Fig. 7

TM4SF1 could enhance the resistance of HCC to Lenvatinib by upregulating the activation of the NOTCH pathway using MYH9. A Determination of drug resistance in HCC cell lines to lenvatinib using CCK-8. B Using a concentration gradient approach, the Lenvatinib-resistant strain LM3-LR of LM3 cells was cultivated. WB analysis was used to identify the expression of TM4SF1 at various phases of the cultured LM3-LR cells. C Comparison of resistance to lenvatinib between LM3-LR and untreated groups using CCK-8. D The results of the qPCR were utilized to compare the expression of MYH9, molecules associated with cancer stem cells, and molecules connected to the NOTCH pathway in the LM3-LR and untreated groups. E Results of WB analysis were used to compare the expression of MYH9, cancer stem–related molecules, and the NOTCH pathway–related molecules in LM3-LR and untreated groups. F The tumor sphere production experiment revealed that the quantity and diameter of tumor spheres formed by LM3-LR cells were significantly higher than those of the group that had not received any treatment. Scale bars, 50 μm. G A comparison of the fraction of tumor spheres with various sizes and the quantity of tumor spheres. H Tumor spheres were used to extract RNA. After TM4SF1 was silenced, the qPCR analysis revealed that the expression of molecules linked to cancer stem cells, the NOTCH pathway, and MYH9 reduced. I Lenvatinib was administered intraperitoneally before the nude mice were split into the LM3-LR and untreated groups. The cultured nude mice's tumor was removed. J Statistical comparison of tumor volume between LM3-LR and untreated groups. K Statistical comparison of tumor weight between LM3-LR and untreated groups. L Statistical comparison of nude mice weight between LM3-LR and untreated groups. M CCK-8 findings revealed that after TM4SF1 was silenced, LM3-LR's resistance to Lenvatinib diminished. N Efficiency of silencing TM4SF1 in HepG2 was verified via the qPCR analysis. O Efficiency of silencing TM4SF1 in HepG2 was verified via the WB analysis. P CCK-8 findings demonstrated that TM4SF1 silencing reduced HepG2's resistance to Lenvatinib. ^*P < 0.05; ^**P < 0.01; ^***P < 0.001