Differentiation of cancer stem cells into erythroblasts in the presence of CoCl2

Abstract

Cancer stem cells (CSCs) are subpopulations in the malignant tumors that show self-renewal and multilineage differentiation into tumor microenvironment components that drive tumor growth and heterogeneity. In previous studies, our group succeeded in producing a CSC model by treating mouse induced pluripotent stem cells. In the current study, we investigated the potential of CSC differentiation into blood cells under chemical hypoxic conditions using CoCl2. CSCs and miPS-LLCcm cells were cultured for 1 to 7 days in the presence of CoCl2, and the expression of VEGFR1/2, Runx1, c-kit, CD31, CD34, and TER-119 was assessed by RT-qPCR, Western blotting and flow cytometry together with Wright-Giemsa staining and immunocytochemistry. CoCl2 induced significant accumulation of HIF-1α changing the morphology of miPS-LLCcm cells while the morphological change was apparently not related to differentiation. The expression of VEGFR2 and CD31 was suppressed while Runx1 expression was upregulated. The population with hematopoietic markers CD34+ and c-kit+ was immunologically detected in the presence of CoCl2. Additionally, high expression of CD34 and, a marker for erythroblasts, TER-119, was observed. Therefore, CSCs were suggested to differentiate into erythroblasts and erythrocytes under hypoxia. This differentiation potential of CSCs could provide new insight into the tumor microenvironment elucidating tumor heterogenicity.

IL-22 Protects against Biliary Ischemia-Reperfusion Injury after Liver Transplantation via Activating STAT3 and Reducing Apoptosis and Oxidative Stress Levels In Vitro and In Vivo

“…In addition, the expression of IL-22R in HIBEpiCs was detected and confirmed that IL-22R is not expressed in immune cells, indicating that IL-22 could have minimal side effects if used as treatment. It is known that CoCl 2 can induce chemical hypoxia and is ideal for establishing the cellular hypoxia model [ 19 ]. Subsequently, CoCl 2 was used to simulate the hypoxic state of cells in vitro, and the cell viability was detected by the CCK-8 method.…”

Increasing glutathione levels by a novel posttranslational mechanism inhibits neuronal hyperexcitability

“…15 μL of samples were run on 4–20% gradient sodium dodecyl sulfate (SDS) gels (Bio-Rad) at 200V for 35–40min. Proteins were transferred onto polyvinylidene difluoride (PVDF) membranes using a Trans-Blot Turbo™ semi-dry transfer system (Bio-Rad), blocked with 5% bovine serum albumin (BSA, Sigma, MO, USA), and probed with the phospho-S6 ribosomal protein [ [60] , [61] , [62] ] (1:1000, 32 kDa), and β-tubulin [ [63] , [64] , [65] ] (1:1000, 55 kDa) rabbit primary antibodies. β-tubulin was used as a loading control to ensure effective protein transfer and equal sample loading across all wells.…”

“…Proteins were transferred onto PVDF membranes using a Trans-Blot Turbo™ semi-dry transfer system (Bio-Rad), blocked with 5% BSA (Sigma, MO, USA), and probed with the total protein rabbit primary antibodies for TSC1 [ [70] , [71] , [72] ] (1:1000; 150–170 kDa), TSC2 [ [73] , [74] , [75] ] (1:1000; 200 kDa), TBC1D7 [ [76] , [77] , [78] ] (1:1000; 30 kDa). β-tubulin [ [63] , [64] , [65] ] (1:1000; 55kDa) was used as a loading control. Total protein levels of TSC1, TSC2, and TBC1D7 were normalized to the loading control.…”

Cancer stem cells: a target for overcoming therapeutic resistance and relapse

“…CSCs are mostly, but not necessarily found in a mitotically dormant or quiescent state. CSCs can potentially differentiate into different lineage cells, such as cancer cells, vascular endothelial cells, pericytes, and erythroblasts [7][8][9][10] . Recent studies have also revealed the phenotypic and functional heterogeneity of CSCs during tumor progression 11 .…”