Girotti A W, Hussa R O
Adv Exp Med Biol. 1985;193:129-45. doi: 10.1007/978-1-4613-2165-1_16.
The in vitro phototoxicity of HPD on malignant cells relative to normal cells has been examined. Two human malignant cell lines were studied: the BeWo line of choriocarcinoma cells, which secrete the tumor marker human chorionic gonadotropin (hCG) and its alpha-subunit; and the CaSki line of human cervical carcinoma cells, which secrete hCG and its beta-subunit. Trophoblast-derived, hCG-secreting cells from human amniotic fluid were used as normal controls. In all experiments with HPD plus light, a close correlation was found 24 h after light between cell number and RIA-detectable marker concentration in the medium. Phototoxicity was greater when HPD was introduced in serum-free rather than serum-containing medium. No toxicity was observed in light and dark controls. Cells in Leighton tubes were incubated 1 h with HPD (25 micrograms/ml) in serum-free medium, then rinsed and incubated with medium containing 10% serum. At 2 and 3 days after contact with HPD, flasks were exposed to cool white fluorescent light (1 mW/cm2) for 5 min. Viable cell counts taken 1 day after the light dose indicated that HPD is significantly more phototoxic to BeWo than to CaSki cells; and that both malignant cell types are more photosensitive than amniotic fluid cells, presumably because the latter retain HPD less effectively. In another aspect of this work BeWo cells were used as a model system for comparing the phototoxic effects of the fast (F) and slow (S) eluting fractions of HPD obtained by Bio-Gel P-10 chromatography. Cells in light-shielded tubes were sensitized by incubating with porphyrins for 20 h in media containing 10% calf serum. At 2, 3, or 4 days after removal of porphyrin, with daily replacement of serum-containing medium, flasks were irradiated (see above), and then incubated in the dark for 2 to 4 additional days. Daily culture fluids were analyzed for hormone levels (hCG alpha), and cell counts were performed 2 or 3 days after the light dose. HPD-S (25 micrograms/ml) had no effect on either hormone secretion or cell viability in any of the flasks, whether exposed to light or not. HPD-F at low concentrations (0.25 or 2.5 micrograms/ml) had no detectable effect on cell count or hormone secretion in irradiated flasks. At 10 micrograms/ml, HPD-F was innocuous in dark controls, but caused a large decrease in cell count and hormone output in irradiated flasks.(ABSTRACT TRUNCATED AT 400 WORDS)
已对血卟啉衍生物（HPD）相对于正常细胞对恶性细胞的体外光毒性进行了研究。研究了两种人类恶性细胞系：分泌肿瘤标志物人绒毛膜促性腺激素（hCG）及其α亚基的绒毛膜癌细胞BeWo系；以及分泌hCG及其β亚基的人宫颈癌细胞CaSki系。来自人羊水的滋养层来源、分泌hCG的细胞用作正常对照。在所有HPD加光的实验中，光照后24小时发现细胞数量与培养基中放射免疫分析（RIA）可检测的标志物浓度之间存在密切相关性。当在无血清而非含血清培养基中加入HPD时，光毒性更大。在光照和黑暗对照中未观察到毒性。将Leighton管中的细胞在无血清培养基中与HPD（25微克/毫升）孵育1小时，然后冲洗并在含10%血清的培养基中孵育。在与HPD接触后2天和3天，将培养瓶暴露于冷白色荧光灯下（1毫瓦/平方厘米）5分钟。光照剂量后1天进行的活细胞计数表明，HPD对BeWo细胞的光毒性明显大于对CaSki细胞的光毒性；并且两种恶性细胞类型都比羊水细胞对光更敏感，推测是因为后者保留HPD的效率较低。在这项工作的另一个方面，BeWo细胞被用作模型系统，以比较通过Bio-Gel P-10色谱法获得的HPD的快速（F）和慢速（S）洗脱级分的光毒性作用。将遮光管中的细胞在含10%小牛血清的培养基中与卟啉孵育20小时进行致敏。在去除卟啉后2、3或4天，每天更换含血清培养基，将培养瓶进行照射（见上文），然后在黑暗中再孵育2至4天。每天分析培养液中的激素水平（hCGα），并在光照剂量后2或3天进行细胞计数。HPD-S（25微克/毫升）对任何培养瓶中的激素分泌或细胞活力均无影响，无论是否暴露于光下。低浓度（0.25或2.5微克/毫升）的HPD-F对照射培养瓶中的细胞计数或激素分泌无明显影响。在10微克/毫升时，HPD-F在黑暗对照中无害，但在照射培养瓶中导致细胞计数和激素产量大幅下降。（摘要截取自400字）
