Department of Obstetrics and Gynecology, Center for Reproductive Medicine, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, No.63, Duobao Road, Guangzhou, China.
Key Laboratory for Reproductive Medicine of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Stem Cell Res Ther. 2023 May 4;14(1):119. doi: 10.1186/s13287-023-03348-x.
Cancer stem cells (CSCs) play an important role in endometrial cancer progression and it is potential to isolate CSCs from spheroid cells. Further understanding of spheroid cells at protein level would help find novel CSC markers.
Spheroid cells from endometrial cancer cell lines, Ishikawa and HEC1A, exhibited increased colony forming, subsphere forming, chemo-drug resistance, migration, invasion ability and tumorigenicity, verifying their cancer stem-like cell properties. The up-regulated CD90, CD117, CD133 and W5C5 expression also indicated stemness of spheroid cells. TMT-based quantitative proteomic analysis was performed to explore the potential alterations between parent cells and cancer stem-like spheroid cells. HK2-siRNA was transfected to Ishikawa and HEC1A cells to explore the roles and molecular mechanism of HK2 in endometrial cancer.
We identified and quantified a total of 5735 proteins and 167 overlapped differentially expressed proteins of two cell types, 43 proteins were up-regulated and 124 were down-regulated in spheroid cells comparing with parent cells. KEGG pathway revealed a significant role of HIF-1 pathway in spheroid cells. qRT-PCR and western blot results of GPRC5A, PFKFB3 and HK2 of HIF-1 pathway confirmed their elevated expressions in spheroid cells which were consistent with proteomic results. HK2 promoted cancer stemness in endometrial cancer.
These findings indicate that spheroid cells from endometrial cancer cell lines possess cancer stem-like cell properties and enrich CSCs. HIF-1 pathway is activated in endometrial cancer stem-like spheroid cells.
癌症干细胞（CSC）在子宫内膜癌的进展中起着重要作用，从球体细胞中分离 CSC 是可能的。进一步了解球体细胞在蛋白质水平上的情况有助于发现新的 CSC 标志物。
子宫内膜癌细胞系 Ishikawa 和 HEC1A 的球体细胞表现出增加的集落形成、亚球体形成、化疗药物耐药性、迁移、侵袭能力和致瘤性，验证了它们的癌症干细胞样细胞特性。上调的 CD90、CD117、CD133 和 W5C5 表达也表明了球体细胞的干性。采用 TMT 定量蛋白质组学分析方法探索亲本细胞与癌症干细胞样球体细胞之间的潜在差异。用 HK2-siRNA 转染 Ishikawa 和 HEC1A 细胞，以探讨 HK2 在子宫内膜癌中的作用和分子机制。
我们鉴定和定量了总共 5735 种蛋白质和两种细胞类型的 167 个重叠差异表达蛋白质，与亲本细胞相比，球体细胞中 43 种蛋白质上调，124 种蛋白质下调。KEGG 通路分析显示 HIF-1 通路在球体细胞中具有重要作用。HIF-1 通路中 GPRC5A、PFKFB3 和 HK2 的 qRT-PCR 和 Western blot 结果证实了它们在球体细胞中的高表达，与蛋白质组学结果一致。HK2 促进了子宫内膜癌的癌症干性。
这些发现表明，子宫内膜癌细胞系的球体细胞具有癌症干细胞样细胞特性并富集 CSC。HIF-1 通路在子宫内膜癌干细胞样球体细胞中被激活。
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