恶性肿瘤是由不同群体的癌细胞形成的。癌干细胞（CSCs）是在肿瘤中鉴定出的一个异质细胞亚群，具有自我更新和分化的能力。结直肠癌（CRC）是第三常见的恶性肿瘤，并且有越来越多的证据表明CSCs在癌症发展中至关重要。我们的目标是鉴定CRC细胞和CRC CSCs之间的分子差异，以及这些差异对细胞行为（如迁移、EMT、多能性、形态、细胞周期/控制和表观遗传特征）的影响。我们培养了HT-29细胞系（人结肠腺癌）和HT-29 CSCs（HT-29 CD133+/CD44+细胞）72小时。利用免疫组织化学染色法确定E-钙粘蛋白、KLF4、p53、p21、p16、cyclin D2、HDAC9和P300蛋白的表达水平。采用划痕实验技术评估细胞的迁移能力。此外，利用扫描电子显微镜方法观察细胞的形态特征，并利用EDS比较其周边/中心元素比。此外，使用Muse细胞周期试剂盒来确定细胞周期分析。HT-29 CSC组表现出E-钙粘蛋白、p53、p21、p16、cyclin D2、HDAC9和P300高表达水平，而KLF4在HT-29中高表达。两组在细胞周期各阶段的百分比上没有显著差异。鉴定特定的CSC特征将有助于早期癌症检测和开发更有效的精准肿瘤学选择。© 2023 The Authors. Published by American Chemical Society.

Malignant tumors are formed by diverse groups of cancer cells. Cancer stem cells (CSCs) are a subpopulation of heterogeneous cells identified in tumors that have the ability to self-renew and differentiate. Colorectal cancer (CRC), the third most frequent malignant tumor, is progressively being supported by evidence suggesting that CSCs are crucial in cancer development. We aim to identify molecular differences between CRC cells and CRC CSCs, as well as the effects of those differences on cell behavior in terms of migration, EMT, pluripotency, morphology, cell cycle/control, and epigenetic characteristics. The HT-29 cell line (human colorectal adenocarcinoma) and HT-29 CSCs (HT-29 CD133+/CD44+ cells) were cultured for 72 h. The levels of E-cadherin, KLF4, p53, p21, p16, cyclin D2, HDAC9, and P300 protein expression were determined using immunohistochemistry staining. The migration of cells was assessed by employing the scratch assay technique. Additionally, the scanning electron microscopy method was used to examine the morphological features of the cells, and their peripheral/central elemental ratios were compared with the help of EDS. Furthermore, a Muse cell cycle kit was utilized to determine the cell cycle analysis. The HT-29 CSC group exhibited high levels of expression for E-cadherin, p53, p21, p16, cyclin D2, HDAC9, and P300, whereas KLF4 was found to be high in the HT-29. The two groups did not exhibit any statistically significant differences in the percentages of cell cycle phases. The identification of specific CSC characteristics will allow for earlier cancer detection and the development of more effective precision oncology options.© 2023 The Authors. Published by American Chemical Society.