Reticulophagy, a selective macroautophagy/autophagy process targeting endoplasmic reticulum fragments via receptors, plays a critical role in cellular homeostasis. This study reveals that CCPG1, a reticulophagy receptor, drives bladder cancer (BLCA) tumorigenesis and confers cisplatin resistance. We observed elevated reticulophagy activity in BLCA cells compared to normal counterparts, particularly under conditions of nutrient stress. CCPG1 expression was significantly upregulated in BLCA patient samples and correlated with poor prognosis. Functional studies demonstrated that CCPG1 knockdown suppressed reticulophagy, leading to decreased cell proliferation and increased apoptosis. Conversely, overexpression of the wild-type CCPG1, but not a MAP1LC3/LC3-binding-deficient variant, rescued reticulophagy and promoted tumor growth. Notably, we found that cisplatin treatment inhibited reticulophagy by downregulating CCPG1 expression through the ATM-CHEK2/Chk2 signaling pathway. CCPG1 knockdown synergistically enhanced cisplatin cytotoxicity to BLCA cells, while CCPG1 overexpression conferred resistance. These findings highlight CCPG1-mediated reticulophagy as a driver of BLCA progression and as a potential prognostic biomarker and therapeutic target.

Glioma is the most common primary brain tumor, and targeting glioma stem cells (GSCs) has become a key aspect of glioma treatment. In this study, we discovered a molecular network in which circRNA forms an R-loop structure with its parental gene to regulate the biological behavior of GSCs. Genes with abnormal expression in GSCs were screened using RNA-seq and circRNA microarray analyses. The study results showed that high expression of YTHDC1 in GSCs promoted the transportation of N6-methyladenosine (m6A)-modified circPOLR2B from the nucleus to the cytoplasm. Decreased circPOLR2B levels in the nucleus resulted in fewer R-loop structures formed with its parental gene POLR2B. This reduction in R-loop structures relieved the inhibitory effect on POLR2B transcription and upregulated PBX1 expression through alternative polyadenylation (APA) action, thereby promoting the malignant biological behavior of GSCs. Knockdown of YTHDC1, POLR2B, and PBX1 reduced xenograft tumor volume and prolonged the survival of nude mice. The YTHDC1/circPOLR2B/POLR2B/PBX1 axis plays a regulatory role in the biological behavior of GSCs, offering potential targets and novel strategies for the treatment of glioma.

BackgroundCircular RNAs (circRNAs), which are covalently closed non-coding RNAs, are frequently dysregulated in cancer. However, their precise role in bladder cancer (BCa) remains largely unknown.MethodsExpression of hsa_circ_0005320 in tissues and cell lines was detected using quantitative real-time PCR. Proliferation and colony forming capacity of BCa cells were assessed using Cell Counting Kit-8, ethynyl-labeled deoxyuridine, and colony formation assays. The cell cycle was analyzed using flow cytometry. Protein expression of insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3) and cyclin dependent kinase 2 (CDK2) was examined using western blots. The binding of RNA and protein was validated using RNA immunoprecipitation. Additionally, xenograft tumor models were established to validate the function of hsa_circ_0005320 in vivo.ResultsWe screened hsa_circ_0005320 from previous high-throughput sequencing and found that it was highly expressed in BCa tissues and associated with tumor differentiation and depth of invasion in BCa patients. Through functional experiments, we demonstrated that hsa_circ_0005320 promoted cell proliferation and regulated the cell cycle. Mechanistically, hsa_circ_0005320 interacted with and upregulated the expression of IGF2BP3, which binds to and enhances the stability of CDK2 mRNA. Furthermore, knockdown of hsa_circ_0005320 resulted in a reduction in tumor burden in vivo.ConclusionsCollectively, these findings highlight the pro-oncogenic role of hsa_circ_0005320 in BCa through the IGF2BP3/CDK2 axis, providing valuable insights into the mechanism of circRNAs in tumor progression.

Resistance to platinum is the main challenge in the chemotherapy of ovarian cancer (OV). Therefore, developing a response signature to platinum is essential for the precision therapy of OV. Existing quantitative signatures of platinum are susceptible to batch effects and sequencing platform variations. To address this, we developed a transcriptome-based platinum signature, named PRSM, consisting of 15 genes, based on within-sample prognostic and relative expression ordering of genes, to predict individual responses to platinum in OV. The PRSM model demonstrated superior classification accuracy compared to previous quantitative signatures. Resistant samples classified by PRSM exhibited poorer overall survival, lower SNV neoantigen load, tumor mutational burden, and distinct methylation patterns compared to sensitive samples. Pathway analysis revealed the activation of MYC targets V2 and oxidative phosphorylation in resistant tumors. Single-cell analysis highlighted the roles of NK and epithelial cells in resistance. Among the 15 core genes, five (TFAP2B, KRT81, PAGE1, CRNN, UGT2B17) were linked to poor prognosis, with TFAP2B having the highest contribution to PRSM. Overexpression of TFAP2B in A2780 cells enhanced cisplatin sensitivity, while in A2780cis cells, it inhibited growth. In brief, our findings provide a multi-dimensional view of platinum resistance in ovarian cancer, introducing a robust predictive model and identifying potential therapeutic targets. ### Competing Interest Statement The authors have declared no competing interest.

Chromothripsis, a hallmark of cancer, is characterized by extensive and localized DNA rearrangements involving one or a few chromosomes. However, its genome-wide frequency and characteristics in urothelial carcinoma (UC) remain largely unknown. Here, by analyzing single-regional and multi-regional whole genome sequencing (WGS), we present the chromothripsis blueprint in 488 UC patients. Chromothripsis events exhibit significant intertumoral heterogeneity, being detected in 41% of UC patients, with an increase from 30% in non-muscle invasive disease (Ta/1) to 53% in muscle-invasive disease (T2-4). The presence of chromothripsis correlates with an unstable cancer genome and poor clinical outcomes. Analysis of multi-regional WGS data from 52 patients revealed pronounced intratumoral heterogeneity with chromothripsis events detectable only in specific tumor regions rather than uniformly across all areas. Chromothripsis events evolve under positive selection and contribute to tumor dissemination. This study presents a comprehensive genome-wide chromothripsis landscape in UC, highlighting the significance of chromothripsis in UC development.

Primordial germ cells (PGCs), as the precursors of gametes found in early embryos, provide a new direction for solving the problem of reproductive disorders. In vitro, conversion of adult stem cells (ASCs) into primordial germ cell-like cells (PGCLCs) is feasible. The means of increasing PGCLCs number in vitro has been a focus of recent stem cell research. In this study, we found that luteinizing hormone (LH) could promote porcine PGCLCs (pPGCLCs) proliferation. To investigate the proliferation regulatory network, whole transcriptome sequencing technology was employed. Results showed that the TGF-β signaling pathway played a key role. In addition, we found that TGFβR1 and SMAD4, TGF-β signaling pathway-related genes, were significantly upregulated after LH treatment. Subsequently, we predicted their target microRNAs (miRNAs) and long non-coding RNAs (lncRNAs): ssc-miR-128, ssc-miR-146b, ssc-miR-361-3p, MSTRG.11473, MSTRG.11475, MSTRG.11553, and MSTRG.11554, and constructed the competitive endogenous RNAs (ceRNA) network. Finally, to further verify the ceRNA network, the miRNA-inhibitors were transfected into cells. RT-qPCR results indicated a significant increase in the expression of MSTRG.11473, MSTRG.11475, MSTRG.11553, MSTRG.11554, TGFβR1, and SMAD4 compared to the negative control (NC) group. In conclusion, these results highlight that LH could regulate the pPGCLCs proliferation by modulating the expression of TGF-β signaling pathway-related ncRNAs.

Breast cancers (BCs) are frequently linked to an immunosuppressive microenvironment that facilitates tumor evasion of anti-cancer immunity. The cells that suppress the immune system such as regulatory B cells (Bregs), regulatory T cells (Tregs), tumor-associated macrophages (TAMs), tumor-associated neutrophils (TANs), myeloid-derived suppressor cells (MDSCs), play a crucial role in immune resistance. Also, tumor progression and immune evasion of cancers are facilitated by cytokines and factors released by tumor cells or immunosuppressive cells. Targeting these regulatory cells therapeutically, whether through elimination, inactivation, or reprogramming, has resulted in hopeful anti-tumor reactions. Yet, the substantial diversity and adaptability of these cells, both in terms of appearance and function, as well as their variation over time and depending on where they are in the body, have posed significant challenges for using them as reliable biomarkers and creating focused therapies that could target their creation, growth, and various tumor-promoting roles. The immunotherapy approaches in BC and their effectiveness in treating certain subtypes are still in their initial phases. In this review, we thoroughly outlined the characteristics, roles, and possible treatment options for these immune-suppressing cells in the tumor environment.

Phospho-H2AX or γ-H2AXis a marker of DNA double-stranded breaks and can therefore be used to monitor DNA repair after, for example, irradiation. In addition, positive staining for phospho-H2AX may indicate genomic instability and telomere dysfunction in tumour cells and tissues. Here, we provide a protocol to perform immunostaining for phospho-H2AX on cells, cryosections and formalinfi xed, paraffi n-embedded tissues. Crucial steps in the protocol and troubleshooting suggestions are indicated. We also provide suggestions on how to combine staining against γ-H2AX with stainings against components of the tumour microenvironment, such as hypoxia and blood vessels.

PDF file - 23K, Figure S3. Map3k2 is a direct target of miR-17/20a as predicted by public algorithms.

[This retracts the article DOI: 10.3892/etm.2017.5528.].

Background A glioma is a tumor originating from glial cells in the central nervous system. Although significant progress has been made in diagnosis and treatment, most high-grade glioma patients are prone to recurrence. Therefore, molecular targeted therapy may become a new direction for adjuvant therapy in glioma. In recent years, many studies have revealed that circular RNA (circRNA) may play an important role in the occurrence and development of many tumors including gliomas. Our previous study found that the expression of hsa_circ_0008922 was up-regulated in glioma tissues upon RNA sequencing. The biological mechanism of circ_0008922 is still unreported in gliomas. Therefore, in this study, we preliminarily outlined the expression of hsa_circ_0008922 in glioma and explored its biological functions. Methods The expression of hsa_circ_0008922 in forty glioma tissues and four glioma cell lines (A172, U251, SF763 and U87) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). The correlation between hsa_circ_0008922 expression and clinicopathological features of glioma patients was evaluated by Fisher’s exact test. To understand the potential function of hsa_circ_0008922 in glioma, we constructed small interfering RNA (siRNA) to hsa_circ_0008922 to downregulate its expression in glioma cell lines A172 and U251. With these hsa_circ_0008922 downregulated cells, a series of assays were carried out as follows. Cell proliferation was detected by CCK8 assay, migration and invasion were determined by wound healing assay and transwell assay, respectively. Colony formation ability was evaluated by plate clonogenic assay. Moreover, flow cytometry combined with Western blot was performed to analyze apoptosis status and the expression of apoptotic related proteins (caspase 3 and caspase 9). Finally, the possible biological pathways and potential miRNA targets of hsa_circ_0008922 were predicted by bioinformatics. Results We found that the expression of hsa_circ_0008922 in glioma tissues was 3.4 times higher than that in normal tissues. The expression of has_circ_0008922 was correlated with WHO tumor grade. After down-regulating the expression of hsa_circ_0008922, malignant biological behavior of glioma cells was inhibited, such as cell proliferation, colony formation, migration, and invasion. At the same time, it also induced apoptosis of glioma cells. Predicted analysis by bioinformatics demonstrated that hsa_circ_0008922 may be involved in tumor-related pathways by acting as a molecular sponge for multiple miRNAs (hsa-let-7e-5p, hsa-miR-506-5p, hsa-let-7b-5p, hsa-let-7c-5p and hsa-let-7a-5p). Finally, we integrated our observation to build a circRNA-miRNA-mRNA predictive network.

Background: Currently, the diagnosis of non-organ-confined bladder cancer (NOCBCa) has a very poor prognosis. For patients with NOCBCa, treatments such as radical cystectomy (RC) and systemic chemotherapy have shown survival benefits. However, the relative survival benefits of trimodal therapy (TMT) are unclear. Methods: Patients diagnosed with NOCBCa (cT4bN0M0, cTxN1-3M0, or TxNxM1) were identified in the Surveillance, Epidemiology, and End Results (SEER) database (2004–2015). Patients were grouped based on their definitive treatment for bladder cancer (RC or TMT with maximal transurethral resection, chemotherapy, or radiotherapy). All-cause mortality (ACM) and bladder cancer-specific mortality (BCSM) were assessed by Cox proportional hazard regression and competitive risk models. Results: A total of 2,988 patients met the inclusion criteria and were treated with RC (83.5%) or TMT (16.5%). Patients who underwent TMT had higher 5-year ACM (91.3%) and BCSM (88.8%) results compared to patients who underwent RC (82.6 and 75.0%, respectively) (P < 0.001). Adjusted hazard rate (AHR) analysis showed that TMT was associated with higher ACM (AHR: 1.33, 95% CI: 1.15–1.54, P < 0.001) and higher BCSM (AHR: 1.32, 95% CI: 1.13–1.54, P = 0.001). Subgroup analysis revealed not statistically significant between RC and TMT among patients aged ≥80 years (P > 0.05). Conclusions: Compared with TMT, RC is associated with a significant reduction in ACM and BCSM. However, the risks and survival benefits of RC should be weighed, especially in older patients, and our results further suggest that there may be no difference in the prognosis of RC and TMT in patients ≥80 years of age. These results are preliminary and emphasize the need for randomized controlled trials to compare TMT and RC.

Bladder cancer (BC) is the ninth most frequent malignancy and the thirteenth leading cause of cancer-related death worldwide. CDKN1A-interacting zinc finger protein 1 (CIZ1) is involved in the development of various cancers, while its role in BC remains unclear. In this study, we found that CIZ1 was up-regulated in BC tissues and cells. Knockdown of CIZ1 suppressed the proliferation and growth of T24 and 5637 cells. Apoptosis and caspase 3/7 activity were enhanced after CIZ1 silencing in both cells. At the molecular level, PCNA, Ki67, HIF-1α, survivin, CCND1 and CCNB1 were down-regulated, and caspase-3, p53, p21, caspase-9 were up-regulated by CIZ1 knockdown. This suggested that silencing of CIZ1 suppressed BC cell proliferation through inducing apoptosis and reducing cell cycle progression. Our study suggests that targeting CIZ1 maybe a potential therapeutic strategy for BC.

MicroRNAs (miRs) serve a role in promoting and suppressing tumors in various types of malignant cancer, such as cervical cancer. However, the regulatory mechanism of miR-124-3p in cervical cancer remains unclear. In the present study, miR-124-3p was significantly downregulated in cervical cancer tissues and cell lines compared with matching adjacent non-tumor tissues and the normal cervical epithelial cell line End1/E6E7, respectively. Decreased expression of miR-124-3p was associated with advanced cervical cancer and the results of an in vitro study demonstrated that the ectopic expression of miR-124-3p significantly decreased the proliferation, migration and invasion of cervical cancer Caski cells. Furthermore, insulin-like growth factor 2 mRNA binding protein 1 (IGF2BP1) was identified as a novel target of miR-124-3p. Overexpression of miR-124-3p decreased the expression of IGF2BP1, whereas miR-124-3p knockdown promoted IGF2BP1 expression at the post-transcriptional level in Caski cells. Additionally, overexpression of IGF2BP1 attenuated the suppressive effects of miR-124-3p on the proliferation, migration and invasion of Caski cells. IGF2BP1 was upregulated in cervical cancer tissues and cell lines compared with matching adjacent non-tumor tissues and the End1/E6E7 cell line, respectively. Therefore, the present study suggests that miR-124-3p suppresses the growth and metastasis of cervical cancer by directly targeting IGF2BP. Thus, miR-124-3p may be developed as a novel method of treating cervical cancer.