Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu, Republic of Korea; BK21 Plus KNU Biomedical Convergence Program, Department of Biomedical Science, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.
Exp Cell Res. 2020 Oct 15;395(2):112211. doi: 10.1016/j.yexcr.2020.112211. Epub 2020 Aug 2.
In this study, we noninvasively assessed whether M2-like macrophages accelerate the progression of ovarian cancer by performing molecular imaging of ovarian cancer cells expressing enhanced firefly luciferase (Effluc) in living mice. First, murine ovarian cancer ID8 cells expressing Effluc (ID8/Effluc cells) were established by retroviral infection. Subsequently, macrophages were isolated from the peritoneal exudate of mice injected with thioglycollate medium and differentiated into M2-like macrophages by adding interleukin 4. To characterize these M2-like macrophages, F4/80 and cluster of differentiation 206 expression levels were determined. Then, the M2-like macrophages were co-cultured with the ID8/Effluc cells and bioluminescence imaging (BLI) of signals from the ID8/Effluc cells was completed. Additionally, migration and wound healing were assessed to evaluate the effects of conditioned medium (CM) from M2-like macrophages on ID8/Effluc cell motility. In the in vivo study, mice were first given either liposome-phosphate-buffered saline or liposome-clodronate (lipo-clodronate). After 24 h, ID8/Effluc cells were intraperitoneally injected into the mice and BLI was completed at the designed time points. Next, histological analysis was conducted to characterize the infiltrated tumor. Flow cytometric analysis revealed high levels of CD206 expression in the differentiated M2-like macrophages. Meanwhile, ID8/Effluc cells co-cultured with these M2-like macrophages proliferated rapidly in an M2-like macrophage, number-dependent manner. The migration of the ID8/Effluc cells was also increased by the application of CM from M2-like macrophages. In vivo BLI revealed that the growth rate of intraperitoneally injected ovarian cancer cells was inhibited following macrophage depletion by treatment with lipo-clodronate. M2-like macrophages accelerated the progression of ovarian cancer, suggesting they are a new therapeutic target for ovarian cancer and that ovarian cancer could be managed by altering the nature of communication between ovarian cancer and macrophages.
在这项研究中，我们通过对活鼠体内表达增强型萤火虫荧光素酶（Effluc）的卵巢癌细胞进行分子成像，非侵入性地评估了 M2 样巨噬细胞是否会加速卵巢癌的进展。首先，通过逆转录病毒感染建立了表达 Effluc 的鼠源性卵巢癌细胞（ID8/Effluc 细胞）。随后，从注射巯基乙醇酸盐培养基的小鼠腹腔渗出液中分离巨噬细胞，并通过添加白细胞介素 4 将其分化为 M2 样巨噬细胞。为了鉴定这些 M2 样巨噬细胞，测定了 F4/80 和分化簇 206 的表达水平。然后，将 M2 样巨噬细胞与 ID8/Effluc 细胞共培养，并完成来自 ID8/Effluc 细胞的生物发光成像（BLI）信号。此外，评估了迁移和伤口愈合，以评估 M2 样巨噬细胞条件培养基（CM）对 ID8/Effluc 细胞迁移能力的影响。在体内研究中，首先给小鼠施用脂质体-磷酸盐缓冲盐水或脂质体-氯膦酸盐（脂质体-氯膦酸盐）。24 小时后，将 ID8/Effluc 细胞腹腔内注射到小鼠体内，并在设计的时间点完成 BLI。然后，进行组织学分析以鉴定浸润肿瘤。流式细胞术分析显示，分化的 M2 样巨噬细胞中 CD206 表达水平较高。同时，与这些 M2 样巨噬细胞共培养的 ID8/Effluc 细胞以 M2 样巨噬细胞数量依赖性的方式快速增殖。M2 样巨噬细胞 CM 的应用也增加了 ID8/Effluc 细胞的迁移。体内 BLI 显示，用脂质体-氯膦酸盐处理耗尽巨噬细胞后，注射到腹腔内的卵巢癌细胞的生长速度受到抑制。M2 样巨噬细胞加速了卵巢癌的进展，表明它们是卵巢癌的一个新的治疗靶点，通过改变卵巢癌与巨噬细胞之间的通讯性质，可以对卵巢癌进行管理。
