PO.TB04.03 · 肿瘤生物学
从男性乳腺癌患者来源肿瘤(PDT)中分离、鉴定并建立男性乳腺癌细胞
Isolation, characterization and establishment of male breast cancer cells from male breast cancer patient-derived tumor (PDT)
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
乳腺癌是全球最常见的癌症之一,但在男性中更为致命且更为罕见。目前尚无商业化的男性患者来源乳腺癌细胞系,限制了对男性乳腺癌的研究。因此,由于关于男性乳腺癌的信息有限,大多数研究和治疗方法主要基于女性研究对象。因此,开发男性乳腺癌细胞系将为研究男性特异性乳腺癌提供一个优良的生物学和临床前模型。本研究旨在利用男性乳腺癌患者来源的肿瘤建立新型男性乳腺癌细胞系。乳腺肿瘤取自三位在Simmons癌症研究所接受乳房切除术并知情同意的男性乳腺癌患者。将乳腺肿瘤组织切碎,于37℃过夜酶消化,用无菌滤网过滤,次日离心以沉淀细胞。将沉淀重悬于HUMEC Ready Mix培养基中,培养数周直至贴壁上皮细胞开始生长。使用EpCAM抗体(上皮标志物)通过阳性流式分选纯化上皮细胞群,并在达到第20代时视为永生化。在三个肿瘤样本中,已成功培养出源自两个不同肿瘤样本的两种独特的男性乳腺癌细胞系,分别命名为男性乳腺癌细胞-1(MBCC-1)和男性乳腺癌细胞-2(MBCC-2)。在流式分选期间,目前处于第50代的MBCC-1为70%上皮(EpCAM阳性),而目前处于第19代的MBCC-2细胞为20% EpCAM阳性。MBCC-1的平均圆度和直径分别介于0.6-0.8 uM和12-19微米之间,平均群体倍增时间为42小时。此外,MBCC-1能够通过软琼脂集落形成实验进行非锚定依赖性生长,并对Cytokeratin、CD24、CD44、ALDH1、cKIT、ki-67表现为阳性表达,但对Collagen-IV和成纤维细胞活化蛋白(FAP)表现为阴性。目前正在进行研究以进一步鉴定这两种男性乳腺癌细胞系,包括生物标志物、基因表达谱分析、核型分析、细胞增殖率、软琼脂集落形成实验、营养依赖性检测和致瘤性实验。总之,已建立两种新型男性乳腺癌细胞系,用于研究男性乳腺癌的生物学特性。
查看英文原文 English abstract
Breast cancer is one of the most common cancers worldwide, but it is more fatal and rarer among men. There are no male patient derived breast cancer cell lines commercially available, limiting research on breast cancers in the male patients. Consequently, most research and therapeutic approaches are largely based on female research subjects because of limited information on male mammary carcinoma. Thus, development of male breast cancer cell lines would provide an excellent biological and preclinical model to study male specific mammary carcinoma. This study aims to establish novel male breast cancer cell lines using tumors derived from male mammary carcinoma patients. Mammary tumors were obtained from three consented male breast cancer patients undergoing mastectomy at the Simmons Cancer Institute. The breast tumor tissues were minced, enzymatically digested overnight at 370c, strained using sterile strainer, and centrifuged to pellet the cells on the following day. The pellets were suspended in HUMEC Ready Mix media and cultured for several weeks until adherent epithelial cells started to grow. The epithelial cell populations were purified by positive flow sorting using EpCAM antibody (epithelial marker) and considered immortal upon reaching passage 20. Out of three tumor samples, two unique male breast cancer cell lines from two different tumor samples have been successfully cultured and named as male breast cancer cell-1(MBCC-1) and male breast cancer cell-2 (MBCC-2). During the flow sorting, MBCC-1, which is currently in passage-50, was noted to be 70% epithelial (EpCAM positive) while MBCC-2 cells, which are currently at passage-19, were 20% EpCAM positive. The average circularity and diameter of the MBCC-1 ranges between 0.6-0.8uM and 12-19 microns while the average population doubling time is forty-two hours. Further, MBCC-1 is capable of anchorage independent growth by soft agar colony formation assay and shows positive expression for Cytokeratin, CD24, CD44, ALDH1, cKIT, ki-67 but is negative for Collagen-IV and fibroblast activation protein (FAP) protein. Studies are ongoing to further characterize both the male breast cancer cell lines, including biomarkers, gene expression profiling, karyotyping, cell proliferation rate, soft agar colony formation assay, nutrient dependency tests, and tumorigenicity assays. In summary, two novel male breast cancer cell lines have been established for studying the biology of male breast cancers.
利益披露 Disclosure
S. Kandel, None..
K. Robinson, None..
R. Cosyleon, None..
K. A. Rao, None.