Departments of Gynaecology and Obstetrics, The First People's Hospital of Wenling, Wenling, China.
Kaohsiung J Med Sci. 2024 Jun;40(6):530-541. doi: 10.1002/kjm2.12837. Epub 2024 Apr 22.
We previously found that the relative abundance of Bifidobacterium was increased after chemotherapy; however, the role of Bifidobacterium longum in chemotherapeutic drug resistance in ovarian cancer (OVC) remains unclear. This study aimed to understand the potential effects and mechanism of B. longum extracellular vesicles (B. longum-EVs) on carboplatin (CBP) resistance in OVC. Eight normal and 11 ovarian tissues were collected and the expression of B. longum genomic DNA and its association with acquired CBP resistance in OVC patients was determined. After isolating EVs by ultracentrifugation from B. longum (ATCC 15707), CBP-resistant A2780 cells were treated with PBS, CBP, B. longum-EVs, or CBP + B. longum-EVs, and subsequently analyzed by CCK-8, Edu staining, Annexin V/PI double staining, wound healing, and Transwell assays to detect cell viability, proliferation, apoptosis, migration, and invasion, respectively. MRP1, ATP7A, ATP7B, and p53 expression as well as p53 phosphorylation were measured by western blot analysis. S15A mutation of p53 was assessed to examine the potential role of p53 Ser15 phosphorylation in CBP-resistant OVC. B. longum levels were elevated and positively associated with CBP resistance in OVC patients. Only high concentrations of B. longum-EVs attenuated A2780 cell proliferation, apoptosis, migration, and invasion. B. longum-EVs exposure significantly enhanced the sensitivity of CBP-resistant A2780 cells to CBP and decreased the expression of drug resistance-related proteins. The effect of B. longum-EVs on reversing CBP resistance was completely inhibited by S15A mutation of p53. B. longum-EVs enhanced the sensitivity of OVC cells to CBP through p53 phosphorylation on Ser15.
我们之前发现双歧杆菌的相对丰度在化疗后增加；然而，长双歧杆菌在卵巢癌（OVC）化疗药物耐药中的作用尚不清楚。本研究旨在了解长双歧杆菌外囊泡（B. longum-EVs）对 OVC 中卡铂（CBP）耐药的潜在影响和机制。收集了 8 例正常卵巢组织和 11 例卵巢组织，检测长双歧杆菌基因组 DNA 的表达及其与 OVC 患者获得性 CBP 耐药的关系。通过超速离心从长双歧杆菌（ATCC 15707）中分离 EVs 后，用 PBS、CBP、B. longum-EVs 或 CBP+B. longum-EVs 处理 CBP 耐药的 A2780 细胞，随后通过 CCK-8、Edu 染色、Annexin V/PI 双染色、划痕愈合和 Transwell 测定分别检测细胞活力、增殖、凋亡、迁移和侵袭。通过 Western blot 分析检测 MRP1、ATP7A、ATP7B 和 p53 表达以及 p53 磷酸化。评估 p53 S15 突变以检查 p53 Ser15 磷酸化在 CBP 耐药 OVC 中的潜在作用。长双歧杆菌水平在 OVC 患者中升高并与 CBP 耐药呈正相关。只有高浓度的 B. longum-EVs 减弱了 A2780 细胞的增殖、凋亡、迁移和侵袭。B. longum-EVs 暴露显著增强了 CBP 耐药 A2780 细胞对 CBP 的敏感性，并降低了耐药相关蛋白的表达。p53 S15 突变完全抑制了 B. longum-EVs 对逆转 CBP 耐药的作用。B. longum-EVs 通过 p53 Ser15 磷酸化增强 OVC 细胞对 CBP 的敏感性。
