The presence and extent of regional lymph node and distant metastasis are the most fatal prognostic factors in penile squamous cell carcinoma (PSCC). However, the available biomarkers and detailed mechanisms underlying the metastasis of PSCC remain elusive. Here, we explored the expression landscape of HOX genes in twelve paired PSCC tissues, including primary tumors, metastatic lymph nodes and corresponding normal tissues, and highlighted that HOXD11 was indispensable in the progression of PSCC. HOXD11 was upregulated in PSCC cell lines and tumors, especially in metastatic lymph nodes. High HOXD11 expression was associated with aggressive features, such as advanced pN stages, extranodal extension, pelvic lymph node and distant metastasis, and predicted poor survival. Furthermore, tumorigenesis assays demonstrated that knockdown of HOXD11 not only inhibited the capability of cell proliferation, invasion and tumor growth but also reduced the burden of metastatic lymph nodes. Further mechanistic studies indicated that miR-138-5p was a tumor suppressor in PSCC by inhibiting the translation of HOXD11 post-transcriptionally through binding to the 3′ untranslated region. Furthermore, HOXD11 activated the transcription of FN1 to decompose the extracellular matrix and to promote epithelial mesenchymal transition-like phenotype metastasis via FN1/MMP2/MMP9 pathways. Our study revealed that HOXD11 is a promising prognostic biomarker and predicts advanced disease with poor outcomes, which could serve as a potential therapeutic target for PSCC.

Guidelines recommend intravesical instillation of bacillus Calmette-Guérin (BCG) as the first-choice treatment for intermediate- and high-risk non-muscle-invasive bladder cancer (NMIBC). However, there is no therapeutic biomarker for predicting BCG efficacy, especially in high-risk cases with high failure rates. HER2 expression is considered a prognostic factor for bladder cancer. To elucidate the predictive value and significance of HER2 expression in patients with BCG-exposed NMIBC. A total of 454 patients with NMIBC were included. All patients started BCG intravesical instillation (1.2 × 10⁸ CFU, strain D2PB302) 2-6 wk after transurethral resection of bladder tumor and received 19 treatments over a period of 1 yr. HER2 immunohistochemistry (IHC) results available for 314 patients. The outcomes investigated were recurrence-free survival (RFS) and progression-free survival (PFS). Outcome relationships were explored using multivariable Cox regression and log-rank analysis. In the IHC population, 35.7% of patients had HER2 overexpression (IHC score 2/3+). This group had a poor 5-yr RFS rate of 16.5%, in comparison to 68.0% in the group with low HER2 expression (p < 0.001). Patients with high-risk NMIBC and HER2 overexpression had the highest risk of BCG treatment failure, with 5-yr RFS and PFS rates of 19.0% and 58.2%, respectively. Conversely, HER2-negative (IHC score 0) patients with high-risk NMIBC experienced a long-term BCG benefit, with 5-yr RFS and PFS rates of 80.8% and 92.1%, respectively. Limitations include the retrospective study design and the limited details regarding BCG use. HER2 was an independent predictor of poor BCG efficacy in NMIBC. Patients with high-risk NMIBC and HER2 overexpression had the highest risk of disease recurrence and progression after exposure to BCG. Anti-HER2 targeted therapies could be considered for these patients. Measurement of blood levels of the protein HER2 can be used to predict outcomes after BCG (bacillus Calmette-Guérin) bladder therapy for patients with intermediate- or high-risk non-muscle-invasive bladder cancer. Measurement results for HER2 may help in guiding personalized treatment for these patients.

RAB20, a member of the RAS GTPase oncogene family, is overexpressed in several cancers with poor outcomes, promoting tumorigenesis and inducing genomic instability. Here, we performed comprehensive genomic sequencing on eight penile squamous cell carcinoma (PSCC) and normal tissue pairs and found that RAB20 was upregulated in tumors, especially in metastatic lymph nodes. RAB20 overexpression in tumors was further verified by qPCR, Western blotting, and immunohistochemistry of our newly established PSCC cell lines and paired tissues. The clinical significance of RAB20 was validated in 259 PSCC patients, the largest cohort to date, and high RAB20 expression positively correlated with the T, N, M status, extranodal extension, and clinical stage (all p < 0.01). RAB20 was an unfavorable independent prognostic indicator in the survival analysis (p = 0.011, HR = 2.090; 95% Cl: 1.183–4.692), and PSCC patients with high RAB20 expression experienced shorter 5-year cancer-specific survival times (p < 0.001). Furthermore, tumorigenesis assays demonstrated that RAB20 knockdown inhibited cell proliferation, migration, and colony formation in vitro and tumor growth in vivo. RAB20 depletion also induced PSCC cell cycle arrest at G2/M by increasing Chk1 expression and promoting cdc25c phosphorylation to reduce cdc2-cyclinB1 complex formation. Our study revealed an oncogenic role for RAB20 in promoting PSCC cell proliferation at the G2/M phase via the Chk1/cdc25c/cdc2-cyclinB1 pathway. Thus, RAB20 could be a promising prognostic biomarker of advanced PSCC with poor patient survival outcomes and could be a potential therapeutic target.

Background: Radical inguinal lymph node dissection (rILND) is the most available treatment to cure penile cancer (PC) with limited inguinal-confined disease. However, guidelines regarding acceptable boundaries of rILND are controversial, and consensus is lacking. The authors aimed to standardize the surgical boundaries of rILND with definite pathological evidence and explore the distribution pattern of inguinal lymph nodes (ILNs) in PC. Methods: A total of 414 PC patients from two centers who underwent rILND were enrolled. The ILN distribution was divided into seven zones anatomically for pathological examination. Student’s t test and Kaplan–Meier survival analysis were used. Results: ILNs displayed a funnel-shaped distribution with high density in superior regions. ILNs and metastatic nodes are present anywhere within the radical boundaries. Positive ILNs were mainly concentrated in zone I (51.7%) and zone II (41.3%), but there were 8.7% and 12.3% in inferior zones V and VI, respectively, and 7.1% in the deep ILNs. More importantly, a single positive ILN and first-station positive zone was detected in all seven regions. Single positive ILNs were located in zones I through VI in 40.4%, 23.6%, 6.7%, 18.0%, 4.5%, and 1.1%, respectively, and 5.6% presented deep ILN metastasis directly. Conclusions: The authors established a detailed ILN distribution map and displayed lymphatic drainage patterns with definite pathological evidence using a large cohort of PC patients. Single positive ILNs and first-station metastatic zones were observed in any region, even directly with deep ILN metastasis. Only rILND can ensure tumor-free resection without the omission of positive nodes.

Tumor-associated macrophages (TAMs) are pivotal in facilitating the progression of cancer cells. M1 and M2 are two polarization states of TAMs with opposite functions in tumor progression. While the regulatory role of N6-adenosine (m6A) methylation in macrophage polarization has been established, the function of 5-methylcytosine (m5C) remains unclear. The presence of M2 macrophages in bladder cancer and adjacent normal tissues was validated using fluorescence-activated cell sorting (FACS) and immunofluorescence (IF). The expression of reported m5C regulators (writers, readers, erasers) was screened in M2 macrophages to identify the most relevant regulators. Mechanistic insights into how m5C methylation regulates macrophage polarization were gained through RNA immunoprecipitation (RIP) and quantitative PCR. The FACS and IF results revealed that the main active state of the tumor microenvironment (TME) was the M2 subtype in bladder cancer. Next, NOP2/Sun RNA methyltransferase family member 2 (NSUN2) was identified as the most upregulated RNA m5C methylase in M2 via a qPCR assay. According to whole-transcriptome resequencing in si-NSUN2 RAW/THP-1 cells and GO analysis, SOCS3 was determined to be downstream of NSUN2. By methylating particular sites in SOCS3 mRNA, NSUN2 inhibits both the stability and nuclear export of SOCS3 mRNA, which subsequently activates the JAK2/STAT3 signaling pathway and then promotes macrophage polarization to the M2 phenotype while inhibiting M1 polarization. Additionally, this process involves the assistance and balance of the reader YBX1 and the eraser TET2. NSUN2 methylates SOCS3 mRNA to inhibit its stability and nuclear export, which consequently promotes macrophage polarization to M2.

Aims: To explore the safety and preliminary efficacy of Disitamab Vedotin (DV) combined with Toripalimab (T) and concurrent radiotherapy regimen (DVTR) in the comprehensive bladder-preserving treatment for HER2-positive invasive bladder cancer patients. Material: A total of 10 invasive bladder cancer patients who met the indication for radical cystectomy (RC) but refused the surgery were enrolled from January 2023 to Sep 2025, with follow-up extended until March 2026. All patients underwent maximum transurethral resection of bladder tumor (mTURBT)+/-neoadjuvant therapy with DV plus T after postoperative pathology confirmed HER2 positivity. Concurrent radiotherapy was administered combined with synchronous DV plus T treatment. Patients were followed up until March 2026 or death after the end of treatment. Results: The overall treatment completion rate was 90% (9/10), with 1 patient only receiving 46 Gy of radiotherapy due to personal reasons. The incidence of grade ≥3 adverse reactions during treatment was 10% (1/10), which was leukopenia. The common grade 1–2 adverse reactions were radiation cystitis (9/10) and hand-foot numbness (2/10); hemorrhagic cystitis caused by radiation occurred in 2 patients (2/10) at the 20th and 22nd months after treatment, respectively. At the last follow-up, the median follow-up time was 17.2 months (range, 6–31 months). The bladder preservation success rate was 90% (9/10); the patient who did not complete radiotherapy had T3 recurrence at 5 months, underwent salvage RC and recovered normally, however, died of intestinal obstruction complications 13 months later. The event-free survival rate was 80% (8/10); 1 patient had T1 recurrence at 7 months after treatment and continued bladder-preserving treatment for 31 months. Conclusion: As a novel strategy for comprehensive bladder-preserving treatment, the DVTR regimen based on DV combined with T and concurrent radiotherapy has initially shown acceptable safety and preliminaryly reliable efficacy in this study. It provides a potential bladder-preserving option for patients with HER2-positive invasive bladder cancer regardless of PD-L1 status, though this conclusion requires validation in larger cohorts,which is worthy of further investigation with an expanded sample size.