CDCA2 promotes melanoma progression by inhibiting ubiquitin-mediated degradation of Aurora kinase A.

BACKGROUND: Malignant melanoma is one of the most aggressive types of malignant skin cancer. CDCA2 is of great significance in many tumours, but its role in melanoma is unclear. METHODS: CDCA2 expression in melanoma samples and benign melanocytic naevus tissues was detected by GeneChip and bioinformatics analysis as well as immunohistochemistry. The gene expression in melanoma cells was detected by quantitative PCR detecting system and Western blot. Melanoma models with gene knockdown or overexpression were constructed in vitro, and the effects of gene knockdown or overexpression on melanoma cell phenotype and tumour growth were evaluated by celigo cell counting, transwell, wound healing, flow cytometry and subcutaneous nude mouse tumour models. GeneChip primeview, Ingenuity pathway analysis and bioinformatics analysis combined with co-immunoprecipitation, protein stability experiments and ubiquitination analysis were performed to demonstrate the downstream genes and regulatory mechanism of CDCA2. RESULTS: CDCA2 was highly expressed in melanoma tissues, and CDCA2 level was positively correlated with tumour stage and poor prognosis. CDCA2 downregulation significantly reduced cell migration and proliferation by inducing G1/S phase arrest and apoptosis. CDCA2 knockdown suppressed tumour growth and Ki67 expression in vivo. Mechanistically, CDCA2 inhibited ubiquitin-dependent Aurora kinase A (AURKA) protein degradation by acting on SMAD specific E3 ubiquitin protein ligase 1. AURKA downregulation inhibited melanoma cell proliferation and migration and promoted apoptosis. High expression of AURKA implied poor survival in melanoma patients. Moreover, AURKA knockdown constricted CDCA2 overexpression-induced proliferation and migration. CONCLUSION: CDCA2, which was upregulated in melanoma, enhanced AURKA protein stability by inhibiting SMAD specific E3 ubiquitin protein ligase 1-mediated AURKA ubiquitination, thus playing a carcinogenic role in melanoma progression.

Surgical resection margin for T3-T4 primary acral melanoma: a multicenter retrospective cohort study.

Although the National Comprehensive Cancer Network (NCCN) guidelines include clear recommendations for the appropriate resection margins in non-acral cutaneous melanoma, the required margin for acral melanoma is controversial. In this retrospective study, we aimed to investigate whether narrow-margin excision is warranted for thick acral melanoma. Records from 277 melanoma patients with stage T3-T4 disease who underwent radical surgery in three centers in China from September 2010 to October 2018 were reviewed. Clinicopathologic data, including age, gender, excision margin (1-2 cm versus ≥ 2 cm), Clark level, Breslow thickness, ulceration, N stage and adjuvant therapy, were included for survival analysis. The patients were followed up until death or March 31, 2021. Log-rank and Cox regression analyses were used to identify prognostic factors for overall survival (OS), disease-free survival (DFS) and local and in-transit recurrence-free survival (LITRFS). Among all enrolled patients, 207 (74.7%) had acral melanoma, and 70 (25.3%) had non-acral cutaneous melanoma. No significant difference in baseline characteristics was identified between non-acral and acral melanoma, except for age (p = 0.004), gender (p = 0.009) and ulceration (p = 0.048). In non-acral melanoma, a resection margin of 1-2 cm was a poor independent prognostic factor for OS [p = 0.015; hazard ratio (HR) (95% CI), 0.26 (0.009-0.77)] and LITRFS [p = 0.013; HR (95% CI), 0.19 (0.05-0.71)] but not for DFS [p = 0.143; HR (95% CI), 0.51 (0.21-1.25)]. Forty-three (20.8%) patients in the acral melanoma group had a 1-2-cm resection margin. The resection margin was not correlated with patients' OS (p = 0.196 by log-rank analysis, p = 0.865 by multivariate survival analysis), DFS (p = 0.080 by log-rank analysis, p = 0.758 by multivariate survival analysis) or LITRFS (p = 0.354 by log-rank analysis) in acral melanoma. As recommended in the NCCN guidelines, a resection margin ≥ 2 cm is required for non-acral cutaneous melanoma. Meanwhile, a narrow resection margin (1-2 cm) may be safe for patients with acral melanoma.

Combination of pembrolizumab plus temozolomide therapy in unresectable and advanced melanoma: a multicenter retrospective analysis in China.

BACKGROUND: This study aimed to evaluate the effect of anti-PD-1 combined with temozolomide as front-line therapy in patients with unresectable advanced melanoma. METHODS: The records of patients with unresectable advanced melanoma first treated with pembrolizumab plus temozolomide, pembrolizumab alone, or temozolomide-based chemotherapy at three cancer centers from May 2018 to February 2020 were reviewed. Patients were followed up until death or October 30, 2020. Data were retrospectively reviewed and statistically analyzed for the best objective response rate (ORR) and progression-free survival (PFS), as well as toxicities. RESULTS: Sixty-nine individuals were identified, including 28 (40.6%) with acral melanoma, 18 (26.1%) with cutaneous melanoma, 21 (30.4%) with mucosal melanoma, and two (2.9%) with unknown primary melanoma. The ORR of pembrolizumab plus temozolomide (8/20, 40.0%) in advanced melanoma was higher than pembrolizumab (3/24, 12.5%) and chemotherapy (1/25, 4.0%) alone as front-line therapies. The median PFS of pembrolizumab plus temozolomide as front-line therapy for advanced melanoma was 9.8 months [95% confidence interval (CI): 1.7-17.9 months], which was a significant improvement on the chemotherapy PFS of 4.2 months (95% CI: 2.6-5.8 months) [hazard ratio (HR) 0.415, 95% CI: 0.185-0.931, P=0.033]. The median PFS of pembrolizumab was 6.2 months (95% CI: 2.5-9.9), with no significant difference compared with chemotherapy (HR 0.647, 95% CI: 0.334-1.252, P=0.196). CONCLUSIONS: Combining anti-PD-1 with temozolomide has better efficacy than temozolomide-based chemotherapy or anti-PD-1 alone for advanced melanoma treatment without increasing toxicity. Therefore, anti-PD-1 combined with temozolomide may be preferentially used as a front-line regimen for unresectable advanced melanoma.

The toxicity of cell therapy: Mechanism, manifestations, and challenges.

Adoptive cell therapy (ACT), including tumor-infiltrating lymphocytes (TILs), T cell receptor engineered T cell (TCR-T), and chimeric antigen receptor engineered T cell (CAR-T), has shown significant clinical benefits for cancer treatment. However, all of these ACT therapies are associated with toxicities from mild to life threatening in clinic. Common ACT-related toxicities include cytokine release syndrome (CRS) resulting from immune activation, neurological toxicity, on-target/off tumor or off-target toxicities, and toxicities associated with lymphodepletion preconditioning and high does IL-2 administration. This review summarizes clinical manifestations of adverse events associated with ACT treatment and discusses the underlying pathological mechanisms. Moreover, challenges and opportunities of managing ACT-related toxicities have been discussed to give an indication of how to improve the safety of ACT treatment without dampening the therapeutic effect.

miR-5195-3p Suppresses Cell Proliferation and Induces Apoptosis by Directly Targeting NEDD9 in Osteosarcoma.

Objective: Aberrantly expressed microRNAs (miRs) have associated with the development and progression of osteosarcoma (OS). In this study, the authors aimed to investigate the biological function of miR-5195-3p and the underlying mechanisms. Methods: Quantitative real-time polymerase chain reaction analysis was performed to determine the expression of miR-5195-3p in OS tissues and cell lines. Then, two OS cell lines (MG-63 and U2OS) were transfected with miR-5195-3p mimics to obtain stably miR-5195-3p overexpression cell lines. A series of functional assays, including Cell Counting Kit-8 assay, colony formation assay, flow cytometry assay, and Hoechst staining were performed to analyze cell proliferation and apoptosis. Results: The authors first observed downregulation of miR-5195-3p in OS tissues and cell lines. A series of functional assays demonstrated that miR-5195-3p overexpression significantly attenuated OS cell proliferative activity and induced apoptosis. At a molecular level, the neural precursor cell which expressed developmentally downregulated protein 9 (NEDD9), was inversely correlated with the expression level of miR-5195-3p. Furthermore, ectopic expression of NEDD9 counteracted the antiproliferative and apoptotic effects of miR-5195-3p overexpression in OS cells. Conclusions: In summary, the miR-5195-3p/NEDD9 axis may be a promising antitumor agent for OS.

Anatomical reconstruction of the lateral ligaments of the ankle with semitendinosus allograft.

PURPOSE: The purpose of our study is to evaluate the clinical results of anatomical reconstruction of the lateral ligaments with semitendinosus allograft. METHODS: Thirty-six patients with chronic lateral instability underwent anatomical reconstruction of the lateral ligaments of the ankle with semitendinosus allograft. The American Orthopaedic Foot and Ankle Society Ankle-Hindfoot Scale score (AOFAS score) and the Karlsson score were used to evaluate the clinical results before and after surgery. RESULTS: A total of 35 patients (97.2 %) (36 ankles) were followed up for a mean of 37.9 months. The mean AOFAS score improved from 42.3 ± 4.9 points preoperatively to 90.4 ± 6.7 postoperatively. The mean Karlsson score improved from 38.5 ± 3.2 preoperatively to 90.1 ± 7.8 postoperatively. CONCLUSIONS: Anatomical reconstruction of the lateral ligaments with semitendinosus allograft achieves a satisfactory surgical outcome for chronic ankle instability.