It has been popular and challenging to undertake researches on the delay of acquired resistance of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKI). As key cells for tumor initiation, cancer stem cells (CSC) play an important role in the process of resistance to EGFR-TKI. Although preliminary studies found that traditional Chinese medicine (TCM) could inhibit CSC properties and delay EGFR-TKI resistance, the specific molecular mechanism remains unclear. By summarizing the empirical syndrome treatment of EGFR-TKI resistance via TCM and combining recent researches on TCM intervention in CSC to delay EGFR-TKI resistance, this review discussed the potential molecular pathways and mechanisms of deceleration in resistance to EGFR-TKI by TCM via inhibiting CSC characteristics, in order to expand the research ideas of TCM in combination with targeted therapy.
Humans ; Neoplastic Stem Cells/metabolism* ; Drug Resistance, Neoplasm/drug effects* ; ErbB Receptors/genetics* ; Protein Kinase Inhibitors/therapeutic use* ; Medicine, Chinese Traditional ; Neoplasms/drug therapy* ; Animals ; Drugs, Chinese Herbal/therapeutic use*

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.

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.

Epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) is a kind of targeted drug for the treatment of non-small cell lung cancer (NSCLC),but almost all patients develop EGFR-TKI resistance.In Chinese medicine,it is believed that Yang deficiency is the leading cause of tumor,which could end up with the internal formation of cancer toxin and promote the formation of tangible excess pathogens.Clinical and experimental studies have shown that the Chinese medicine treatment based on Yang-warming method can inhibit EGFR-TKI resistance and improve the therapeutic effect of EGFR-TKI.The mechanism may be related to the regulation of miRNA gene and estrogen receptor expression,being against tumor heterogeneity and the regulation of tumor microenvironment.Therefore,Yang deficiency is the key pathogenesis of the EGFR-TKI resistance in NSCLC,and the method reinforcing Yang and suppressing Yin is the basic therapeutic way in Chinese medicine to overcome EGFR-TKI resistance.