Glucose metabolism‐targeted therapy and withaferin A are effective for epidermal growth factor receptor tyrosine kinase inhibitor‐induced drug‐tolerant persisters

Abstract

In pathway‐targeted cancer drug therapies, the relatively rapid emergence of drug‐tolerant persisters (DTPs) substantially limits the overall therapeutic benefit. However, little is known about the roles of DTPs in drug resistance. In this study, we investigated the features of epidermal growth factor receptor–tyrosine kinase inhibitor‐induced DTPs and explored a new treatment strategy to overcome the emergence of these DTPs. We used two EGFR‐mutated lung adenocarcinoma cell lines, PC9 and II‐18. They were treated with 2 μM gefitinib for 6, 12, or 24 days or 6 months. We analyzed the mRNA expression of the stem cell‐related markers by quantitative RT‐PCR and the expression of the cellular senescence‐associated proteins. Then we sorted DTPs according to the expression pattern of CD133 and analyzed the features of sorted cells. Finally, we tried to ablate DTPs by glucose metabolism targeting therapies and a stem‐like cell targeting drug, withaferin A. Drug‐tolerant persisters were composed of at least two types of cells, one with the properties of cancer stem‐like cells (CSCs) and the other with the properties of therapy‐induced senescent (TIS) cells. The CD133high cell population had CSC properties and the CD133low cell population had TIS properties. The CD133low cell population containing TIS cells showed a senescence‐associated secretory phenotype that supported the emergence of the CD133high cell population containing CSCs. Glucose metabolism inhibitors effectively eliminated the CD133low cell population. Withaferin A effectively eliminated the CD133high cell population. The combination of phloretin and withaferin A effectively suppressed gefitinib‐resistant tumor growth.

Overexpression of CD 133 and BCL‐2 in non‐small cell lung cancer with neuroendocrine differentiation after transformation in ALK rearrangement‐positive adenocarcinoma

“…4 The expression of stem cell related markers such as oct3/4 and sox2, was high in cells with high expression of CD133, although we have not yet shown that CD133 directly plays a role on histological transformation. 4 Cancer senescence represents a state of cell cycle arrest. Upregulation of Bcl-2 is reportedly required for the initiation of cell senescence in mouse melanoma cells.…”

“…Exposure to EGFR‐TKIs reportedly also lead to the emergence of drug‐tolerant cells that maintain variability, whereas the majority of the other cell populations typically die in EGFR ‐mutant NSCLC‐derived cell lines . We have previously shown that EGFR‐TKI‐induced drug‐tolerant persisters were composed of two types of cells, cancer stem‐like cells and senescent cells . The expression of stem cell related markers such as oct3/4 and sox2, was high in cells with high expression of CD133, although we have not yet shown that CD133 directly plays a role on histological transformation .…”

“…We have previously shown that EGFR‐TKI‐induced drug‐tolerant persisters were composed of two types of cells, cancer stem‐like cells and senescent cells . The expression of stem cell related markers such as oct3/4 and sox2, was high in cells with high expression of CD133, although we have not yet shown that CD133 directly plays a role on histological transformation . Cancer senescence represents a state of cell cycle arrest.…”

“…In addition, we investigated the associated pathological features before and after transformation. In this case we focus on the cancer stemness as the mechanisms of TKI resistance, because in our previous study we demonstrated the drug‐tolerant clones were consisted of cancer stem cells and senescent cells . Although Giroux‐Leprieur et al .…”

“…In this case we focus on the cancer stemness as the mechanisms of TKI resistance, because in our previous study we demonstrated the drug-tolerant clones were consisted of cancer stem cells and senescent cells. 4 Although Giroux-Leprieur et al reported about the association of cancer stemness and drug resistance in terms of Hedgehog signaling, 5 there are few previous reports about the correlation of cancer stemness and histological transformation.…”