Department of Oculoplastic Surgery, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, FL, USA.
Transl Vis Sci Technol. 2021 May 3;10(6):34. doi: 10.1167/tvst.10.6.34.
Sebaceous carcinoma (SC) is a malignant eyelid tumor of the ocular adnexa that is primarily treated via surgical excision. Few therapies exist in advanced cases, and medical therapy is limited because of our incomplete understanding of SC biology. Herein, we describe a technique to culture human ocular adnexal SC for use as an in vitro model.
Human ocular adnexal SC tumor cells were isolated from a patient undergoing orbital exenteration surgery and named Bascom Palmer 50 (BP50). They were cultured in Dulbecco's modified Eagle medium/nutrient mixture F-12 supplemented with 10% fetal bovine serum and antibiotics and were maintained at 37°C in humidified 5% CO2. The cells were characterized by immunohistochemistry, exome sequencing, and short tandem repeats analysis. In vitro drug screening against mitomycin-C (MMC) was performed using a cell viability assay.
BP50 grew past 40 passages with a doubling time of 52.3 hours. Immunocytochemical staining revealed expression of SC-associated markers adipophilin, epithelial membrane antigen, p53, and androgen receptor. Whole exome sequencing showed a significant carryover in somatic mutations between the tumor tissue and corresponding cell line, revealing genetic markers consistent with SC. MMC affected cell viability in a dose-dependent manner.
BP50 displays characteristics of ocular adnexal SC and therefore may facilitate improved understanding of SC biology and the high throughput assessment of novel therapeutic compounds and new drug combinatorial approaches targeted for this disease.
Drug screening with MMC against these cells shows in vitro evidence to support its continued clinical use in SC.
皮脂腺癌（SC）是一种主要通过手术切除治疗的眼附属器恶性眼睑肿瘤。在晚期病例中，几乎没有其他治疗方法，而且由于我们对 SC 生物学的了解不完整，医学治疗也受到限制。在此，我们描述了一种培养人眼附属器 SC 的技术，可将其用作体外模型。
从接受眼眶切除术的患者中分离出人眼附属器 SC 肿瘤细胞，并将其命名为 Bascom Palmer 50（BP50）。它们在补充有 10％胎牛血清和抗生素的 Dulbecco 修改的 Eagle 培养基/营养混合物 F-12 中进行培养，并在 37°C 下于 5％CO2 的潮湿环境中维持。通过免疫组织化学，外显子测序和短串联重复序列分析对细胞进行了表征。使用细胞活力测定法对米托蒽醌（MMC）进行了体外药物筛选。
BP50 生长超过 40 代，倍增时间为 52.3 小时。免疫细胞化学染色显示出与 SC 相关的标志物脂肪细胞素，上皮膜抗原，p53 和雄激素受体的表达。全外显子组测序显示肿瘤组织与相应细胞系之间存在大量体细胞突变，揭示了与 SC 一致的遗传标志物。 MMC 以剂量依赖的方式影响细胞活力。
BP50 表现出眼附属器 SC 的特征，因此可能有助于更好地了解 SC 生物学，并高通量评估针对该疾病的新型治疗化合物和新的药物组合方法。
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