Abstract Background The immunosuppressive tumor microenvironment (TME) is a cardinal driver of immune escape and resistance to immunotherapy in renal cell carcinoma (RCC). The N6-methyladenosine (m6A), the most prevalent RNA modification, critically governs tumor aggressiveness and TME reprogramming. We aimed to exploit whether and how tumor-intrinsic m6A modification driven by ZC3H13 (zinc finger CCCH-type containing 13) can dictate the immune landscape of RCC. Methods Loss- and gain-of-function studies were performed in vitro and in allograft tumor model. Tumor-infiltrating immune cells were profiled with flow cytometry and immunostaining. The pivotal cytokine mediated by ZC3H13 depletion was identified through an integrated analysis of RNA-seq and PCR array. The molecular target of ZC3H13 was elucidated through integrated m6A sequencing and RNA sequencing. Results Low ZC3H13 expression was an independent predictor of poor prognosis in RCC. Functionally, ZC3H13 silencing enhanced the malignant behaviors of RCC cells and drove an immunosuppressive microenvironment characterized by aberrant recruitment of myeloid-derived suppressor cells (MDSCs). Mechanistically, ZC3H13 promoted m6A methylation on UNC5CL (UNC-5 Family C-Terminal Like) mRNA, facilitating YTHDC1-mediated stabilization of UNC5CL transcripts, which subsequently suppressed the NF-κB-CSF2 signaling axis, ultimately inhibiting MDSCs accumulation. Furthermore, therapeutic targeting CSF2 in combination with anti-PD1 exerts stronger antitumor effects in RCC by synergistically reversing the MDSCs mediated immunosuppressive microenvironment. Conclusion We identified that ZC3H13 loss unleashes the NF-κB-CSF2 signaling cascade via m6A-dependent silencing of UNC5CL, converting the RCC milieu into an MDSCs-dominated immunosuppressive niche. Targeting CSF2 combined with anti-PD1 may represent a novel therapeutic strategy to achieve better efficacy in RCC.

Immune checkpoint inhibitors (ICIs), particularly PD-1/PD-L1 blockade, represent a cornerstone of treatment for advanced gastric cancer (GC). However, their clinical efficacy is hampered by low response rates and the development of both primary and acquired resistance, underscoring the need for innovative combination therapies. In this study, we investigate the potential of Ubenimex, an immunomodulator and inhibitor of leucyl aminopeptidase 3 (LAP3), in enhancing the therapeutic efficacy of PD-L1 blockade in GC. Using a syngeneic GC mouse model, we demonstrate that Ubenimex significantly augments the efficacy of anti-PD-L1 therapy. We further explore the role of LAP3 in GC progression and find that it is highly expressed in both GC tissues and cells, with elevated LAP3 levels correlating with poor prognosis. Functionally, LAP3 facilitates immune evasion through impaired CD8+ T cell infiltration and cytotoxicity in the GC tumor microenvironment (TME). Notably, our findings reveal that LAP3 enhances PD-L1 expression by binding to UBE3A, an E3 ubiquitin ligase. Ubenimex disrupts the LAP3-UBE3A interaction, leading to restored UBE3A-mediated ubiquitination and degradation of PD-L1. This mechanism reinvigorates CD8+ T cell infiltration and cytotoxic activity within the TME, thereby overcoming resistance to anti-PD-L1 therapy. In conclusion, our study provides a strong rationale for the synergistic potential of Ubenimex in combination with PD-1/PD-L1 blockade, offering a promising strategy to overcome current limitations of ICIs therapy in GC patients.

The long non-coding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR), a tumor suppressor associated with various human cancers, regulates multiple cellular processes. However, the role of HOTAIR in papillary thyroid carcinoma remains unclear. Therefore, the purpose of the present study was to investigate the effect of HOTAIR on the proliferation and migration of PTC cells, and to explore its possible mechanism. The initial step involves conducting an analysis in The Cancer Genome Atlas database of the HOTAIR expression level between PTC and normal tissue samples. Reverse transcription-quantitative PCR was conducted to detect HOTAIR and microRNA (miRNA, miR)-206 expression in each cell line. We assessed cell proliferation using the CCK-8 and MTT assays. We employed the wound-healing and transwell assays to evaluate cell migration. The HOTAIR/miR-206/c-Met targeting relations were verified through dual-luciferase reporter assay. We analyzed changes in protein expression downstream of the c-Met/AKT/mTOR pathway by Western blotting. Our results showed that HOTAIR was highly expressed in PTC cells. Silencing HOTAIR significantly inhibited the proliferation and migration of PTC cells. Conversely, miR-206 expression was downregulated in PTC cells. Importantly, overexpressing miR-206 significantly inhibited PTC cell proliferation and migration. In conclusion, HOTAIR acts as a sponge for miR-206, thereby alleviating its repression of c-Met and activating the AKT/mTOR signaling pathway in PTC cells.

The stimulator of interferon genes (STING) has been an attractive target in cancer immunotherapy. However, natural ligand cyclic dinucleotides (CDNs) and CDN derivatives have demonstrated limited efficacy in clinical trials. This limitation stems from the inherent structure of CDNs, which leads to enzymatic degradation, poor cell internalisation, rapid clearance from the tumour microenvironment, and dose-limiting toxicity. In this study, we developed an amphipathic STING agonist, termed albumin-binding CDNs (AlbiCDNs), to enhance the efficacy of c-di-GMP (CDG) via a lipid-conjugated strategy. The lipid provided a platform for albumin hitchhiking, which facilitated the cytoplasmic delivery of CDG without the use of any exogenous components. In addition, incorporating a stimuli-responsive lipid motif further enhanced the cellular release of CDG. Our results indicated that CDG-1C14, an AlbiCDN, efficiently stimulated the maturation and activation of antigen-presenting cells through STING activation. Furthermore, CDG-1C14 exhibited a significant inhibitory effect on the tumour therapeutic model. Therefore, AlbiCDN is a potent platform for cancer immunotherapy that can expedite clinical translation.

Antibiotic resistome of gut microbiota can be transmitted into the urban ecosystems via fecal pollution, potentially leading to a public health crisis. It is essential to determine the primary sources of fecal contamination and accurately evaluate the health risks for the propagation of antibiotic resistance genes (ARGs) from the intestinal microbiota. In this research, the occurrence, sources and potential hosts of ARGs in urban environmental samples collected from a wastewater treatment plant (WWTP), natural water bodies, tap water and farmland soil were comprehensively investigated. Host-associated fecal indicators for general warm-blooded animals (BacGeneral), humans (crAssphage), and other animals (bovines and swine) were employed for reliable microbial source tracking (MST). Results showed that the fecal indicator BacGeneral was detected in 84.00 % of collected environmental samples, indicating the widespread fecal contamination in local water and farmland. The WWTP was the reservoir and main source of fecal contamination in local environment, harboring the highest total abundances of ARGs (3.85 ± 2.72 ARGs/16S rRNA) and mobile genetic elements (MGEs) (0.32 ± 0.12 MGEs/16S rRNA) from multiple animals and humans. Although the swine-associated indicator was undetected, fecal contamination from both bovines and humans was prevalent in collected samples, with detection rates of pollution indicators at 52.00 % for bovine and 28.00 % for human sources. The co-occurrence of ARGs, fecal indicators and MGEs was analyzed, and significant correlation (P < 0.01) between total ARG abundance and fecal indicator (BacGeneral) in contaminated environments demonstrated that fecal pollution exhibited a great influence on overall resistome in local environment. This research offers a comprehensive understanding of the sources and dissemination of ARGs in feces-polluted urban environments, providing data for the monitoring and prevention of ARG pollution.

KGF, also known as FGF7, is a member of the fibroblast growth factor (FGF) family that binds with high affinity to the FGF receptor 2b (FGFR2b) and regulates various cellular processes, including cell proliferation and differentiation in a variety of tumors. However, its potential role in hypopharyngeal cancer (HPC) remains largely unknown. In our study, we observed increased expression of FGFR2b in HPC. KGF treatment inhibited the expression of the differentiation marker keratin 10 (K10) protein at the post-transcriptional level in FaDu cells. Furthermore, treatment with the proteasome inhibitor MG132 was found to attenuate KGF-induced K10 reduction, suggesting the involvement of the ubiquitin-proteasome system. Using mass spectrometry and immunoprecipitation analysis, we identified the E3 ubiquitin ligase TRIM21 as a K10-interacting protein. Unexpectedly, instead of causing degradation, TRIM21 enhanced K10 protein stability through K6-linked ubiquitination of K10 at lysine 163 (K163) in the context of KGF exposure. Meanwhile, KGF treatment decreased TRIM21 protein levels, which were regulated by the p38 MAPK pathway, leading to K48-linked ubiquitination-mediated degradation of TRIM21. Notably, TRIM21 knockdown significantly promoted proliferation, inhibited differentiation and migration of FaDu cells, whereas TRIM21 overexpression had opposite effects in vitro and suppressed xenograft tumor growth in vivo. Our study demonstrates that TRIM21 may act as a tumor suppressor in HPC. However, TRIM21 overexpression decreased the sensitivity of FaDu cells to 5-fluorouracil, whereas TRIM21 knockdown or KGF administration significantly increased 5-fluorouracil sensitivity. Taken together, these findings highlight the intricate balance between protein stabilization and degradation orchestrated by KGF. This ubiquitination-mediated non-degradation mechanism of TRIM21 may provide novel therapeutic strategies for HPC and other cancers.

Cervical cancer, primarily driven by persistent high-risk human papillomavirus (HPV) infection, remains a global health challenge. While therapeutic vaccines offer promising alternatives to conventional treatments, their clinical efficacy is often hindered by limitations in antigen presentation and the immunosuppressive tumor microenvironment (TME). In this study, we explore the potential of thymoquinone (TQ), a bioactive compound derived from Nigella sativa seeds, to enhance the efficacy of cervical cancer therapeutic vaccines. Utilizing a mouse cervical cancer xenograft model with TC-1 cells expressing HPV16 E6/E7 and ras genes, we observed substantial heterogeneity in vaccine-induced antitumor responses. Mice were stratified into hyporesponsiveness (Hypo) and hyperresponsiveness (Hyper) groups based on tumor progression. The Hyper group exhibited significantly reduced tumor growth, marked by increased CD4+ and CD8+ T cell infiltration, reduced regulatory T cells (Tregs), and altered systemic immune parameters. Metabolite profiling identified 63 differential metabolites, with TQ being notably upregulated in the Hyper group, correlating positively with CD4+ and CD8+ T cells and negatively with Tregs and tumor volume. Combining TQ with the vaccine potentiated antitumor responses, leading to smaller tumors without significant changes in body weight. Immunohistochemical and flow cytometry analyses revealed augmented CD4+ and CD8+ T cell infiltration and reduced Tregs, alongside increased IFN-γ production in the combination of vaccine with (Vax + TQ) group. RNA-Seq analysis of TQ-treated CD8+ T cells revealed upregulation of immune-related genes, including Spp1, Grem1, Cxcl10, Isg15, Lcn2, and Sema3c, highlighting enriched immune pathways. Furthermore, TQ treatment upregulated ISG15 expression and enhanced CD8+ T cell cytotoxicity and cytokine secretion both in vitro and in vivo. In conclusion, our findings suggest that TQ enhances the efficacy of cervical cancer therapeutic vaccines by modulating immune responses, particularly by upregulating ISG15 and boosting CD8+ T cell function. This study provides a theoretical foundation for TQ as a promising adjuvant in cancer immunotherapy.

AIMS:To develop a nano-immunotherapy system combining autophagy inhibition and innate immune activation to reverse the immunosuppressive tumor microenvironment (TME) in pancreatic ductal adenocarcinoma (PDAC). MATERIALS & METHODS:The pH-responsive polymer PC7A was utilized to co-deliver the autophagy inhibitor chloroquine (CQ) and the STING agonist cyclic diguanylate (CDG), forming the CQCP nanosystem. In vitro and in vivo experiments evaluated autophagy inhibition, MHC-I expression, dendritic cell activation, tumor infiltration of lymphocytes, and survival in PDAC-bearing mice. RESULTS:CQCP enhanced MHC-I expression on PDAC cells by 2.1-fold (p < 0.001) and increased activated dendritic cells (CD86+/CD40+) by 3.5-fold (p < 0.01) in the TME. Tumor-infiltrating CD8+ T cells rose by 42.6% (p < 0.001), and systemic immune activation in peripheral lymphoid tissues was observed. CQCP achieved an 86% survival rate in tumor-bearing mice, significantly outperforming monotherapies or free drug combinations. CONCLUSIONS:The CQCP system synergistically reverses PDAC immunosuppression by restoring antigen presentation and activating innate immunity. This dual-targeted strategy demonstrates robust antitumor efficacy and offers a promising immunotherapy approach for PDAC.

Objective To investigate the immunomodulatory effects of CXCL8 on the microenvironment in colorectal cancer (CRC). Methods Subcutaneous transplanted tumor model in BALB/c mice was established using CXCL8-overexpressing CT26, a murine CRC cell line . Tumor growth was monitored, and after three weeks of formation, the tumors were excised, and the spleens were harvested. Firstly, the tumor single-cell suspensions were prepared, and the infiltration of M2 macrophages and CD8+ T cells in the tumor microenvironment were detected by flow cytometry. Additionally, the spleen single-cell suspensions were prepared and CD8+ T cells were sorted. T cells were co-cultured with CXCL8-overexpressing CT26 cells in vitro, and the cytotoxicity assays were performed to evaluate the killing ability of T cells. Results Overexpression of CXCL8 promoted the growth of CRC transplanted tumors. Tumor overexpressing CXCL8 exhibited increased the infiltration of M2 macrophages and decreased the infiltration of CD8+ T cells. However, overexpression of CXCL8 in CRC cells did not affect the cytotoxicity of CD8+ T cells in vitro. Conclusion CXCL8-overexpressing CRC cells promoted the infiltration of M2 macrophages and inhibited CD8+ T cell infiltration to generate an immunosuppressive microenvironment in CRC.

Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cell carcinoma and has a poor prognosis. Despite the impressive advancements in treating ccRCC using immune checkpoint (IC) blockade, such as PD-1/PD-L1 inhibitors, a considerable number of ccRCC patients experience adaptive resistance. Therefore, exploring new targetable ICs will provide additional treatment options for ccRCC patients. We comprehensively analyzed multi-omics data and performed functional experiments, such as pathologic review, bulk transcriptome data, single-cell sequencing data, Western blotting, immunohistochemistry and in vitro/in / in vivo experiments, to explore novel immunotherapeutic targets in ccRCC. It was found that immune-related genes VSIG4, SAA1, CD7, FOXP3, IL21, TNFSF13B, BATF, CD72, MZB1, LTB, CCL25 and KLRK1 were significantly upregulated in ccRCC ( Student's t test and p-value < 0.05; 36 normal and 267 ccRCC tissues in raining cohort; 36 normal and 266 ccRCC tissues in validation cohort) and correlated with the poor prognosis of ccRCC patients ( Wald test and p-value < 0.05 in univariate cox analysis; log-rank test and p-value < 0.05 in Kaplan-Meier method; 267 patients in training cohort and 266 in validation cohort). In particular, we found the novel IC target VSIG4 was specifically expressed in inhibitory immune cells M2-biased tumor-associated macrophages (TAMs), conventional dendritic cell 2 (cDC2) cells, and cycling myeloid cells in ccRCC microenvironment. Moreover, VSIG4 showed a closely relation with resistance of Ipilimumab/Nivolumab immunotherapy in ccRCC. Furthermore, VSIG4 promoted the infiltration of M2 macrophages, Tregs, and cDC2 in ccRCC tissues. VSIG4+ + TAMs and VSIG4+ + cDC2s may be a kind of immune cell subtypes related to immunosuppression. VSIG4 may play similar roles with other IC ligands, as it is highly expressed on the surface of antigen-presenting cells and ccRCC cells to inhibit T cells activity and facilitate immune escape. Targeting IC gene VSIG4 may provide a novel immunotherapeutic strategy to ccRCC patients with resistance to existing targeted therapy options.

Ovarian cancer is a gynecological malignant tumor with the highest mortality rate, and chemotherapy resistance seriously affects patient therapeutic outcomes. It has been shown that the high expression of anti‑apoptotic proteins Bcl‑2 and Bcl‑xL is closely related to ovarian cancer chemotherapy resistance. Therefore, reducing Bcl‑2 and Bcl‑xL expression levels may be essential for reversing drug resistance in ovarian cancer. ABT‑737 is a BH3‑only protein mimetic, which can effectively inhibit the expression of the anti‑apoptotic proteins Bcl‑xL and Bcl‑2. Although it has been shown that ABT‑737 can increase the sensitivity of ovarian cancer cells to cisplatin, the specific molecular mechanism remains unclear and requires further investigation. In the present study, the results revealed that ABT‑737 can significantly increase the activation levels of JNK and ASK1 induced by cisplatin in A2780/DDP cells, which are cisplatin‑resistant ovarian cancer cells. Inhibition of the JNK and ASK1 pathway could significantly reduce cisplatin cytotoxicity increased by ABT‑737 in A2780/DDP cells, while inhibiting the ASK1 pathway could reduce JNK activation. In addition, it was further determined that ABT‑737 could increase reactive oxygen species (ROS) levels in A2780/DDP cells induced by cisplatin. Furthermore, the inhibition of ROS could significantly reduce JNK and ASK1 activation and ABT‑737‑mediated increased cisplatin cytotoxicity in A2780/DDP cells. Overall, the current data identified that activation of the ROS‑ASK1‑JNK signaling axis plays an essential role in the ability of ABT‑737 to increase cisplatin sensitivity in A2780/DDP cells. Therefore, upregulation the ROS‑ASK1‑JNK signaling axis is a potentially novel molecular mechanism by which ABT‑737 can enhance cisplatin sensitivity of ovarian cancer cells. In addition, the present research can also provide new therapeutic strategies and new therapeutic targets for patients with cisplatin‑resistant ovarian cancer with high Bcl‑2/Bcl‑xL expression patterns.

Ferroptosis is a type of cell death that is strongly associated with the cellular redox state. Glutathione is the key to buffering lipid peroxidation in ferroptosis and can also modify proteins by S-glutathionylation under oxidative stress. Here, we showed that the strong associations among glutathione pools, protein S-glutathionylation, and susceptibility to ferroptosis existed broadly in ferroptosis induced by erastin or acetaminophen. Deficiency of CHAC1, a glutathione-degrading enzyme, led to decreased glutathione pools and reduced protein S-glutathionylation, improved liver function and attenuated hepatocyte ferroptosis upon acetaminophen challenge, which could be retarded by CHAC1 overexpression. We conducted quantitative redox proteomics in primary mouse hepatocytes to identify glutathione pool-sensitive S-glutathionylated proteins and found that S-glutathionylation is required to maintain the function of ADP-ribosylation factor 6 (ARF6). Our data suggest that aberrant ARF6 S-glutathionylation increases the labile iron pool by delaying the recycling of transferrin receptors, thereby promoting ferroptosis. Our study reveals the importance of protein S-glutathionylation in conferring cell resistance to ferroptosis. ![Figure][1]</img> HIGHLIGHTS 1. Highly upregulated CHAC1 decreases glutathione pools and protein S-glutathionylation. 2. Reduced protein S-glutathionylation associated with decreased glutathione pools promotes ferroptosis. 3. S-glutathionylation of ARF6 at Cys90 promotes ARF6 activation. 4. Reduced S-glutathionylation of ARF6 provides a labile iron pool to drive ferroptosis. [1]: pending:yes

Abstract Introduction: The inability to predict treatment response results in unnecessary toxicity, decreased efficacy and survival. There is a clinical need for an effective biomarker to select patients for cancer treatment. A promising biomarker for treatment efficacy is response testing on patient-derived organoids (PDOs). For metastatic colorectal cancer (mCRC), larger prospective studies are needed to evaluate the predictive value of standardized PDO screens. Methods: OPTIC is an ongoing prospective multicenter trial (NL61668.041.17, started 2018) that will evaluate the value of tumor PDOs in therapy response prediction. We establish PDOs for mCRC patients from newly obtained biopsies, prior to the start of treatment. We compare PDO sensitivity (area under drug response curve) with patient response measured by changes in biopsied metastatic lesion size on CT scans. Secondary outcome measures are RECIST response and progression-free survival. We evaluate the feasibility of using PDOs as a biomarker by examining drug screen availability in a clinically relevant timeframe of 4 weeks. From 2020 onwards, we incorporated whole genome sequencing (WGS) on fresh-frozen biopsies to correlate drug response in vitro and in vivo, to genetic tumor characteristics. Additionally, PDO screens with experimental drugs based on genetic alterations identified by WGS could be performed. Trial inclusion will continue until 85 PDOs from clinically evaluable patients for standard of care (SOC) treatment are established, with expected completion in Q1 2024. Results: Currently, 206 patients were included in 5 hospitals and 180 biopsies were taken. PDO establishment success gradually increased from 22% in 2018-2020 to 75% in 2023, yielding an overall success rate of 50% (n=84 PDOs and 13 in culture). Critical factors in optimizing culture success were replacement of several growth factors in the culture medium, reducing time to processing and more frequent handling of biopsies. We have established PDOs for comparing with patient response for the following treatment categories: 5-fluorouracil (n=4), CAPOX/FOLFOX (n=23), irinotecan/FOLFIRI (n=11), FOLFOXIRI (n=12), trifluridine/tipiracil (n=14), panitumumab (n=14), encorafenib-cetuximab (n=2), non-SOC drugs (n=4). We have successfully screened the first 8 PDOs in consistent technical triplicates and biological duplicates (R<0.88, p<0.02). WGS succeeded in 76% (n=75), primarily failing when tumor percentage was <20%. Conclusion: We improved PDO establishment success to 75% which is essential to facilitate the implementation of PDOs in clinical treatment. OPTIC will enhance the clinical application of PDOs by defining thresholds for PDO sensitivity and analyzing the diagnostic power for different treatments. To enable personalized treatment in clinical practice, PDO screening should guide mCRC treatment and result in enhanced chance of response and reduced over- and mistreatment. Citation Format: Lidwien P. Smabers, Emerens Wensink, Carla S. Verissimo, Mayke Doorn, Tao Yang, Timo Voskuilen, Yasmine Abouleila, Maarten A. Huismans, Liselot Valkenburg-van Iersel, Geert A. Cirkel, Elske C. Gootjes, Frank J. Jeurissen, Guus M. Bol, Hilde H. Nienhuis, Manon M. Braat, Bas Penning de Vries, Sjoerd G. Elias, Edwin Cuppen, Robert G. Vries, Onno Kranenburg, Miriam Koopman, Sylvia F. Boj, Jeanine M. Roodhart. Organoids to predict treatment response in metastatic colorectal cancer (OPTIC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5174.

Siglec-15 is a highly conserved member of the Siglec family, expressed on osteoclasts, a subset of myeloid cells and some cancer cells. Except for regulating osteoclast differentiation, Siglec-15 engages in immunoregulation as an immune suppressor. Siglec-15 functions as an immunosuppressive molecule in tumor-associated macrophage -mediated T cell immunity in the tumor microenvironment (TME), which makes Siglec-15 to be an emerging and promising target for normalization cancer immunotherapy. Besides, Siglec-15 interacts with sialylated pathogens and modulates host immune response against microbial pathogens by altering cytokine production and/or phagocytosis, which further broadens the underlying pathophysiological roles of Siglec-15. The fact that N-glycosylation and sialylation of Siglec-15 play a pivotal role in Siglec-15 biological function indicates that tar-geting certain post-translational modification may be an effective strategy for targeting Siglec-15 therapy. In-depth exploring Siglec-15 biology function is crucial for better design of Siglec-15-based therapy according to different clinical indications.

目的:探讨阿司匹林(aspirin,又称acetyl salicylic acid,ASA)对人肾癌细胞系ACHN和786-O细胞增殖、迁移、凋亡和自噬的影响.方法:利用CCK-8 法检测不同浓度的ASA及ASA联合氯喹(chloro-quine,CQ)对ACHN和786-O细胞存活率的影响;应用平板克隆实验检测不同浓度ASA对肾癌细胞克隆形成能力的影响;采用Transwell实验和划痕实验检测肾癌细胞迁移能力;通过Annexin V-FITC/PI实验检测肾癌细胞的凋亡情况;Western blot法检测ASA对肾癌细胞凋亡相关蛋白Bax、Bcl-2 及自噬相关蛋白LC3B、Beclin-1 和p62 表达水平的影响.结果:ASA可显著抑制ACHN和 786-O细胞的存活率,随ASA浓度/时间增加细胞存活率显著下降(P＜0.05);与对照组相比,ASA显著抑制肾癌细胞的增殖和迁移(P＜0.05);ASA 处理增加了肾癌细胞凋亡数目,显著抑制了肾癌细胞凋亡相关蛋白Bcl-2 的表达,上调Bax表达,并促进自噬相关蛋白LC3B和Beclin-1 的表达,抑制p62 的表达(P＜0.05).与CQ处理组相比,ASA联合CQ组显著促进了LC3B的表达抑制p62 的表达(P＜0.05),并显著抑制了肾癌细胞的增殖和迁移并促进其凋亡(P＜0.05).结论:ASA可抑制肾癌细胞的增殖和迁移并促进凋亡和自噬.

It is great significance of identifying valuable biomarkers for early diagnosis and prognostic prediction of colorectal cancer (CRC) patients. This study aimed at developing and validating a miRNAs‐based signature as prognostic tool for CRC patients. The miRNA expression profile of 624 CRC samples (613 tumor tissues and 11 normal tissues) was analyzed, and 523 differentially expressed miRNAs (DEmiRNAs) were identified, in which 191 were downregulated and 332 were upregulated. All patients were randomly divided into a training cohort (N = 308) and an internal validation cohort (N = 200). Using the least absolute shrinkage and selection operator (LASSO) and Cox regression model, a prognostic signature of 10 miRNAs (hsa‐miR‐149‐5p, hsa‐miR‐193b‐5p, hsa‐miR‐193a‐3p, hsa‐miR‐3677‐3p, hsa‐miR‐29a‐3p, hsa‐miR‐200c‐5p, hsa‐miR‐200a‐5p, hsa‐miR‐6854‐5p, hsa‐miR‐216a‐5p and hsa‐miR‐891a‐5p) was developed in the training cohort. The risk score was calculated by the product of the expression level and the coefficients of each miRNA. The prognostic value of 10 miRNAs‐based signature for CRC patients was tested and validated. Survival analysis indicated that high‐risk patients (> 1.10) had a worse overall survival (OS) than low‐risk (≤ 1.10) patients (5‐year OS rate for training cohort: 59.3% vs. 78.9%, p < .001; validation cohort: 48.3% vs. 69.3%, p = .011). The miRNA‐based signature was an independent prognostic factor for CRC patients (HR for training cohort:2.476, 95% CI:1.202–5.098, p = .014; HR for validation cohort:2.050, 95% CI:1.087–3.869, p = .027). The AUC values for 3‐year and 5‐year OS prediction were 0.718 and 0.784 in the training cohort, 0.659 and 0.614 in the validation cohort, respectively. The 10 miRNAs‐based signature provided a proper prognostic stratification for CRC patients, and it might be a promising tool for survival prediction.

抗磷脂抗体综合征(antiphospholipid syndrome,APS)是一种以持续存在抗磷脂抗体(antiphospho-lipid antibody,APA)为特征、以动静脉血栓形成和病理妊娠为主要临床表现的自身免疫性疾病[1].

The poor prognosis of immunotherapy in patients with colorectal cancer (CRC) necessitates a comprehensive understanding of the immunosuppressive mechanisms within tumor microenvironment (TME). Undoubtedly, the anti-tumor immune cells play an indispensable role in immune tolerance. Therefore, it is imperative to investigate novel immune-related factors that have the capacity to enhance anti-tumor immunity. Here, we employed bioinformatic analysis using R and Cytoscape to identify the hub gene chemokine (C-X-C motif) ligand 8 (CXCL8), which is overexpressed in CRC, in the malignant progression of CRC. However, its specific role of CXCL8 in CRC immunity remains to be elucidated. For this purpose, we evaluated how tumor-derived CXCL8 promotes M2 macrophage infiltration by in vivo and in vitro, which can be triggered by IL-1β within TME. Mechanistically, CXCL8-induced polarization of M2 macrophages depends on the activation of the STAT3 signaling. Finally, immunohistochemistry and multiplexed immunohistochemistry analysis identified that CXCL8 not only enhances PD-L1+ M2 macrophage infiltration but also attenuates the recruitment of PD-1+ CD8+ T cells in murine CRC models. Together, these findings emphasize the critical role for CXCL8 in promoting M2 macrophage polarization and inhibiting CD8+ T cell infiltration, thereby links CXCL8 to the emergency of immunosuppressive microenvironment facilitating tumor evasion. Overall, these findings may provide novel strategy for CRC immunotherapy.

Background: ALKBH5 belongs to the ALKB family consists of a Fe (II) and a-ketoglutarate-dependent dioxygenase. ALKBH5 directly catalyzes the oxidative demethylation of m6A-methylated adenosine. ALKBH5 involves in tumorigenesis and tumor progression, and is often dysregulated in a wide range of cancers, including colorectal cancer. Emerging evidence indicates that the expression of ALKBH5 is associated with the abundance of infiltrating immune cells in the microenvironment. However, how ALKBH5 affects immune cell infiltration in the microenvironment in colorectal cancer (CRC) has not been reported. The aim of this study was to identify how the expression of ALKBH5 affects the biological behaviors of CRC cell lines and regulates the effects on infiltrating CD8(+) T cells in CRC microenvironment with its specific mechanism.Methods: Firstly, the transcriptional expression profiles of CRC were downloaded from TCGA database and integrated via R software (4.1.2). Between CRC and normal colorectal tissues, ALKBH5 mRNA expressions were compared (Wilcoxon rank-sum). We further identified the expression levels of ALKBH5 in CRC tissues and cell lines through quantitative PCR, western blot, and immunohistochemistry. Then, how ALKBH5 affects the biological behaviors of CRC cells were confirmed by gain- and loss-of-function analysis. Furthermore, the relationship between ALKBH5 level and 22 tumor-infiltrating immune cells was examined through CIBERSORT in R software. Furthermore, we explored the correlation between ALKBH5 expression and tumor-infiltrated CD8(+), CD4(+) and regulatory T cells by utilizing the TIMER database. Finally, the association between chemokines and CD8(+) T cells infiltration in CRC was analyzed using GEPIA online database. qRT-PCR, WB and IHC were used to further determine the effect of ALKBH5 on NF-?B-CCL5 signaling axis and CD8(+) T cells infiltration.Results: Clinically, ALKBH5 expression was downregulated in CRC and low levels of ALKBH5 expression were correlated with poor overall survival (OS). Functionally, overexpression of ALKBH5 reduced the proliferation, migration and invasion of CRC cells, and vice versa. Overexpression of ALKBH5 suppresses NF-?B pathway, thus reduces CCL5 expression and promotes CD8(+) T cells infiltration in CRC microenvironment.Conclusions: ALKBH5 is poorly expressed in CRC, and overexpression of ALKBH5 attenuates CRC malignant progression by inhibiting CRC cell proliferation, migration, invasion and promoting CD8(+) T cells infiltration in the tumor microenvironment through NF-?B-CCL5 axis.

目的 评价Caprini血栓风险评估量表对经外周中心静脉置管(peripherally inserted central catheter,PICC)导管相关性血栓(peripheral inserted central catheters related thrombosis,PICC-CRT)的预测能力.方法 检索Pubmed、Web of Science、Embase、Cochrane Library、万方医学数据库、中国知网、中国生物医学文献数据库、维普中国科技期刊数据库等数据库从建库至 2021 年 12 月 31 日有关Caprini量表评估PICC-CRT的文献,根据纳入研究数据是否存在异质性及异质性的大小决定进行定量或定性综合分析.结果 纳入 4 篇文献共 1010 例患者,均为肿瘤患者,因I2＞75%,异质性大,只进行定性综合.其中 2 篇为前瞻性队列研究,Caprini临界值分别为 6.5 分和 6 分,对应的灵敏度和特异度分别为0.74 和 0.44、0.76 和 0.44;1 篇为病例对照研究,临界值为 7 分,灵敏度和特异度分别为 0.66 和 0.72;1篇未说明研究类型,临界值为 4.5 分,灵敏度和特异度分别为 0.86 和 0.17.结论 Caprini风险评估量表对肿瘤患者的PICC-CRT具有一定的预测能力,但特异度低,总体预测效能较低.