Xuandan Qingjin Granule Reversing Osimertinib Resistance by Inhibiting Tumor Stem Cells Through Hh-Ptch/Smo-Gli Regulatory Axis
10.3870/j.issn.1672-0741.24.09.008
- VernacularTitle:玄丹清金颗粒通过Hh-Ptch/Smo-Gli调节轴抑制肿瘤干细胞特性逆转奥希替尼耐药
- Author:
Yang LI
1
;
Lei LIU
;
Jiamao LIN
Author Information
1. 山东第一医科大学附属肿瘤医院山东省精准肿瘤学重点实验室,济南 250117
- Publication Type:Journal Article
- Keywords:
cancer stem cell;
osimertinib;
drug resistance;
Xuandan Qingjin granule
- From:
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
2025;54(1):52-59
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the reversal effect of Xuandan Qingjin(XDQJ)granule on human lung adenocarcinoma PC9/OR osimertinib-resistant cells,and its mechanism.Methods The human lung cancer cell line PC9 was cultivated,and PC9/OR osimertinib-resistant cells were established.CCK-8 was employed to detect the proliferation of PC 9 and PC9/OR trea-ted with different concentrations of Osimertinib,and the effect of different gradient concentrations of XDQJ granule-containing serum on the proliferation of PC9/OR.Western blot and qRT-PCR were utilized to identify the resistance of PC9/OR and the characteristics of tumor stem cells.The soft agar cloning assay was adopted to detect the cell viability.Hoechst33342 and PI double staining were used to determine the proportion of tumor stem cells.Transcriptome sequencing was utilized to screen the related regulatory pathways of tumor stem cells.Western blot was applied to verify the combination of purmorphamine(PM)for reverse validation of tumor stem cell-related regulatory pathways.Results The IC50 of PC9/OR was 45.51 μmol/mL,which was significantly higher than that of PC9(0.07 μmol/mL).The expression levels of p-EGFR,p-ERK,and p-RPS6 of PC9 were significantly decreased after oisimertinib treatment.Compared with PC9,the expression levels of PC9/OR and regulatory mole-cules related to stem cell characteristics were upregulated to varying extents.CCK-8 assay indicated that diluted drug-containing serum with concentration of 40％or less could eliminate cytotoxic interference in rat serum.By means of CCK-8 and cloning ex-periments,it was discovered that the combination of XDQJ granules and Osimertinib could reverse the drug resistance of PC 9/OR.The proportion of stem cells in the combination group was significantly reduced by side group sorting.RNA-seq found sig-nificant changes in PTCH2,SMO,GLI1,GLI2,and GLI3,which are related to tumor stem cell regulation.In the combination group,the survival rate of clonogenic cells increased,OCT4,NANOG,SOX2,SHH,SMO,GLI1 and GLI2 were upregulated,and PTCH2 was downregulated after purmorphamine(PM)treatment.Conclusion XDQJ granule has a potent reversal effect on human lung adenocarcinoma PC9/OR osimertinib-resistant cells,and its mechanism is associated with the inhibition of tumor stem cells by the Hh-Ptch/Smo-Gli regulatory axis.
