Abstract

Cancer stem cells (CSCs) are the most malignant subpopulation of tumor cells that possess a tumorigenic potential and resistantance to chemotherapy. These properties make CSCs a promising target for the development of targeted antitumor therapy which is especially in demand in highly aggressive cancers. However, the correct identification of cancer cells with stem properties remains a challenge. A newly developed lentivirus-based reporter SORE6 allows to directly identify CSCs by measuring gene expression of the embryonic stem cell factors SOX2 and OCT4. In the current study the reporter was modified to enable isolation of SOX2⁺/OCT4⁺ cells by immunomagnetic separation and then was used to transduce HCC1806 and MDA-MB-453 triple-negative breast cancer (TNBC) cell lines. To validate the modified reporter, SOX2⁺/OCT4⁺ populations were isolated and analyzed for the content of NANOG, a key transcription factor of pluropotency which expression is regulated by SOX2/OCT4. The percentage of SOX2⁺/OCT4⁺ cells was assessed for each cell line. An increased content of NANOG protein was found in isolated SOX2⁺/OCT4⁺ cell fractions indicating that the modified reporter is suitable for further studying the CSC subset.

中文翻译：

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改进的基于慢病毒的报告人的癌症干细胞的磁分离。

摘要

癌症干细胞（CSC）是具有致癌潜力和对化疗耐药的肿瘤细胞中最恶性的亚群。这些特性使CSCs成为开发靶向抗肿瘤疗法的有希望的靶标，在高度侵袭性癌症中尤其需要。然而，正确鉴定具有干细胞特性的癌细胞仍然是一个挑战。一种新开发的基于慢病毒的报告基因SORE6可以通过测量胚胎干细胞因子SOX2和OCT4的基因表达来直接鉴定CSC。在当前的研究中，对报告程序进行了修改，以实现SOX2 ⁺ / OCT4 ^+的隔离细胞通过免疫磁分离，然后用于转导HCC1806和MDA-MB-453三阴性乳腺癌（TNBC）细胞系。为了验证修饰的报告基因，分离了SOX2 ⁺ / OCT4 ⁺种群并分析了NANOG的含量，NANOG是多能性的关键转录因子，其表达受SOX2 / OCT4调控。对每种细胞系评估SOX2 ⁺ / OCT4 ⁺细胞的百分比。在分离的SOX2 ⁺ / OCT4 ⁺细胞组分中发现NANOG蛋白含量增加，表明修饰的报道分子适合于进一步研究CSC亚群。