A marker for hypoxic cells in tumours with potential clinical applicability.
A marker for hypoxic cells in tumours with potential clinical applicability.
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DOI：
10.1038/bjc.1981.79
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发表时间：
1981-04
影响因子：
8.8
通讯作者：
Sharplin, J
中科院分区：
文献类型：
--
作者：
Chapman, J D;Franko, A J;Sharplin, J
CRAY et al.（1953）提出，肿瘤细胞中的氧浓度可能影响电离辐射对某些人类癌症的治愈能力。最近使用低氧细胞放射增敏剂的临床研究（Urtasun等，1976）、高压氧（Henk & Smith，1977）和贫血患者的治疗前输血（Bush等人，1978）证实了缺氧影响某些肿瘤的放射治疗性的假设。虽然肿瘤的氧合状态被认为是临床肿瘤学家的重要信息，但目前没有技术可用于为个体肿瘤提供此类信息。为测量动物肿瘤中缺氧细胞的分数以及放射治疗过程中它们的再氧合而开发的技术不是常规临床使用，因为它们是侵入性和/或破坏性的（Chapman等人，1981年）。可用于检测实体瘤内缺氧的缺氧细胞放射增敏剂的性质（亚当斯，1973）是硝基芳族药物通过代谢诱导与缺氧细胞的大分子共价结合（McCalla等，1970）和辐射诱导的（Chapman等人，1972年）。目前正在哺乳动物细胞培养物、多细胞球体和动物肿瘤中研究这两种过程的结合机制。用悬浮培养的中国仓鼠V79细胞进行的研究表明，缺氧细胞在37 ℃孵育2小时后结合的~（14）C-misonidazole（MISO）量与几千拉德辐射结合的量相等。随后的努力是针对利用这种代谢诱导的过程标记缺氧细胞在多细胞系统。孵育2小时后，与缺氧细胞结合的14 C-MISO水平会导致每个细胞在14天内发生多次14 C崩解，这种活性应该很容易通过自动射线照相检测到。中国仓鼠V79细胞的多细胞球体在370 C下与50，UM 14 C-MISO（sp. act.）孵育3小时。144 μ Ci/mg，由霍夫曼La Roche，Nutley，NJ慷慨提供）。通过沉淀从标记的培养基中除去球状体，用未标记的盐水洗涤，在10%缓冲福尔马林中固定，包埋并在4 μ m处切片。将安装在显微镜载玻片上的切片浸入液体乳剂（Kodak NTB 3）中并暴露不同的时间长度。乳液显影后，用苏木素对切片进行染色。图1显示了来自一些较小球状体（直径约0-15 mm）的切片的放射自显影（ARC，暴露17天），这些球状体预期不包含缺氧细胞。在该切片的乳剂中可检测到非常少的颗粒，表明从样本中洗涤未结合的14 C-MISO的程序是有效的。图2显示了一个切片的ARG（暴露17天）
CGRAY et al.(1953) suggested that the oxygen concentration intumour cells might influence the curability of some human cancers by ionizing radiation. Recent clinical investigations using hyp-oxic cell radiosensitizers (Urtasun et al., 1976), hyperbaric oxygen (Henk & Smith, 1977) and pretherapy transfusion of anaemic patients (Bush et al., 1978) con-firnm the postulate that lhypoxia doesin-fluence the radiocurability of some tumours. Although the oxygenation status of tumours is considered to be important information for clinical oncologists, no technique is currently available to provide such information for individual tumours. The techniques developed for measuring the fraction of hypoxic cells in animal tumours, and their reoxygenationduring a course ofradiotherapy, are not of routine clinical use because they are invasive and/or destructive (Chapman et al., 1981). A property of hypoxic cell radiosensi-tizers (Adams, 1973), which mightbe exploited for the detectionof hypoxia within solid tumours, is that nitroaromatic drugs become covalently bound to the macromolecules of hypoxic cells by metabolism-induced (McCalla et al., 1970) and radiation-induced (Chapman et al., 1972) processes. Mechanisms of binding by both processes are currently being in-vestigated in mammalian cell cultures, multicellular spheroids and animal tumours. Studies with Chinese hamster V79 cells in suspension culture (to be reported in detailelsewhere) indicated that the amount of 14C-misonidazole (MISO) bound to hypoxic cellsafter 2h incubation at 37 C was equal to that bound by several kilorads of radiation. Subsequent efforts were directed towards exploiting this metabolism-induced pro-cess for labelling hypoxic cells in multicellular systems. The level of 14C-MISO bound to hypoxic cells after 2h incubation cain lead to several disintegrations of 14C per cell in 14 days, an activitywhich should be readily detected by auto-radiography.Multicellular spheroids of Chinese ham-ster V79 cells were incubated for 3 h at 370C with 50, UM 14C-MISO (sp. act. 144, uCi/mg, generously supplied by Hoffman La Roche, Nutley, NJ). The spheroids were removed from the labelled medium by sedimentation, washed with unlabelled saline, fixed in 10% buffered formalin, embedded and sectioned at 4, m. Sections mounted on microscope slides were dipped in liquid emulsion (Kodak NTB3) and exposed for various lengths of time. After development of the emulsion thesections were stainedwith haematoxylin. Fig. 1 shows an autoradiograph (ARC, exposed for 17 days) of a section from some smaller spheroids (-% 0-15 mm in diameter) which would not be expected to contain hypoxic cells. Very few grains can be detected in the emulsion over this section, indicating that the procedures for washing unbound 14C-MISO from the specimen were efficient. Fig. 2 shows an ARG (exposed for 17 days) of a section