Human Metastatic Melanoma Cell Lines Panel for In Vitro and In Vivo Investigations

E. Kosobokova, N. A. Kalinina, K. M. Konoplina, A. A. Malchenkova, Alexandra E. Evdokimova, Marina V. Piniugina, I. I. Khan, Ilya A. Kislyak, A. Basharina, Anna N. Grishanina, A. A. Rudakova, Pavel O. Varaksa, M. Baryshnikova, V. Pokrovsky, T. A. Bogush, V. Kosorukov
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Abstract

The melanoma origin of cell lines obtained from the axillary lymph node (mel Kas, mel Pet, and mel Lap from patients with a verified diagnosis) was confirmed by the detection of the Melan A melanocyte marker expression. A hyperdiploid (2n+) for the mel Kas line; near-diploid (2n), and in some cells near-tertaploid (4n), and even hypo-octaploid (8n) set (172–179 chromosomes) in the mel Pet cell line; and a hypotetraploid (4n−) for the mel Lap line were detected by karyotypic analysis. All three cell lines are tumorigenic; however, mel Pet demonstrates tumor growth in Balb/c nude mice only in the presence of matrigel. All three lines showed a high expression of TUBB3 and PD-L1 markers, while ERa was low (minimum for mel Pet). Significant differences in the expression level were shown for the Cyt molecular marker. In the transplantation of cells to Balb/c nude mice, a stable expression level is observed only for TUBB3. For the rest of the markers, a decrease in their expression level of varying degrees was noted when the cells were growing in solid tumors in vivo. Mutations were detected in oncogenes (BRAF, EZH2, KIT, KRAS, NRAS, ROS1) and tumor suppressor genes (CDKN2A, FAT4, KMT2C, LRP1B, PTEN, PTPRB, TP53). The detailed characterization of the cell lines makes them valuable for various scientific and regulatory experiments, particularly those involving preclinical data on antiproliferative drugs for malignant melanoma or investigations into melanoma cell properties and progression.
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用于体外和体内研究的人类转移性黑色素瘤细胞系筛选小组
通过检测 Melan A 黑色素细胞标记物的表达,确认了从腋窝淋巴结获得的细胞系(来自确诊患者的 mel Kas、mel Pet 和 mel Lap)的黑色素瘤来源。通过核型分析发现,mel Kas 细胞系为超二倍体(2n+);mel Pet 细胞系为近二倍体(2n),部分细胞近三倍体(4n),甚至低八倍体(8n)(172-179 条染色体);mel Lap 细胞系为低四倍体(4n-)。这三种细胞系都具有致瘤性,但 mel Pet 只有在有 matrigel 的情况下才会在 Balb/c 裸鼠体内生长肿瘤。所有三种细胞系都显示出 TUBB3 和 PD-L1 标记的高表达,而 ERa 的表达较低(mel Pet 最低)。细胞分子标记的表达水平存在显著差异。在将细胞移植到 Balb/c 裸鼠体内时,只观察到 TUBB3 有稳定的表达水平。当细胞在体内实体瘤中生长时,其余标记物的表达水平都出现了不同程度的下降。癌基因(BRAF、EZH2、KIT、KRAS、NRAS、ROS1)和抑癌基因(CDKN2A、FAT4、KMT2C、LRP1B、PTEN、PTPRB、TP53)都发生了突变。这些细胞系的详细特征使它们对各种科学和监管实验,尤其是涉及恶性黑色素瘤抗增殖药物临床前数据或黑色素瘤细胞特性和进展研究的实验具有重要价值。
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