down-stream long non-coding RNAs (lncRNAs) targeted by ailanthone. Real-time PCR(RT-PCR) assay was used to examine LINC01956 expression levels. Colony-formation, Methylthiazolyldiphenyl-tetrazolium bromide(MTT), cell-cycle, organoids culture and in-vivo tumorigenesis assays were used to examine cell growth in vitro and in vivo. Boyden assay was used to examine cell invasion ability in vitro. RNA immunoprecipitation and RNA-protein pull-down assays were used to examine the interaction between LINC01956 and FUS protein. Chromatin Immunoprecipitation(ChIP) assay was used to examine HIF1-α-binding sites in the LINC01956 promoter.ResultsAilanthone decreased GBM cell growth in vitro and in vivo via inducing ferroptosis. Ailanthone treatment exhibited blood‒brain barrier(BBB) permeability and specifically targeted the tumor area. LINC01956 was identified as a down-stream target of Ailanthone. LINC01956 exerted as an onco-lncRNA in GBM. M2 polarization of macrophages induced by exosomes derived from glioma cells overexpressing LINC01956 accelerated GBM progression. Mechanistically, we found that LINC01956 bound to FUS and reduced its ubiquitination. LINC01956 evoked nuclear translocation of phosphorylated (p)-β-catenin by recruiting FUS. Furthermore, under hypoxic conditions, LINC01956 was regulated by HIF-1α. Ailanthone decreased the expression of LINC01956 via suppressing HIF-1α.ConclusionTaken together, our data revealed for the first time that ailanthone regulated HIF-1α/LINC01956/FUS/β-catenin signaling pathway and thereby inhibited GBM progression.

Nrf2 depletion enhanced curcumin therapy effect in gastric cancer by inducing the excessive accumulation of ROS

Gastric cancer (GC) is the most common malignant tumor of the gastrointestinal tract and currently has a poor clinical outcome. Turmeric’s rhizome contains a polyphenolic component called curcumin (Cur), which has been demonstrated to inhibit a variety of tumor cells, such as pancreatic, colon, lung and gastric cancers. However, it remains to be elucidated how Cur functions in GC and what molecular processes underlie it. Here, Cur showed a stronger inhibitory effect on GC cells AGS and HGC27. In addition, Cur’s inhibition of GC cells growth was accompanied by increased ROS production, triggering of the Keap1-Nrf2 signaling pathway, and increased transcription of its downstream antioxidant genes HO-1, GCLM, and NQO1. However, when a ROS scavenger NAC was used, the inhibitory effect of Cur on GC cells was reversed. Nuclear factor erythroid 2-related factor 2 (Nrf2) is overexpressed or activated in cancers to shield cancer cells from oxidative damage by responding to oxidative stress (OS). Cur has been found to act as an activator of Nrf2. Notably, compared with Nrf2 knockdown and Cur alone, the combination of the two dramatically increased Cur-induced ROS overaccumulation and inhibition of GC cells proliferation, migration, and invasive abilities. Consistent with in vitro experiments, Cur combined with Nrf2 knockdown significantly inhibited tumor growth in nude mice transplanted with AGS cells. Therefore, we concluded that Nrf2 depletion enhanced Cur therapy effect in GC by inducing the excessive accumulation of ROS, indicating that this is a promising treatment strategy.

SDC1 Influences Keloid Fibroblasts Migration and Invasion via Targeting Wnt/β-Catenin Signaling Pathway Mediated EMT.

Keloid is the result of abnormal wound healing, puzzled by the invasive growth and high recurrence rate attributed to its complex pathogenic mechanism. Syndecan1 (SDC1) contributes to regulating cell migration and invasion by activating epithelial-mesenchymal transition (EMT) in tumor and fibrotic disease. Herein, using western blot analysis, the authors assessed the role of SDC1 on EMT in keloid and its underlying mechanism. The authors found keloid fibroblasts exhibited a higher proportion of mesenchymal phenotypes, and SDC1 was significantly upregulated in keloid fibroblasts. Then, the authors transfected small interfering RNA targeting SDC1 in keloid fibroblasts and tested the abilities of cell migration and invasion, as well as the expression of EMT-related markers, including N-cadherin, vimentin, and E-cadherin. The results showed that the knockdown of SDC1 markedly suppressed the migration and invasion abilities of keloid fibroblasts and reduced the phenotypes of EMT by inhibiting Wnt/β-catenin signaling pathway. The authors' findings suggest that SDC1 may influences keloid fibroblasts migration and invasion through targeting Wnt/β-catenin signaling pathway mediated EMT, which supports its potential value as a therapeutic target for the treatment of keloid.

ZNF503-AS2 is a promising therapeutic target and is associated with the immune microenvironment in glioma.

Glioma is the most common intracranial malignancy, and the available treatment options are poor. Long noncoding RNAs (lncRNAs) have been reported to be involved in the malignant progression of glioma. The role of ZNF503-AS2 in glioma has not been reported. We screened ZNF503-AS2 with upregulated expression in glioblastoma (GBM) by analyzing the TCGA, CGGA and GTEx databases. Single sample gene set enrichment analysis (ssGSEA) was used to calculate the enrichment of immune cells and signaling pathways in glioma samples. Single-cell datasets were used to analyze the distribution of ZNF503-AS2. In vitro experiments were used to investigate the biological function of ZNF503-AS2. ZNF503-AS2 was highly expressed in glioma and was associated with poor prognosis, malignant progression and infiltration of immunosuppressive cells. Single-cell transcriptomic analysis showed that ZNF503-AS2 was mainly expressed in macrophages and tumor cells. Further analysis revealed that immunotherapy may have better efficacy in patients with low ZNF503-AS2 expression. In vitro experiments showed that knockdown of ZNF503-AS2 reduced the proliferation, invasion and migration ability of glioma cells, induced G2/M cell cycle arrest and promoted apoptosis. ZNF503-AS2 might be a valuable biomarker for predicting the prognosis of glioma patients and a potential target for glioma therapy.

Study on miR-29a inhibiting papillary thyroid carcinoma by downregulating TAGLN2

ObjectiveThis study aims to explore the possible mechanism of miR-29a inhibiting papillary thyroid carcinoma (PTC) through the downregulation of TAGLN2. The paper expects to find novel targets for PTC treatment and prognosis improvement, and is also of great significance in enhancing the overall therapeutic level of PTC and improving life quality of patients. MethodsChanges in PTC cell growth, migration, and invasive ability were respectively detected using MTS assays, wound healing experiments, and Matrigel matrix invasion assays. Expressions of E-cadherin, N-cadherin, vimentin, and MMP2 were determined by Western blotting. Differences in protein expressions between groups were compared. Overexpression and knockdown of TAGLN2 were performed in K1 cells, and the expression levels of PI3K/AKT pathway protein in K1 cells were detected by Western blotting. ResultmiR-29a significantly inhibited the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of K1 cells. Overexpression of miR-29a played an important role in improving the malignant phenotype of PTC by regulating the expression of actin-binding protein TAGLN2. Overexpression of TAGLN2 can counteract the inhibitory effect of miR-29a on K1 cells. TAGLN2 can activate the PI3K/AKT signaling pathway, indicating that it may promote the progress of EMT by activating PI3K/AKT. ConclusionThe miR-29a/TAGLN2 axis may affect the malignant phenotype of K1 cells by regulating downstream PI3K/AKT signaling pathways, thus providing a new target for the treatment of PTC.

ADAM17 promotes colorectal cancer migration and invasion by regulating the TGF-β/Smad signaling pathway

ObjectiveInvestigate ADAM17 expression in colorectal cancer (CRC) at molecular and cellular levels and its potential mechanism in promoting tumorigenesis by regulating CRC cell migration and invasion. Materials and methodsThe study measured ADAM17 mRNA and protein levels in colorectal cancer cells and tissues using qPCR and immunohistochemical staining, and assessed the cells' proliferation, migration, and invasion abilities. ResultsADAM17 expression was significantly higher in CRC tissues than in non-cancerous tissues and was linked to metastasis and poor prognosis in CRC patients. Silencing ADAM17 reduced cell migration and invasion. Mechanistically, knocking down ADAM17 decreased the expression of TGF-β/Smad pathway-related proteins, which inhibited proteins associated with migration and invasion, thus impairing these cellular processes. ConclusionADAM17 likely promotes the migration and invasion of CRC cells by regulating the TGF-β/Smad signaling pathway. This study aids in understanding the molecular mechanisms of CRC metastasis and development, and supports the development of new therapeutic targets.

Upregulation of M6A Reader HNRNPA2B1 Associated with Poor Prognosis and Tumor Progression in Lung Adenocarcinoma.

Lung cancer is the most prevalent malignancy worldwide, and lung adenocarcinoma (LUAD) accounts for a substantial proportion of all cases. N6-methyladenosine (m6A) is the most frequent post-transcriptional modification in mRNAs that also plays a role in cancer development. Heterogeneous nuclear ribonucleoprotein A2/B1 (HNRNPA2B1) is a reader of m6A modification, which can affect tumor invasion, migration, and proliferation. The purpose of this study was to explore the prognostic factors of LUAD based on m6A through bioinformatics analysis. The expression levels and prognostic significance of HNRNPA2B1 in LUAD were analyzed on the basis of data extracted from the UALCAN, GEPIA, NCBI-GEO, Human Protein Atlas, STRING, miRDB, TargetScan, PROMO, Starbase, UCSC Xena browser, TIMER, and TISIDB databases. HNRNPA2B1 protein and mRNA levels in several LUAD cell lines were detected by western blotting and qRT-PCR. CCK8, wound-healing and transwell assays were performed to evaluate the proliferation, invasion, and migration abilities of LUAD cells. HNRNPA2B1 mRNA was found to be significantly overexpressed in LUAD tissues, and its high levels correlated with poor OS and DFS. The genes co-expressed with HNRNPA2B1 were related to mRNA production, cell cycle, and histone binding. To determine the mechanistic basis of HNRNPA2B1 in LUAD, we next predicted the microRNAs and transcription factors that were directly associated with HNRNPA2B1, as well as copy number changes. In addition, it was found that HNRNPA2B1 expression was significantly related to CD4+ T cells, neutrophils, lymphocytes, immunomodulators, and chemokines. Besides, knocking down HNRNPA2B1 in the LUAD cells led to a significant reduction in their proliferation, invasion, and migration rates in vitro. Elevated HNRNPA2B1 is a risk factor in LUAD and portends a poor prognosis.

Blocking CXCR3B Expression Increases Tumor Aggressiveness in Hepatocellular Carcinoma.

CXCR3B has been positively involved in the inhibition of cancer and angiogenesis. The present study investigated the role of CXCR3B in a cell model of hepatocellular carcinoma, SK-Hep1. The blockade of CXCR3B expression in SK-Hep1 was investigated in terms of cell viability, cell cycle, and cell apoptosis using MTT assay and flow cytometry. In addition, the effect of blocking CXCR3B expression on cell migration and invasion was examined using scratch motility, transwell migration, and invasion assays. Furthermore, the cytotoxic effect of NK-92 cells against CXCR3B blocked SK-Hep1 was analyzed using the CytoTox96 assay, and the expression of NKp30+, NKG2D+, and NKG2C+ on NK-92 cells in a co-culture with SK-Hep1 was measured using flow cytometry. Blocking CXCR3B expression had no effect on the viability, cell cycle or apoptosis of SK-Hep1 cells. However, blockade of CXCR3B expression significantly increased the migratory and invasive ability of SK-Hep1 along with increased protein expression of slug, vimentin, and N-cadherin. CXCR3B blockade reduced the cytotoxicity of NK-92 against SK-Hep1 and inhibited the expression of activating receptors, NKp30+, NKG2D+, and NKG2C+ in NK-92 cells. CXCR3B may play a positive role in suppressing HCC by attenuating natural killer cell cytotoxicity against HCC.

Toxic effects of dibutyl phthalate on trophoblast through mitochondria mediated cellular dysfunction.

Dibutyl phthalate is a chemical commonly used as a plasticizer in the production of daily necessaries, such as cosmetics and toys. Although several toxic effects of dibutyl phthalate have been confirmed, those related to pregnancy are unknown. Trophoblasts are critical for fetal and placental development, and trophoblast damage may cause preeclampsia. This study aimed to confirm the toxic effect of dibutyl phthalate on trophoblasts. We used the human trophoblast cell line HTR-8/SVneo and human choriocarcinoma JEG-3 cells as a placental trophoblast model to investigate the toxic effects of dibutyl phthalate. Both cell lines were treated with dibutyl phthalate (0-20 μg/mL) to verify the mechanisms regulating trophoblast function. Dibutyl phthalate treatment significantly reduced trophoblast viability, reduced invasion ability, and induced mitochondrial depolarization. Ultimately, dibutyl phthalate regulated the PI3K and MAPK signaling pathways and the expression of autophagy-related proteins ATG5, LC3B, and SQSTM1/p62. We concluded that dibutyl phthalate induced autophagy and effectively weakened trophoblast function. Additionally, we conducted experiments to assess the potential effects of monobutyl phthalate, a metabolite of dibutyl phthalate, on cellular mobility, penetration, and autophagy induction. Our results demonstrated that monobutyl phthalate impaired these functions and weakened the trophoblast barrier, after dibutyl phthalate metabolized. Thus, exposure to dibutyl phthalate and its metabolite monobutyl phthalate can damage trophoblast function, highlighting their potential as hazardous substances that impair trophoblast barrier integrity.

METTL3/YTHDF1 stabilizes CORO6 expression promoting osteosarcoma progression through glycolysis

This study investigates the role of CORO6 (Coronin 6) in the development of osteosarcoma. Osteosarcoma is a common malignant bone tumor in children and adolescents, characterized by rapid and irregular bone growth and a high risk of distant lung metastasis. CORO6 is a member of the Coronin family, known for its conserved WD40 repeat domain. This structure allows CORO6 to inhibit actin dynamics through interactions with F-actin and Arp2/3, thereby affecting the organization of the cytoskeleton. Our research found that in osteosarcoma patients, the levels of CORO6 are significantly elevated. Experimental observations showed that reducing the expression of CORO6 significantly inhibits the growth, migration, and invasion abilities of osteosarcoma cells. Moreover, in vivo experiments demonstrated that the absence of CORO6 effectively inhibits the growth of osteosarcoma in animal models. We also discovered that CORO6 promotes the proliferation, migration and invasion capabilities of osteosarcoma cells by activating the Wnt/β-catenin signaling pathway. Moreover, CORO6 play a critical important role in glycosis of osteosarcoma cells. Mechanically, we found that METTL3/YTHDF1 induced m6A modification of CORO6 mRNA promoted the expression of CORO6 by enhancing its stability. These findings offer new directions for the treatment of osteosarcoma, suggesting that CORO6 could be a novel prognostic biomarker and an effective therapeutic target for patients.In summary, CORO6, as an oncogene, plays a key role in the development of osteosarcoma, providing a crucial theoretical basis for the development of new osteosarcoma treatment strategies.

Curcumin alleviates the aggressiveness of breast cancer through inhibiting cell adhesion mediated by TEAD4-fibronectin axis

Breast cancer (BC) ranks first among women in the world and metastasis is the main reason of cancer death. TEA domain transcription factor 4 (TEAD4) plays an important role in TEAD family members in the Hippo signaling pathway, and its pro-cancer effect is gradually being discovered, but little is known about the mechanism of TEAD4 regulating tumor adhesion and metastasis. Curcumin is an active polyphenol compound with anti-inflammatory and anti-tumor properties. However, its effects and the underlying mechanisms on BC metastasis remain unknown. Here, we show that curcumin reduced the abilities of migration and invasion of BC cells as well as lung metastasis of BC in nude mice by TEAD4 induced. TEAD4 could regulate the transcription level of adhesion molecule Fibronectin (FN1), which is an important component of extracellular matrix participating in tumor cell adhesion and migration processes, and the binding of TEAD4 to the FN1 promoter was suppressed by curcumin. Furthermore, the metastatic mobility was significantly reduced in FN1 knockout BC cells, and FN1 knockout can reverse metastatic potential of TEAD4 overexpressed cells. Our work demonstrates that TEAD4 could promote cell adhesion and migration by binding with FN1 promoter and suggested that TEAD4-FN1 axis in tumor microenvironment is expected to become a potential target for alleviating metastasis of BC of curcumin.Abbreviations: Breast cancer (BC); Bicinchoninic acid (BCA); Bovine Serum Albumin (BSA); Curcumin at 20 μM (Cur20) and 30 μM (Cur30); Dimethyl −sulfoxide (DMSO); Dulbecco’s Modified Eagle’s Medium (DMEM); Extracellular Matrix (ECM); Fetal bovine serum (FBS); Fibronectin (FN1); Immunohistochemical (IHC); Minimum Essential Medium (MEM); The negative control with infected blank lentivirus (NC); Phenylmethanesulfonyl fluoride (PMSF); Polyvinylidene difluoride (PVDF); Real-time quantitative reverse transcription (RT-qPCR); single guide RNA (sgRNA); short hairpin RNA (shRNA); Human TEAD4 shRNA (shTEAD4); Transcriptional coactivator with PDZ-binding motif (TAZ); TEA domain transcription factor 4 (TEAD4); TEAD4 overexpressed (TEAD4-OE); Vascular endothelial growth factor (VEGF); Yes-associated protein (YAP);

Deciphering potential molecular mechanisms in clear cell renal cell carcinoma based on the ubiquitin-conjugating enzyme E2 related genes: Identifying UBE2C correlates to infiltration of regulatory T cells.

Renal clear cell carcinoma (ccRCC) is a highly aggressive and common form of kidney cancer, with limited treatment options for advanced stages. Recent studies have highlighted the importance of the ubiquitin-proteasome system in tumor progression, particularly the role of ubiquitin-conjugating enzyme E2 (UBE2) family members. However, the prognostic significance of UBE2-related genes (UBE2RGs) in ccRCC remains unclear. In this study, bulk RNA-sequencing and single-cell RNA-sequencing data from ccRCC patients were retrieved from the Cancer Genome Atlas and Gene Expression Omnibus databases. Differential expression analysis was performed to identify UBE2RGs associated with ccRCC. A combination of 10 machine learning methods was applied to develop an optimal prognostic model, and its predictive performance was evaluated using area under the curve (AUC) values for 1-, 3-, and 5-year overall survival (OS) in both training and validation cohorts. Functional enrichment analyses of gene ontology and Kyoto Encyclopedia of Genes and Genomes were conducted to explore the biological pathways involved. Correlation analysis was conducted to investigate the association between the risk score and tumor mutational burden (TMB) and immune cell infiltration. Immunotherapy and chemotherapy sensitivity were assessed by immunophenoscore and tumor immune, dysfunction, and exclusion scores to identify potential predictive significance. In vitro, knockdown of the key gene UBE2C in 786-O cells by specific small interfering RNA to validate its impact on apoptosis, migration, cell cycle, migration, invasion of tumor cells, and induction of regulatory T cells (Tregs). Analysis of sc-RNA revealed that UBE2 activity was significantly upregulated in malignant cells, suggesting its role in tumor progression. A three-gene prognostic model comprising UBE2C, UBE2D3, and UBE2T was constructed by Lasoo Cox regression and demonstrated robust predictive accuracy, with AUC values of 0.745, 0.766, and 0.771 for 1-, 3-, and 5-year survival, respectively. The model was validated as an independent prognostic factor in ccRCC. Patients in the high-risk group had a worse prognosis, higher TMB scores, and low responsiveness to immunotherapy. Additionally, immune infiltration and chemotherapy sensitivity analyses revealed that UBE2RGs are associated with various immune cells and drugs, suggesting that UBE2RGs could be a potential therapeutic target for ccRCC. In vitro experiments confirmed that the reduction of UBE2C led to an increase in apoptosis rate, as well as a decrease in tumor cell invasion and metastasis abilities. Additionally, si-UBE2C cells reduced the release of the cytokine Transforming Growth Factor-beta 1 (TGF-β1), leading to a decreased ratio of Tregs in the co-culture system. This study presents a novel three-gene prognostic model based on UBE2RGs that demonstrates significant predictive value for OS, immunotherapy, and chemotherapy in ccRCC patients. The findings underscore the potential of UBE2 family members as biomarkers and therapeutic targets in ccRCC, warranting further investigation in prospective clinical trials.

Long Non-Coding RNA SNHG3 Promotes the Progression of Cholangiocarcinoma by Regulating the miR-151a-3p/STAT5a Axis.

Studies have shown the significance of long non-coding RNAs (lncRNAs) in the development of malignant tumors, including cholangiocarcinoma (CCA). However, the molecular mechanisms through which the lncRNA SNHG3 contributes to CCA development remain unknown. Therefore, the purpose of this work was to investigate SNHG3's role and possible processes in CCA. CCK-8, TUNEL, wound healing, and transwell assays were performed to evaluate the viability, apoptosis, migration, and invasion of CCA cells, respectively. Dual-luciferase reporter and RNA pull-down assays were conducted to verify the relationship between SNHG3 and miR-151a-3p and that between STAT5a and miR-151a-3p. SNHG3 and STAT5a were considerably upregulated and miR-151a-3p was downregulated in CCA tissues and cells. SNHG3 knockdown suppressed the proliferation, apoptosis, migration, and invasive ability of HUCC-T1 cells. Mechanistically, SNHG3 directly targeted miR-151a-3p to promote the development of CCA. Treatment with a miR-151a-3p inhibitor reversed the effects of SNHG3 knockdown on the aggressive behavior of HUCC-T1 cells. Furthermore, STAT5a knockdown counteracted the effects of inhibition of SNHG3 and miR-151a-3p on the aggressive behavior of CAA. SNHG3 promotes CCA progression via the miR-151a-3p/STAT5a axis, providing novel insights into the clinical treatment of CCA.

β-elemene Ameliorates Cisplatin Resistance of Gastric Cancer via Regulating Exosomal METTL3-m6A-ARF6 Axis.

The medial overall survival is low in patients with gastric cancer (GC) at advanced stage, in which drug resistance plays an important role. β-elemene has been established as the suppressed role on GC cell proliferation, however, the concrete mechanism of it remains unclear in cisplatin (DDP)-resistance GC. Cell counting kit-8 (CCK8) assay was used to measure the half maximal inhibitory concentration (IC50) values of DDP in DDP-resistance GC cell lines. Cell apoptotic rates and invasive ability were tested by flow cytometry and transwell assay. Western blot and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) were utilized to detect the protein and mRNA levels of methyltransferase like-3 (METTL3) and ADP ribosylation factor 6 (ARF6). SRAMP websites and methylated RNA immunoprecipitation (MeRIP) assay were applied to predicted m6A sites and verified m6A levels of ARF6 respectively. RNA immunoprecipitation (RIP) was used to explore the interaction between these two molecules. Xenograft tumor models were constructed to demonstrate the effects of β-elemene in vivo. β-elemene improved drug sensitivity and curbed malignant cell activities of DDP-resistance GC cells in vitro. ARF6 was upregulated in DDP-resistance GC cells and tissues, and its overexpression could abrogate the effects on DDP-resistant GC cells mediated by β-elemene treatment. Intracellular and exosomal METLL3 expression were elevated in and from DDP-resistance GC cell lines. Exosomal METTL3 released from DDP-resistance GC cells could counteract the effects of β-elemene on DDP-resistance GC cells partly via regulating ARF6 expression in the m6A-dependent manner. β-elemene could suppress DDP-resistance tumor growth in vivo. In conclusion, β-elemene could repress tumor growth and drug resistance via exosomal METTL3-m6A-ARF6 axis.

Identification of Antisense RNA NRAS-AS and Its Preliminary Exploration of the Anticancer Regulatory Mechanism

Objective: To explore the influence of NRAS-AS on the proliferation, apoptosis, cell cycle, migration, and invasion ability of HCC cells, as well as its underlying mechanisms. Methods: A double-stranded cDNA library for liver cancer cells was constructed, and identified NRAS-AS through High-throughput sequencing, bioinformatics, chain-specific fluorescent quantitative PCR, and RACE. NRAS-AS′s effects on HepG2 and SMMC-7721 cells and gene expression were evaluated. Additionally, the study analyzed the influence of NRAS-AS overexpression on tumor formation in nude mice. Immunohistochemistry and Western blotting were used to detect NRAS protein levels in clinical samples. RT-qPCR examined NRAS-AS and NRAS gene expression in HCC and adjacent tissues. Results: NRAS-AS overexpression suppresses HCC cell proliferation and invasion, induces cell cycle alterations in HepG2 and SMMC-7721 cells, and enhances apoptosis. NRAS-AS interference promoted liver cancer invasion, inhibited apoptosis, and influences the cell cycle. Nude mice overexpressing NRAS-AS showed smaller tumors. NRAS-AS expression in liver cancer patients correlated with clinical factors. RT-qPCR revealed an inverse correlation between NRAS-AS and NRAS gene expression in liver cancer and adjacent tissues. IHC analysis revealed reduced NRAS protein expression in HepG2 and SMMC-7721 cells following NRAS-AS overexpression. The impact of AZA treatment on antisense NRAS-AS and sense NRAS gene expression in liver cancer cells was observed, and antisense. Conclusion: Reduced NRAS-AS expression is frequently observed in HCC and is inversely related to NRAS gene expression, suggesting a role in HCC pathogenesis through NRAS regulation. Targeting antisense RNA NRAS-AS could hold promise as a therapeutic target and diagnostic biomarker for HCC.