Jinhua Wang

Background: Breast cancer (BC) is the leading malignant tumors among females worldwide, which serves as a common chronic disease with several acute postoperative complications, including upper limb edema, hemorrhage, flap necrosis, effusion and so on. A majority of BC patients have lymph node metastasis, suffering from a poor prognosis. The immune system has been reported to participate in regulating BC lymph node metastasis. This study aimed to search for immune-related biomarkers for predicting BC lymph node metastasis.Methods: 1057 BC patients were acquired from The Cancer Genome Atlas (TCGA) database as the training dataset while 327 BC patients were obtained from GSE20685 as the validation dataset. We get 2,175 immune genes from four immune-related gene sets. We divided BC patients into lymph node positive and negative groups to identify immune-related lymph node-associated differentially expressed genes (DEGs) for functional enrichment analysis and protein-protein interaction (PPI) network. In order to predict BC lymph node metastasis, we established an immune-related signature and assessed its predictive accuracy. In addition, we applied qRT-PCR to investigate signature gene expressions between normal breast epithelium cells and breast cancer cells.Results: We identified 336 immune-related lymph node-associated DEGs, which were enriched in leukocyte migration, immunoglobulin complex and receptor ligand activity among GO analysis and cytokine-cytokine receptor interaction among KEGG analysis. With the aim of predicting BC lymph node metastasis, we established a seven-gene immune-related signature, consisting of F2R, IKZF2, NAB1, RFX5, S100B, S1PR2 and VEGFA. The immune-related signature was proven to be an independent predictive factor for BC lymph node metastasis in both TCGA and GSE20685 databases. Compared with normal breast epithelium cells, RFX5, VEGFA were upregulated in breast cancer cells, IKZF2, NAB1, S100B were downregulated in breast cancer cells while F2R, S1PR2 showed no significance.Conclusion: We established a seven-gene immune-related signature for predicting lymph node metastasis in BC, which might provide a novel sight for BC diagnosis and treatment.

Patients with ovarian cancer (OC) are often diagnosed at an advanced stage and have a poor prognosis because of extensive tumour metastasis. Tumour metastasis usually occurs in stages, which means that before the invasion of tumour cells, a pre-metastatic niche (PMN) has been formed to support the subsequent colonisation and growth of tumour cells. Tumour-associated macrophages (TAMs) are highly heterogeneous in terms of origin, phenotype and function. As one of the most abundant components in the tumour microenvironment (TME), TAMs can contribute to PMN formation by promoting organotropism, mediating immunosuppression, metabolic reprogramming, facilitating angiogenesis, and inducing cancer cell stemness. Therefore, TAMs play an important role in OC metastasis.

This study aimed to investigate the impact of IGF2BP3, a well-known m6A modification-related protein, on the metabolic and immune microenvironment of human cervical cancer. Bioinformatics analysis was performed to analyze the expression of IGF2BP3 in various databases, and its findings were validated using human cervical cancer tissue microarrays. We conducted a study to investigate the impact of IGF2BP3 on glutamine metabolism in cervical cancer cells through the application of metabolomics and metabolic flow analysis. Additionally, we explored how cervical cancer cells promote immune escape by secreting glutamine-derived lactate in a 3D culture setting. To identify the specific targets of IGF2BP3 that influence glutamine metabolism in cervical cancer, we employed RIP-seq analysis. IGF2BP3 exhibited high expression levels in multiple cervical cancer datasets, and its expression was significantly associated with the prognosis of cervical cancer patients. In mixed 3D cell cultures of cervical cancer and T cells, IGF2BP3 was found to enhance glutamate and glutamine metabolism in cervical cancer cells by up regulating the expression of GLS and GLUD1 genes. Moreover, it influenced the differentiation of Treg cells by promoting lactate production and secretion in cervical cancer, leading to immune escape. Mechanistic analysis revealed that IGF2BP3 stabilized the mRNA of GLS and GLUD1 genes through m6A modification, thereby facilitating glutamate and glutamine metabolism in cervical cancer cells and regulating lactate production. Additionally, we investigated the correlation between GLS, GLUD1 protein expression, and IGF2BP3 expression in human cervical cancer through multicolor immunofluorescence staining. The relevance of IGF2BP3 in the context of Treg cell-associated immune escape in cervical cancer was also confirmed. IGF2BP3 exhibits high expression in human cervical cancer and plays a crucial role in stabilizing the mRNA of GLS and GLUD1 genes, key metabolic enzymes in glutamate and glutamine metabolism, through m6A modification. This process leads to immune escape in cervical cancer by promoting lactate production and secretion.

Background: Abnormal N6-methyladenosine (m6A) modification caused by m6A regulators is a common characteristic in various tumors. However, little is known about the role of m6A regulator AlkB homolog 5 (ALKBH5) in triple-negative breast cancer (TNBC). In this study, we analyzed the influence of ALKBH5 on the stemness of TNBC and the molecular mechanism using bioinformatics analysis and in vivo animal experiments. Methods: RNA expression data and single-cell RNA sequencing (scRNA-seq) data were downloaded from the TCGA and GEO databases. Following intersection analysis, key genes involved in the TNBC cell stemness were determined, which was followed by functional enrichment analysis, PPI and survival analysis. Exosomes were extracted from bone marrow mesenchymal stem cells (BMSC-Exos) where ALKBH5 inhibition assay was conducted to verify their function in the biological characteristics of TNBC cells. Results: Bioinformatics analysis revealed 45 key genes of ALKBH5 regulating TNBC cell stemness. In addition, UBE2C was predicted as a key downstream gene and p53 was predicted as a downstream signaling of ALKBH5. In vivo data confirmed that ALKBH5 upregulated UBE2C expression by regulating the m6A modification of UBE2C and reduced p53 expression, thus promoting the stemness, growth and metastasis of TNBC cells. BMSC-Exos suppressed the tumor stemness, growth and metastasis of TNBC cells and ALKBH5 shRNA-loaded BMSC-Exos showed a more significant suppressive role. Conclusion: Taken together, our findings indicated that ALKBH5 shRNA-loaded BMSC-Exos reduced TNBC cell stemness, growth and metastasis and define a promising strategy to treat TNBC.

Objective: To study the efficacy of interval debulking surgery (IDS) plus hyperthermic intraperitoneal chemotherapy (HIPEC) compared to IDS alone for the treatment of ovarian cancer after neoadjuvant chemotherapy (NACT).Methods: We conducted a retrospective study of patients with stage IIIC/IV high-grade serous ovarian carcinoma who underwent surgery at our center from January 2018 to December 2019. Patients who underwent IDS after NACT with (N = 20) or without (N = 42) HIPEC were included. HIPEC was administered after surgery and was combined with 1–2 courses of intraperitoneal hyperthermic perfusion with normal saline only. We analyzed clinical information and outcomes for the two groups.Results: The median progression-free survival (PFS) was 14.05 months in the IDS plus HIPEC group and 12.97 months in the IDS group (P = 0.597). The median overall survival (OS) was not reached. After adjustment for age between the two groups, the differences in PFS and OS remained nonsignificant. The change ratio of postoperative CA-125 to preoperative CA-125 was 0.66 in the IDS plus HIPEC group and 0.53 in the IDS group (P = 0.341). The difference in human epididymis protein 4 (HE-4) change ratio between the two groups was nonsignificant (P = 0.225). No significant difference was observed in the occurrence of grade 3 and 4 adverse events between the two groups (P = 0.201).Conclusion: After NACT, IDS plus HIPEC did not show significant PFS and tumor index change ratio benefits over IDS alone in patients with primary ovarian cancer. Further investigations are needed to assess the role of HIPEC in the treatment of ovarian cancer.

As a result of the limited therapeutic options, advanced cervical cancer is difficult to treat, making the prognosis poor. Therefore, new therapeutic modalities or combinations need to be explored. We herein reported a case of stage IVB cervical cancer which was irresponsive to chemotherapy alone. Based on previous studies and after patient’s consent was obtained, we made a therapeutic plan: chemotherapy (albumin-bound paclitaxel and carboplatin) combined with immunotherapy (PD-1 inhibitor pembrolizumab). After 6 cycles of combined treatment, the patient got almost complete resolution with slight advent event. The treatment was further supported by local radiotherapy combined with immunotherapy. During the treatment period, disease was relatively stable, but the patient suffered severe grade 4 myelosuppression. We were therefore left with no other choice than to interrupt both chemotheraphy and radiotherapy. Before long, the tumor grew explosively again. These guided us to conclude that the combination use of albumin-bound paclitaxel (nab-paclitaxel) and carboplatin and pembrolizumab is effective and well tolerated in the treatment of advanced cervical cancer. The combined use of radiotherapy and pembrolizumab may also be effective. However, the combination use of chemotherapy, radiotherapy and immunotherapy in advanced cancer has not been well studied, and there are still many unsolved queries.

To explore the application value of circulating tumor DNA in the diagnosis of benign and malignant ovarian tumors and its clinical significance in ovarian malignant tumors. A total of 70 patients with ovarian tumors who treated surgery in the first people’s hospital of Lianyungang and the affiliated cancer hospital of Nanjing medical university from January 2017 to August 2020 were collected, including 32 cases of benign tumors, 15 cases of cross-sectional tumors, and 23 cases of malignant tumors. Venous blood was collected before the operation, and the expression level of the plasma circulating tumor DNA was detected by Next-Generation Sequencing, and the expression of circulating tumor DNA chromosome copy number variation in benign and malignant ovarian tumors was analyzed. At the same time, the relationship between the preoperative plasma circulating tumor DNA and clinicopathological features such as tumor size, clinical stage, histological grade, pathological type, and related immunohistochemical indexes were analyzed. There was a significant difference in preoperative plasma circulating tumor DNA among patients with benign, cross-sectional, and malignant tumors of the ovary (p<0.05). In terms of the clinical stage, there were 9 cases in the early stage group (I+II), 5 (55.56 %) cases were positive for plasma circulating tumor DNA before the operation. In the advanced stage group (III+IV), there were 14 cases, 12 (85.71 %) cases were positive for plasma circulating tumor DNA before the operation. In the histological grade of malignant ovarian cancer, there were 6 cases of ovarian low-grade cancer, of which 3 (50.0 %) cases were positive for plasma circulating tumor DNA before the operation. There were 17 cases of ovarian high-grade cancer, of which 14 (82.35 %) cases were positive for plasma circulating tumor DNA before the operation. Among the pathological types of tumors, there were 16 cases of plasma carcinoma and 14 (87.5 %) cases were positive for plasma circulating tumor DNA. There were 7 cases of non-plasma carcinoma and 3 (42.85 %) cases were positive for plasma circulating tumor DNA. In terms of tumor size, there were 7 cases with a diameter <10cm, of which 3 (42.85 %) cases were circulating tumor DNA positive. There were 16 cases of tumor diameter ≥10 cm, of which 14 (87.5 %) cases were circulating tumor DNA positive. In terms of pathological immunohistochemistry, among the 17 ovarian cancer patients with positive preoperative plasma circulating tumor DNA, there were 14 (82.35 %) patients with p53 mutation, and 3 (17.65 %) negative patients. There was no significant difference between the detection rate of plasma circulating tumor DNA and the immunohistochemical indexes such as Ki67, P16, estrogen receptor, progesterone receptor, and Wilms tumor protein-1 before the operation. Circulating tumor DNA level has a high predictive value for the disease progression in patients with ovarian cancer, and can be used as a reference index for clinical monitoring of patients’ condition and prognosis.