The impact of LAG3 splice variants on hepatocellular carcinoma cell proliferation and sorafenib resistance
CHEN Shuai
LIANG Yanwen
LIN Jiansuo
SU Junyu
REN Jianwei
LIU Yi
Abstract：Objective To construct a stable cell line expressing splice variants of immune activation gene 3(LAG3)in hepatocellular carcinoma cells and to investigate the effects of splice variants on the proliferation of hepatocellular carcinoma cells and sorafenib resistance.Methods Hep3B cDNA was used as a template to amplify the LAG3 gene by nested PCR.The differential bands separated by agarose gel electrophoresis were sequenced and compared with the LAG3 transcript variants in the NCBI database.It was found that it lacked exon 3(LAG3 Δ3)relative to the full-length human LAG3 mRNA.The LAG3Δ3-pIGvetor recombinant plasmid was constructed and transfected into Hep3B cells by lentiviral system.After puromycin selection,RT-qPCR and Western blotting were used to verify whether the stable cell line was successfully constructed.The cell proliferation and drug resistance were evaluated by MTT method.Results The levels of LAG3Δ3 mRNA and protein in the selected Hep3B-LAG3Δ3 cells were significantly increased compared with those in the control Hep3B-NC cells(P＜0.000 1).The proliferation ability of cells overexpressing LAG3Δ3 was stronger and their resistance to sorafenib was enhanced compared with Hep3B-NC cells(P＜0.05).Conclusion Hep3B-LAG3Δ3 splicing variant stable cell line was successfully constructed.LAG3Δ3 can promote the proliferation of liver cancer cells and reduce the sensitivity of liver cancer cells to sorafenib.
Keywords：Lymphocyte activation gene 3Splice variantStable transfectionDrug resistanceSorafenib
Publication Date：2026-01-30
Online Publishing Date：2026-03-24(First online date of this platform, not the publication date of the document)
Pages：6( 17-21,95 )