Ma Shanshan, Chen Pengfei, Guo Suyang, Wang Liangliang, Hu Jialin, Shao Junjun
Department of Gynecology Oncology, The First Affiliated Hospital of Bengbu Medical University, No.287, Changhuai Road, Longzihu District, Bengbu, 233000, Anhui Province, China.
Department of Thoracic Surgery, The First Affiliated Hospital of Bengbu Medical University, Bengbu, 233000, China.
Biochem Genet. 2024 Nov 22. doi: 10.1007/s10528-024-10945-9.
Ovarian cancer (OC) is a challenging cancer frequently detected at advanced stages. Regulatory B cells (Breg cells) can impair antitumor immunity in patients with OC. The imbalanced serum soluble CD40/CD40L pathway is associated with ovarian tumors. This study aimed to explore the mechanisms involving CD40/CD40L signaling through which Breg cells promote the progression of OC. Breg cells were isolated from peripheral blood samples of 20 patients with OC and 20 healthy controls and identified by flow cytometry. Then, the soluble CD40L concentration in peripheral blood serum of OC patients and healthy volunteers was measured by enzyme-linked immunosorbent assay (ELISA), and we found that the serum soluble CD40L level markedly increased and the proportion of Breg cells was positively correlated with CD40L level in peripheral blood of OC patients. Besides, Breg cells were isolated from spleens of female C57BL/6 WT mice and CD40 mice. Reverse transcription-quantitative polymerase chain reaction, cell counting kit-8 assays, colony formation assays, flow cytometry, Western blotting, wound healing assays, and Transwell assays were conducted to assess the in vitro effect of Breg cells and CD40. We found that Breg cells contributed to cell proliferation, migration, and invasion and suppressed cell apoptosis in OC via the CD40/CD40L pathway. Moreover, we established a xenograft tumor model in female nude BALB/c mice. Tumor size and weight were evaluated, and Western blotting and ELISA were conducted, and we found that Breg cells promoted tumor growth via CD40 signaling. In conclusion, this study demonstrates that Breg cells activated by the CD40/CD40L pathway promotes the aggressiveness of OC cells and tumor growth, indicating that targeting the CD40/CD40L pathway might represent a novel therapeutic option for OC treatment.
卵巢癌（OC）是一种具有挑战性的癌症，常在晚期被发现。调节性B细胞（Breg细胞）可损害OC患者的抗肿瘤免疫力。血清可溶性CD40/CD40L通路失衡与卵巢肿瘤相关。本研究旨在探讨Breg细胞通过CD40/CD40L信号传导促进OC进展的机制。从20例OC患者和20例健康对照的外周血样本中分离Breg细胞，并通过流式细胞术进行鉴定。然后，采用酶联免疫吸附测定（ELISA）检测OC患者和健康志愿者外周血血清中可溶性CD40L浓度，我们发现OC患者外周血中血清可溶性CD40L水平显著升高，且Breg细胞比例与CD40L水平呈正相关。此外，从雌性C57BL/6野生型小鼠和CD40小鼠的脾脏中分离Breg细胞。进行逆转录-定量聚合酶链反应、细胞计数试剂盒-8检测、集落形成检测、流式细胞术、蛋白质免疫印迹法、伤口愈合检测和Transwell检测，以评估Breg细胞和CD40的体外作用。我们发现Breg细胞通过CD40/CD40L通路促进OC细胞增殖、迁移和侵袭，并抑制细胞凋亡。此外，我们在雌性裸BALB/c小鼠中建立了异种移植肿瘤模型。评估肿瘤大小和重量，并进行蛋白质免疫印迹法和ELISA检测，我们发现Breg细胞通过CD40信号传导促进肿瘤生长。总之，本研究表明，由CD40/CD40L通路激活的Breg细胞促进OC细胞的侵袭性和肿瘤生长，表明靶向CD40/CD40L通路可能是OC治疗的一种新的治疗选择。
