Transfer RNA-derived small RNAs (tsRNAs) have been implicated in tumor progression and immune regulation in recent years. However, the specific role of tRNA halves (tiRNAs), a subclass of tsRNAs, in modulating immunotherapy response remains unexplored. In this study, 5’-tiRNAGly levels were examined in gastric cancer (GC) patients and found to be upregulated, especially in non-responders to anti-PD-1 therapy. Elevated 5’-tiRNAGly levels were also associated with diminished oxoglutarate dehydrogenase-like (OGDHL) expression. Further exploration revealed that 5’-tiRNAGly bound to DLST and promoted OGDHL destabilization, whereas targeted inhibition of 5’-tiRNAGly restored OGDHL stability through succinylation at lysine 910, enhanced tricarboxylic acid (TCA) cycle activity, and reduced glutamine-derived metabolic reprogramming. Additionally, 5’-tiRNAGly was found to decrease the activity of α-ketoglutarate dehydrogenase and inhibit succinylation of histone H3 at lysine 79 (H3K79suc), thereby downregulating PD-L1 transcription and reducing therapeutic responsiveness to PD-1 inhibitors. Conversely, restoration of this epigenetic modification upon 5’-tiRNAGly inhibition facilitated PD-L1 transcription, thereby sensitizing tumors to anti-PD-1 therapy. Our findings indicate that targeting 5’-tiRNAGly may represent a promising strategy to enhance responsiveness to anti-PD-1 therapy in GC patients.

Gastric cancer (GC) has a high incidence in China. There is a closed-loop structure in circRNAs, which is involved in various cellular biological processes such as tumor development. However, there is a lack of research on the function of circRNAs in GC. In this study, we aimed to explore the potential of circXPO1 as a diagnostic biomarker and the role of circXPO1 in the progression of GC. We screened out circXPO1 through circRNA sequencing. Using exonuclease digestion assay, agarose gel electrophoresis (AGE), Sanger sequencing, and gDNA experiment, we proved that circXPO1 contained a cyclic structure. Quantitative real-time fluorescent Polymerase Chain Reaction (qRT-PCR) was used to detect the expression of circXPO1 in plasma and GC tissues. The receiver operating characteristic curve (ROC curve) was established to evaluate the diagnostic efficacy of circXPO1. The role of circXPO1 was assessed in vitro. The binding sites between circXPO1 and miRNAs were predicted by CircBank, Circinteractome, CircAtlas and miRanda databases. CircXPO1 was up-regulated in 67 GC tissues compared with the adjacent normal tissues (p = 0.0002). It was stable and hard to be degraded, which made it an ideal tumor biomarker. Compared with the patients in the normal control group, the expression level of circXPO1 in plasma was higher in GC patients (p < 0.001) and those with benign lesions (p = 0.0031) with statistically significant differences. CircXPO1 was proved to have satisfactory diagnostic efficacy in distinguishing GC patients from healthy donors (AUC = 0.813, 95% CI: 0.749-0.877). Besides, the diagnostic efficacy, sensitivity, and specificity could achieve 0.853, 78% and 86%, respectively, when circXPO1, CEA and CA199 were used together in diagnosis. In addition, in vitro experiments indicated that circXPO1 knockdown significantly weakened the proliferation, invasion and migration of GC cells. It was also predicted that circXPO1 could serve as a sponge of miR-1248 to regulate the progression of GC.

ABSTRACT Background Circular RNA (hsa_circ_0070354), also known as circPTPN13, was demonstrated in our previous study that it can be used as a biomarker for diagnosing non‐small cell lung cancer (NSCLC). However, the mechanism of its role in the occurrence and development of NSCLC has not been investigated. Methods and Results The cell functions of NSCLC cells are evaluated by transfecting with relevant mimics, inhibitors, or plasmids in cell lines. After knocking down hsa_circ_0070354, the expression of miR‐4322 increased, and RAB3B decreased, and the expression changed inversely after overexpression of hsa_circ_0070354. Mechanically, hsa_circ_0070354 acts as a ceRNA to adsorb miR‐4322 and modulate Ras‐Related Protein Rab‐3B (RAB3B) to facilitate the proliferation, migration, invasion, and EMT of NSCLC cells. MiR‐4322 or RAB3B inhibition restored proliferation, migration, and invasion of NSCLC cells after silencing of hsa_circ_0070354. In addition, hsa_circ_0070354 knockdown suppressed tumor growth in vivo. Conclusions Collectively, our study reveals a promotive role of hsa_circ_0070354 in tumor proliferation, invasion, and migration by regulating the miR‐4322/RAB3B axis and highlights that targeting the axis is a promising strategy for the intervention of NSCLC progression.

With the advancement of next-generation sequencing technologies, transfer RNA (tRNA)-derived small RNAs (tsRNAs) have been progressively elucidated in their biogenesis and classification. tsRNAs are derived from precursor tRNAs (pre-tRNAs) or mature tRNAs and are classified into tRNA-derived fragments (tRFs) and tRNA-derived stress-induced RNAs (tiRNAs) based on their specific endonuclease cleavage sites and are functionally involved in diverse biological processes. tsRNAs are widely distributed within cells and are also present in the male reproductive system. Specifically, tRFs are the predominant tsRNA subspecies with established regulatory functions, whereas tiRNAs are less abundant and their roles remain less characterized. Notably, tsRNAs are particularly abundant in mature sperm, serving as vectors of paternal epigenetic inheritance. Multiple studies have shown that environmental factors and dietary modifications can significantly alter tsRNA profiles in the male reproductive system. Moreover, sperm tsRNAs regulate embryonic quality and are closely associated with male fertility. Collectively, these findings reveal the diverse roles of tsRNAs in male reproductive health and their potential as clinical biomarkers.

Radiotherapy resistance in liver cancer is intrinsically linked to nuclear DNA repair pathways, yet directly targeting intranuclear regulators remains hindered by biological barriers. This study pioneered a cytoplasm-tonucleus regulatory strategy by exploiting tripartite motif-containing protein 31 (TRIM31), a cytoplasmic E3 ligase that controlled nuclear c-MYC through the ROS-Wnt/(3-catenin axis, bypassing the need for direct nuclear drug delivery. TRIM31 knockdown triggered a redox storm, which impaired (3-catenin nuclear translocation, thereby suppressing c-MYC-driven DNA repair through enforced G1/S phase arrest. Then, we engineered platelet-biohybrid vectors (sh-TRIM31@PLT) that exploited tumor-homing platelets to improve the efficiency of drug administration. In vivo tracking revealed 1.65-fold greater tumor-specific sh-TRIM31 accumulation compared to free counterparts. When combined with radiotherapy, sh-TRIM31@PLT induced synergistic tumor eradication, ROS-mediated oxidative storm generation, and catastrophic DNA damage amplification, and resulting in 25.91% tumor volume reduction from baseline. The striking divergence was attributable to shTRIM31@PLT-induced G1/S phase arrest and selective elimination of proliferative cancer cell populations. Our findings not only established cytoplasmic-nuclear crosstalk targeting as a viable solution to radiotherapy resistance, but also demonstrated a clinically translatable platform that reengineers cellular trafficking pathways to conquer intratumoral biological barriers, offering a universal blueprint for solid tumor treatment.

Esophageal cancer (EC) is frequently detected at an advanced stage because of the lack of effective noninvasive screening tools, leading to poor prognosis. This study aimed to identify serum transfer RNA-derived small RNA (tsRNA) biomarkers for early detection of EC. Candidate tsRNAs were screened using the tsRFun database, and a quantitative real-time PCR (qRT‒PCR) method for serum 5'tRF-Val was subsequently established and validated. A total of 289 participants were enrolled, including 124 EC patients (71 with early-stage, 53 with advanced-stage disease), 56 individuals with benign lesions, 77 healthy subjects, and 32 postoperative EC patients. Detection performance was evaluated by receiver operating characteristic (ROC) curve analysis, and correlations with clinicopathological parameters were examined. 5'tRF-Val levels were significantly higher in the serum of EC patients than in that of healthy subjects. Its expression correlated positively with aggressive pathological features, including advanced TNM stage and the presence of metastasis. Notably, 5'tRF-Val demonstrated high detection accuracy for early-stage EC, with an AUC of 0.857. Importantly, it effectively distinguished early EC lesions from benign lesions (AUC = 0.824). Furthermore, longitudinal analysis revealed a significant decrease in 5'tRF-Val levels following radical surgery, suggesting its potential for monitoring treatment response. Serum 5'tRF-Val was identified in this study as a novel and promising noninvasive biomarker for the early detection of EC. Owing to its high detection accuracy, association with disease progression, and postoperative dynamic changes, 5'tRF-Val may help to fill the current gap in noninvasive detection of early EC, but this needs to be further verified in multicentre studies.

tsRNAs are a kind of small non-coding single-stranded RNA, which play an important role in many kinds of tumours. Previous studies have identified that tRF-31-U5YKFN8DYDZDD can be used as a novel tumour biomarker for the diagnosis and prognosis of gastric cancer (GC). In this study, we further explored the regulatory effect of tRF-31-U5YKFN8DYDZDD on tumour biological function in GC cells and related regulatory mechanisms. Inhibition of tRF-31-U5YKFN8DYDZDD can inhibit the proliferation, invasion, migration and angiogenesis of GC cells, while overexpression of tRF-31-U5YKFN8DYDZDD has the opposite effect. BMPER was shown to be a direct target of tRF-31-U5YKFN8DYDZDD in GC. Furthermore, the cytological effects of stable overexpression of BMPER were similar to the inhibition of tRF-31-U5YKFN8DYDZDD. And the attenuation of BMPER expression rescued the tRF-mediated promotion of GC cells. WB showed that up-regulation of tRF-31-U5YKFN8DYDZDD could increase the phosphorylation level of ERK and Smad1/5, and promote the expression of epithelial mesenchymal transition (EMT) and matrix metalloproteinases. Animal experiments in vivo show that down-regulation of tRF-31-U5YKFN8DYDZDD can effectively inhibit tumour growth. These data suggest that tRF-31-U5YKFN8DYDZDD is a new tumour-promoting factor and may be a potential new therapeutic target for GC.

Hepatocellular carcinoma (HCC) is one of the most common digestive system malignancies and a leading cause of cancer-related mortality worldwide. Alpha-fetoprotein (AFP), although widely used, shows limited sensitivity in early HCC, leading to missed diagnoses. Novel molecular markers that complement existing biomarkers are urgently needed. tRNA-derived small RNAs (tsRNAs), a newly recognized class of noncoding RNAs generated from transfer RNAs, are abundant in cancer cells and body fluids and have emerging roles in tumourigenesis. This study aimed to identify dysregulated tsRNAs in HCC and evaluate their clinical value as biomarkers. We performed high-throughput sequencing on 3 pairs of HCC tissues and their matched adjacent normal tissues to profile 5’-tiRNA-iMet-CAT, validating its serum levels by quantitative reverse transcription PCR (qRT-PCR). Molecular features and stability were confirmed by electrophoresis, sequencing, and freeze-thaw assays. We assessed clinical correlations, diagnostic potential, and predicted target genes via multiple databases with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis. Serum 5’-tiRNA-iMet-CAT was significantly elevated in HCC patients versus healthy controls. Its high expression correlated with poor differentiation, larger tumour size, and advanced Barcelona Clinic Liver Cancer stage (all P < 0.05). Receiver operating characteristic analysis confirmed strong diagnostic performance, with levels markedly decreasing postoperatively (P < 0.0001). Serum 5’-tiRNA-iMet-CAT is stable, specific, and clinically relevant. It may serve as a promising noninvasive biomarker for HCC diagnosis and evaluating short-term therapeutic response during the early postoperative phase.

Non-coding RNAs (ncRNAs) have important regulatory functions similar to traditional oncogenes or tumor suppressor genes. Our previous research found a novel pan-cancer downexpressed ncRNA, PANC754. However, its function and underlying mechanism remain obscure in colorectal cancer (CRC). In this study, in vitro and in vivo experiments were performed to determine the function of PANC754. Loss and gain of function experiments, molecular docking experiments, and bioinformatic analysis were utilized to visualize its pathway. Co-culture system was leveraged to explore its effect on synergetic immune checkpoint blockage against CRC. Through a series of studies, we found that overexpressed PANC754 significantly inhibited cell viability, migration, and metastasis and induced notable apoptosis in CRC. The mechanical research found that PANC754 was the nuclear-located and its expression was regulated by m6A modification via METTL3 enzyme, which bound with its RBP PSPC1, then interacted with H3K4me1 to chromatin-accessible inhibit immune evasive molecule LGALS7 and led to suppress CRC progress. Furthermore, we confirmed that prominent upregulation of the immune checkpoint inhibitory (ICI) capability of anti-NKG2A, monalizumab when it was combined with PANC754 overexpression. Collectively, our study revealed that PANC754 is a tumor-suppressing ncRNA to form an ncRNA/RBP/histone repression complex with m6A-dependence, which can enhance the immune therapeutics effect of ICI, suggesting a promising therapeutic target. PANC754 is a novel pan-tumor suppressing non-coding RNA which is m6A-dependent and regulated by METTL3 modification enzyme. PANC754 is located at cellular nuclear and interacts with its RNA binding protein (RBP) PSPC1 and chromatin accessible histone H3K4me1, then can enhance immunotherapy capability of ICB anti-NKG2A against colorectal cancer through the immune evasive molecule LGALS7 signaling. Effect is that novel nuclear PANC754/PSPC1/H3K4me1 repression complex down-regulates the level "Don't eat me" signal LGALS7 to improve the immune efficiency of ICB and induce NK or CTL cell to release perforin and cytokine to kill tumor cells. (Created by Figdraw).

Background: Non-coding small RNA, specifically tRNA-derived small RNAs (tsRNAs), are readily detectable in cancer patients, exhibit remarkable stability, and are present in high abundance. They play a significant role in tumor development. However, the clinical significance of serum tsRNAs in hepatocellular carcinoma (HCC) remains poorly understood. In this study, we explored the impact of a novel tsRNA, named tRF-23-R9J89O9N9, in the adjuvant diagnosis, disease monitoring, and prognosis assessment of HCC. Methods: The tRF-23-R9J89O9N9 was identified as the target molecule through screening the The Cancer Genome Atlas(TCGA) database. Its expression levels were measured using qRT-PCR. Various methods, including agarose gel electrophoresis, Sanger sequencing, gradient dilution experiments, room temperature stability tests, and repeated freeze-thaw assessments, were employed to evaluate the performance of tRF-23-R9J89O9N9. The correlation between tRF-23-R9J89O9N9 levels and clinicopathological parameters was analyzed using the chi(2) test. The diagnostic value of tRF-23-R9J89O9N9 in HCC was assessed with ROC curve analysis, while the prognostic value was evaluated using Kaplan-Meier curves. Results: Serum tRF-23-R9J89O9N9 expression levels were significantly elevated in HCC patients, while levels in postoperative patients were restored to those of healthy subjects. Additionally, the expression of tRF-23-R9J89O9N9 related to TNM stage(P = 0.009), lymph node metastasis(P < 0.0001), and degree of differentiation(P < 0.0001). Furthermore, the combination of AFP, PIVKA-II, and CEA greatly improved the diagnostic value for HCC. Serum tRF-23-R9J89O9N9 was also identified as a potential biomarker for dynamic monitoring and prognosis of HCC. Conclusions: tRF-23-R9J89O9N9 may regard as a potential novel biomarker for the adjuvant diagnosis of HCC.

BACKGROUND:Hepatocellular carcinoma (HCC) is a globally prevalent malignancy associated with high morbidity and mortality. Transfer RNA (tRNA)-derived small RNAs (tsRNAs), a class of small non-coding RNAs originating from tRNA, have emerged as potential therapeutic targets in cancers, including HCC. However, the specific tsRNAs involved in HCC and their precise mechanisms remain largely unknown. In this study, we identify and characterize specific tsRNAs involved in the development and progression of HCC, discovering their potential as novel biomarkers for early detection and potential therapeutic targets. AIM:To investigate differentially expressed tsRNAs in HCC, identify potential biomarkers, and elucidate the functions and mechanisms of tsRNAs in HCC. METHODS:Differentially expressed tsRNAs in Barcelona Clinic Liver Cancer 0/A-stage HCC tissues were identified through high-throughput sequencing. Agarose gel electrophoresis, Sanger sequencing, and quantitative polymerase chain reaction were conducted to detect 5'-tRNA halve (tiRNA)-lysine (Lys)-CTT in tissues and serum samples. The diagnostic performance of 5'-tiRNA-Lys-CTT was evaluated using receiver operating characteristic analysis. HCC cell proliferation was examined using the Cell Counting Kit-8 assay, colony formation assay, and 5-ethynyl-2'-deoxyuridine staining. Additionally, the migratory capability of HCC cells was investigated using Transwell assays. RESULTS:The 5'-tiRNA-Lys-CTT demonstrated excellent stability and can be easily detected. Its expression was significantly upregulated in 50 HCC tissues, 110 HCC serum samples, and 5 HCC cell lines vs control groups, and the differences were all significant. This elevated expression was strongly associated with clinicopathological characteristics, particularly tumor size, Barcelona Clinic Liver Cancer stage, and cirrhosis of the liver. Receiver operating characteristic analysis revealed superior detection efficiency of 5'-tiRNA-Lys-CTT exhibits for early-stage HCC compared to established markers. Functional assays revealed that 5'-tiRNA-Lys-CTT overexpression promoted cell proliferation and migration, while its inhibition had the opposite effect. Bioinformatics predictions suggest that 5'-tiRNA-Lys-CTT may influence the development and progression of liver cancer by targeting downstream mRNA via metabolic pathways, cancer pathways, and HCC-specific pathways. CONCLUSION:The 5'-tiRNA-Lys-CTT levels were higher in early HCC patients. 5'-tiRNA-Lys-CTT is a promising diagnostic biomarker for early-stage HCC and may play an oncogenic role in HCC by interacting with downstream mRNA targets via specific pathways.

Gastric cancer (GC) is globally recognized as one of the most widespread malignant tumors. As the symptoms of patients with early GC are ambiguous, the majority of patients are given a diagnosis of advanced GC. Therefore, this necessitates the search for new biomarkers to be utilized in the early diagnosis and screening of GC. Enhancer RNA (eRNA) is a non-coding RNA in transcription by enhancers that is tumor-specific and has a critical function in cancer progression. Our research investigates new eRNAs as bio-diagnostic markers for GC. Four eRNAs with good differential expression in GC were screened by TCGA and University of California, Santa Cruz databases. Quantitative real-time PCR was utilized for testing the level of RASSF8-AS1. The diagnostic effect of RASSF8-AS1 was evaluated using the receiver operating characteristic (ROC). Functional experiments were used to detect the ability of RASSF8-AS1 to affect the metastasis and proliferation in GC cells. The expression of RASSF8-AS1 was obviously elevated in both GC tissues and serum, whereas it was decreased in the serum levels of postoperative GC patients. ROC showed that RASSF8-AS1 was more diagnostically efficient than common diagnostic biomarkers for GC and that diagnostic effectiveness could be better than combining them. The findings of in vitro experiments showed that knocking down the level of RASSF8-AS1 clearly suppressed the ability of growth and metastasis in GC cells. Studies have shown that serum RASSF8-AS1 has the potential to contribute to the progression of GC as a biomarker for diagnosis and prognostic monitoring of GC.

BACKGROUND:Hepatocellular Carcinoma (HCC) is a highly fatal cancer, which is the most common malignant tumor globally. Recently, tRNA-derived small RNA (tsRNA) is a type of non-coding RNA that is closely related to various diseases. However, their precise existence and function in HCC remain unclear. We investigated the role of a novel tsRNA tsRNA-Asp-5-0002 in HCC. METHODS:We screened novel tsRNAs in the TCGA database. There are significant differences in tsRNA-Asp-5-0002 expression in HCC compared with other digestive system tumors. qRT-PCR was applied to confirm the level of tsRNA-Asp-5-0002 in 150 HCC patients, 50 patients with benign liver lesions, and 120 healthy controls. tsRNA-Asp-5-0002 mimics was constructed to transfect HCC cells and explore the effect of tsRNA-Asp-5-0002 on malignant phenotype of HCC. Bioinformatics analysis was used to predict the potential regulatory pathways of tsRNA-Asp-5-0002 in HCC. RESULTS:(1) tsRNA-Asp-5-0002 is down-regulated in HCC. (2) qRT-PCR detection tsRNA-Asp-5-0002 had high sensitivity and specificity. (3) Low-expressed tsRNA-Asp-5-0002 was related to TNM stage, differentiation, and LNM, and the ROC curve indicated that tsRNA-Asp-5-0002 had a better diagnostic performance for HCC. (4) tsRNA-Asp-5-0002 inhibits the proliferation and migration of HCC through the PLCB1/Wnt axis. CONCLUSIONS:tsRNA-Asp-5-0002 can work as an auxiliary biomarker for HCC diagnosis. Over-expression of tsRNA-Asp-5-0002 restrains the malignant process of HCC, which may offer fresh tactics for the adjuvant treatment of HCC.

OBJECTIVES:Gastric cancer (GC) is a global health challenge due to its asymptomatic early stages, low sensitivity of traditional serum biomarkers, and the invasiveness of endoscopy, resulting in delayed diagnosis and poor prognosis. The expression levels of tRNA-derived small RNAs (tsRNAs) significantly influence the emergence and development of malignancies. This study focused on the potential of tsRNAs as serum biomarkers for diagnosing GC. METHODS:Quantitative real-time PCR was used to measure 5'tRF-Lys expression levels, validated by agarose gel electrophoresis and Sanger sequencing. Kaplan-Meier survival curves assessed the association between 5'tRF-Lys expression levels and patient outcomes, while receiver operating characteristic (ROC) curves evaluated diagnostic accuracy. ROC curves showed that 5'tRF-Lys outperformed traditional serum biomarkers, and combining these markers can enhance diagnostic accuracy. RESULTS:The expression level of 5'tRF-Lys effectively differentiated GC patients from gastritis patients and healthy subjects. 5'tRF-Lys expression is negatively correlated with the T stage, lymph node metastasis, tumor-node-metastasis stage, neuro/vascular invasion, and positively correlated with tumor differentiation. Patients with low serum 5'tRF-Lys had a lower survival rate. CONCLUSIONS:5'tRF-Lys shows promise as a sensitive and specific biomarker for GC, including early-stage detection and survival prognosis, offering potential clinical applications.

BACKGROUND:Gastric cancer (GC) remains a major cause of cancer-related mortality globally, largely due to the absence of reliable non-invasive biomarkers for early detection. Circular RNAs (circRNAs), characterized by covalently closed-loop structures, stability, and detectability in circulation, have emerged as promising liquid biopsy candidates. METHODS:circAHSA1 (hsa_circ_0032777) was identified through GEO dataset screening (GSE121445) and validated in GC tissues, serum, and cell lines using qRT-PCR with optimized internal reference selection. Diagnostic performance was assessed using ROC analysis and DeLong tests, evaluating circAHSA1 alone and in combination with CEA, CA199, and CA724. Biological functions were examined through proliferation, apoptosis, migration, and invasion assays. Subcellular localization and potential downstream miRNA interactions were analyzed using nuclear-cytoplasmic fractionation and multi-database bioinformatic prediction. RESULTS:circAHSA1 expression was significantly elevated in GC tissues, serum, and cell lines, and correlated with lymph node metastasis, differentiation status, and TNM stage. Serum circAHSA1 effectively discriminated GC from healthy controls (AUC = 0.787) and gastritis patients (AUC = 0.752), outperforming conventional markers, with statistical superiority confirmed by DeLong analysis. Combined detection further improved diagnostic accuracy (AUC = 0.871). Functionally, silencing circAHSA1 suppressed GC cell proliferation, migration, and invasion while enhancing apoptosis and inducing cell-cycle arrest. Bioinformatic analysis suggested miR-647 and miR-661 as potential downstream targets. CONCLUSIONS:circAHSA1 is a stable, GC-specific circulating biomarker with both diagnostic and functional relevance. These findings support circAHSA1 as a promising candidate for liquid biopsy-based GC detection and a potential therapeutic target.

One of the most common gastrointestinal tumors is gastric cancer (GC), which has a high lethality and a poor prognosis. Recently, it was discovered that mature tRNAs, which are expressed differently in a variety of malignancies, give rise to a novel class of tRNA-derived small RNAs (tsRNAs). In this study, we investigated the role of short RNAs produced from tRNA in GC and possible therapeutic uses. edgeR was used to screen the differentially expressed tsRNAs from the TCGA database and quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) was used to verify the levels of tsRNAs in GC samples. tRF-24-6VR8K09LE9 downregulated in GC was confirmed by detecting serum samples from 114 patients with gastric cancer, 40 patients with gastritis, and 100 healthy controls. The chi-square test displayed that tRF-24-6VR8K09LE9 was highly related to differentiation grade (P = 0.029), T-stage (P = 0.036), lymph node status (P = 0.036), TNM staging (P < 0.0001), and neurological/vascular invasion (P = 0.033). The receiver operating characteristic (ROC) curve indicated that tRF-24-6VR8K09LE9 is more effective than the current diagnostic markers for GC. Furthermore, mechanistic studies verified that tRF-24-6VR8K09LE9 affected the malignant progression of GC through the PI3K/AKT signaling pathway. In conclusion, tRF-24-6VR8K09LE9 can be served as a molecular marker for early GC auxiliary diagnosis. Over-expression of tRF-24-6VR8K09LE9 inhibits the malignant progression of GC, which may provide a new strategy for the adjuvant treatment of GC.

ABSTRACT The bacterium Pseudomonas aeruginosa is able to invade lung epithelial cells and survive intracellularly. During this process, it secretes outer membrane vesicles (OMVs), however, it is currently unclear how OMVs from P. aeruginosa (PA-OMVs) affect lung epithelial cells and their impact on oxidative stress, autophagy, and other physiological activities of lung epithelial cells. In this study, we found that PA-OMVs activated oxidative stress and autophagy in cells. We demonstrated that the NLRP3 (NLR family, pyrin domain containing 3) inhibitor MCC950 can enhance autophagy induced by PA-OMVs. The main function of NLRP3 is related to the body’s immune response and inflammation regulation. MCC950 is the most common inhibitor of NLRP3. Additionally, we showed that PA-OMVs not only enhanced the expression of AMP-activated protein kinase, a key regulator of cellular energy homeostasis, and reactive oxygen species, which play a crucial role in cellular signaling and oxidative stress, but also significantly enhanced the expression of NLRP3. Inhibiting the expression of NLRP3 further enhanced the process of PA-OMVs induced autophagy. These results demonstrate that PA-OMVs activate both autophagy and the NLRP3 inflammasome, with NLRP3 suppressing autophagy to a certain extent, hoping to provide broad ideas for the future applications of PA-OMVs. IMPORTANCE The discovery that lung epithelial cells exposed to outer membrane vesicles from Pseudomonas aeruginosa (PA-OMVs) activate cellular autophagy and induce protective immunity is significant. Specifically, the addition of an NLRP3 inhibitor, MCC950, has been found to decrease NLRP3 targets while simultaneously enhancing the autophagy activity induced by PA-OMVs. This finding unveils a novel theoretical framework for the development of PA-OMVs vaccines, highlighting new targets for enhancing the body’s anti-infective responses. By elucidating the mechanisms through which PA-OMVs trigger autophagy and bolster immune defenses, this research opens avenues for innovative vaccine design strategies aimed at combatting infections effectively.

Since gastric cancer (GC) shows no apparent signs in its early stages, most patients are diagnosed later with a poor prognosis. We therefore seek more sensitive and specific GC biomarkers. Small RNAs formed from tRNAs represent a novel class of non-coding RNAs that are highly abundant in bodily fluids and essential to biological metabolism. This study explores the potential of i-tRF-AsnGTT in gastric cancer diagnostics. To begin with, we sequenced i-tRF-AsnGTT using high-throughput methods. i-tRF-AsnGTT expression levels in GC were determined using real-time fluorescence polymerase chain reaction. Agarose gel electrophoresis, Sanger sequencing, and repeated freezing and thawing were performed to verify molecular properties. A correlation was found between clinical and pathological parameters and i-tRF-AsnGTT expression levels through the χ2 test, and receiver operating characteristic was used to analyze its diagnostic value in GC. In serum, i-tRF-AsnGTT has a low and stable expression level. It can differentiate between patients with gastric cancer and gastritis and healthy donors with better diagnostic efficacy. In combination with clinicopathological parameters, i-tRF-AsnGTT correlates with tumor differentiation; infiltration depth of tumors; tumor, node, metastasis stage; lymph node metastases; and neural/vascular invasion. Serum i-tRF-AsnGTT expression is low in GC patients. Serum from postoperative patients shows increased i-tRF-AsnGTT expression levels. Potentially, this could be used as a biomarker to help diagnose gastric cancer and monitor its prognosis.