Eswaran Sangavi, Bhat Samatha, Upadhya Dinesh, Mascarenhas Roshan, Kabekkodu Shama Prasada
Department of Cell and Molecular Biology, Manipal School of Life Sciences, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India.
Manipal Center for Biotherapeutics Research, Manipal Academy of Higher Education, Manipal, Karnataka, 576104, India.
Sci Rep. 2025 Jan 2;15(1):477. doi: 10.1038/s41598-024-84643-2.
Double C-2 Like Domain Beta (DOC2B) located at 17q13.3 prevents metastasis by senescence induction and epithelial to mesenchymal transition inhibition in cervical cancer (CC). The extracellular vesicle (EV) mediated trafficking of DOC2B and its impact on tumor suppressive activity are not investigated in CC. Using a retroviral method, we first ectopically expressed DOC2B in SiHa, which do not normally express DOC2B. DOC2B-SiHa and vector-SiHa EVs were co-incubated separately with recipient cell and subjected to various cellular and biochemical experiments. For the first time, we demonstrated that DOC2B localizes to EVs, and its transfer to EV may require intracellular calcium. Co-culture of SiHa and HeLa cells with DOC2B-SiHa derived EVs induced morphological changes and suppressed their growth and migration, possibly by induction of G0/G1 to S phase arrest and anoikis. DOC2B-SiHa EVs elevated intracellular reactive oxygen species (ROS) and calcium levels and promoted lipid droplet accumulation and lipid peroxidation rate in recipient cells. DOC2B-SiHa EVs reduced active AKT1 and ERK1/2 levels and EMT marker expression and enhanced cellular senescence and cytotoxic effects of cisplatin. Re-expression of DOC2B significantly altered the global metabolite profile of EVs. Finally, we demonstrated that intracellular calcium chelation significantly reduces DOC2B localization to EVs and impacts its tumor-suppressive properties. Altogether, EV-mediated DOC2B transfer may reduce the aggressive behavior of CC cells.
位于17q13.3的双C-2样结构域β（DOC2B）通过诱导衰老和抑制上皮-间质转化来预防宫颈癌（CC）的转移。在宫颈癌中，尚未研究细胞外囊泡（EV）介导的DOC2B运输及其对肿瘤抑制活性的影响。我们首先使用逆转录病毒方法在通常不表达DOC2B的SiHa细胞中异位表达DOC2B。将DOC2B-SiHa和载体-SiHa细胞外囊泡分别与受体细胞共孵育，并进行各种细胞和生化实验。我们首次证明DOC2B定位于细胞外囊泡，并且其向细胞外囊泡的转移可能需要细胞内钙。用源自DOC2B-SiHa的细胞外囊泡共培养SiHa和HeLa细胞会诱导形态变化，并抑制其生长和迁移，这可能是通过诱导G0/G1期到S期的阻滞和失巢凋亡实现的。DOC2B-SiHa细胞外囊泡提高了受体细胞内的活性氧（ROS）和钙水平，并促进了脂滴积累和脂质过氧化率。DOC2B-SiHa细胞外囊泡降低了活性AKT1和ERK1/2水平以及上皮-间质转化标志物的表达，并增强了细胞衰老和顺铂的细胞毒性作用。DOC2B的重新表达显著改变了细胞外囊泡的整体代谢物谱。最后，我们证明细胞内钙螯合显著降低了DOC2B在细胞外囊泡中的定位，并影响其肿瘤抑制特性。总之，细胞外囊泡介导的DOC2B转移可能会降低宫颈癌细胞的侵袭性。
