Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation.
Release of monocyte migration signals by breast cancer cell lines after ablative and fractionated γ-irradiation.
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DOI：
10.1186/1748-717x-9-85
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发表时间：
2014-03-26
期刊：
影响因子：
--
通讯作者：
Lauber K
中科院分区：
文献类型：
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作者：
Hennel R;Brix N;Seidl K;Ernst A;Scheithauer H;Belka C;Lauber K
放射治疗，在分割以及消融设置管理，是乳腺癌的一个重要的治疗组成部分。除了直接的肿瘤细胞死亡诱导作用外，越来越多的证据表明免疫机制至少部分有助于其治疗成功。本研究旨在表征分次和消融性放疗诱导的细胞死亡的类型和程度，以及其对死亡乳腺癌细胞释放单核细胞迁移刺激因子的影响。采用细胞死亡和衰老测定来表征具有不同受体和p53状态的一组乳腺癌细胞系对以分次（每日剂量2戈伊）或消融设置（单次剂量20戈伊）施加的γ-辐射的响应。在transwell迁移和2D趋化性/趋化作用测定中检查无细胞培养物上清液的单核细胞迁移刺激潜力。通过qRT-PCR分析辐射诱导的转录应答，并通过流式细胞术测量CD 39表面表达。当以20戈伊的单次消融剂量施加γ-辐射时，具有缺陷p53的快速增殖的激素受体阴性乳腺癌细胞系主要经历原发性坏死，而激素受体阳性的p53野生型细胞显示出凋亡、原发性和继发性（凋亡后）坏死的组合。在坏死过程中，垂死的肿瘤细胞释放腺苷三磷酸双磷酸酶敏感的核苷酸，有效地刺激单核细胞迁移和趋化。在激素受体阳性细胞与功能p53，这是阻碍了辐射诱导的外核苷酸酶CD 39的表面表达。我们的研究表明，消融放射治疗有效地诱导快速增殖，激素受体阴性乳腺癌细胞系突变p53，这反过来释放单核细胞迁移和趋化刺激核苷酸坏死。未来的研究必须阐明，这些机制是否可能被用来刺激肿瘤内单核细胞的募集和随后的适应性抗肿瘤免疫反应的启动，以及哪些乳腺癌亚型可能最适合这种方法。
Radiotherapy, administered in fractionated as well as ablative settings, is an essential treatment component for breast cancer. Besides the direct tumor cell death inducing effects, there is growing evidence that immune mechanisms contribute - at least in part - to its therapeutic success. The present study was designed to characterize the type and the extent of cell death induced by fractionated and ablative radiotherapy as well as its impact on the release of monocyte migration stimulating factors by dying breast cancer cells. Cell death and senescence assays were employed to characterize the response of a panel of breast cancer cell lines with different receptor and p53 status towards γ-irradiation applied in a fractionated (daily doses of 2 Gy) or ablative setting (single dose of 20 Gy). Cell-free culture supernatants were examined for their monocyte migration stimulating potential in transwell migration and 2D chemotaxis/chemokinesis assays. Irradiation-induced transcriptional responses were analyzed by qRT-PCR, and CD39 surface expression was measured by flow cytometry. Fast proliferating, hormone receptor negative breast cancer cell lines with defective p53 predominantly underwent primary necrosis in response to γ-irradiation when applied at a single, ablative dose of 20 Gy, whereas hormone receptor positive, p53 wildtype cells revealed a combination of apoptosis, primary, and secondary (post-apoptotic) necrosis. During necrosis the dying tumor cells released apyrase-sensitive nucleotides, which effectively stimulated monocyte migration and chemokinesis. In hormone receptor positive cells with functional p53 this was hampered by irradiation-induced surface expression of the ectonucleotidase CD39. Our study shows that ablative radiotherapy potently induces necrosis in fast proliferating, hormone receptor negative breast cancer cell lines with mutant p53, which in turn release monocyte migration and chemokinesis stimulating nucleotides. Future studies have to elucidate, whether these mechanisms might be utilized in order to stimulate intra-tumoral monocyte recruitment and subsequent priming of adaptive anti-tumor immune responses, and which breast cancer subtypes might be best suited for such approaches.