Leukemia inhibitory factor promotes stemness features of colorectal can-cer cells by modulating CD44 expression
10.3969/j.issn.1000-4718.2024.10.006
- VernacularTitle:白血病抑制因子通过调控CD44的表达增强结直肠癌细胞干性特征
- Author:
Fen QIU
1
;
Xueyan XI
;
Boyu DU
Author Information
1. 湖北医药学院基础医学院免疫教研室,湖北 十堰 442000
- Keywords:
leukemia inhibitory factor;
colorectal cancer;
stemness;
CD44;
ELF3
- From:
Chinese Journal of Pathophysiology
2024;40(10):1826-1833
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To explore the molecular mechanism by which leukemia inhibitory factor(LIF)enhances the stemness characteristics of colorectal cancer(CRC)cells by regulating the expression of the molecular marker CD44 in cancer stem cells(CSCs).METHODS:The expression of LIF in CRC tissues was analyzed using the TCGA public data-base and RNAscope in situ hybridization method.Stable knockdown of LIF was constructed in CRC cell lines(HCT116 and Caco2 cells)using a lentiviral infection system.The experiment was divided into 4 groups:CRC cells control group,CRC cells plus LIF group,CRC cells knockdown control group,and CRC cells knockdown LIF group.The impact of exog-enous LIF on CRC cells' capacity for spheroidization was assessed through stem cell spheroidization assays.The ability of CRC cells to proliferate and migrate was evaluated using MTT,RTCA,plate cloning,and migration tests following the ad-dition or removal of LIF.The impact of LIF on the expression of the transcription factor ELF3 and CD44 in CRC tumor cells was determined using RT-qPCR,a fully automated protein expression analysis system,and Western blot.RT-qPCR was used to identify changes in CD44 splicing variant expression following LIF knockdown.RESULTS:The results dem-onstrated that the expression level of LIF in CRC tissue was higher than that in neighboring cancer tissue(P<0.01),and that patients with CRC who had high expression of LIF had a shorter disease-free survival time(P<0.05).While LIF knockdown can decrease CRC cells' capability to proliferate and migrate(P<0.05),exogenous LIF can boost CRC cells' sphericity,proliferation,and migration capacity(P<0.05).Exogenous LIF can raise(P<0.05)CD44 expression in CRC tumor cells,however LIF reduction can lower(P<0.01)CD44 expression and lower the transcription level of CD44 splic-ing variants.Exogenous injection of LIF or LIF knockdown can cause a significant change(P<0.05)in the expression lev-el of the transcription factor ELF3.CONCLUSION:LIF improves the stemness characteristics of colorectal cancer cells by upregulating the transcription factor ELF3,which also promotes the development of malignancy by increasing CD44 ex-pression in CRC cells.
