Published online Jun 28, 2026. doi: 10.3748/wjg.v32.i24.118822
Revised: March 8, 2026
Accepted: April 13, 2026
Published online: June 28, 2026
Processing time: 121 Days and 20.1 Hours
Hepatocellular carcinoma (HCC) is a top primary liver malignant tumor, and still is hard to control after invasive growth and spread have already occurred. A big part of its biological difference in properties still cannot get explanation on the molecule level. Transmembrane emp24 domain-containing protein 4 (TMED4) it is one cargo-trafficking protein that belongs to the p24/TMED family. Albumins inside this grouping take part in endoplasmic reticulum-Golgi carrying, and can change signal plans which are related to tumor action, hence the function of TMED4 in HCC has not been clearly given definition. Owing to the fact that nuclear factor kappa-B (NF-κB)/snail pathway has close connection with proliferation, movement ability, invasion capability, and epithelial-mesenchymal transition, hence it is a reasonable path that TMED4 may use it to produce effect on HCC cells.
To describe the expression situation of TMED4 in HCC cells, we therefore confirm whether the forced expression of TMED4 can change cell growth, migration and invasion ability, and we also check whether these appearance changes are accompanied by the change of expression quantity of NF-κB p65 and snail.
The abundance of TMED4 transcription product in LO2 cells and SMMC-7721 cells has been measured through the method of quantitative real-time polymerase chain reaction. Through plasmid transfection, we have constructed a TMED4 overexpression SMMC-7721 model, the empty vector is used as the control by us. The increment of cell number was assessed through cell counting kit-8 test at continuous time points. Transwell chambers that have no or have Matrigel covering were utilized to measure migration and invasion, respectively. We have carried out the operation of Western blot for the detection of the protein expressions of TMED4, NF-κB p65 and snail.
Endogenous TMED4 messenger RNA expression level in SMMC-7721 cells is lower than that in LO2 cells (P < 0.05). After the transfection operation, the group with TMED4 overexpression displayed an obvious elevation in TMED4 messenger RNA when compared with the control group (P < 0.05). Optical density numerical values went up through time in two groups, but the increase was made weaker in TMED4-overexpressing cell groups (P < 0.05). When we make comparison with the control group, the overexpression group was found to have smaller numbers of cells that had migrated and invaded (P < 0.05). The experiment which used Western blot method has discovered that in the cells which have overexpression of TMED4, the protein content of TMED4 and NF-κB p65 is higher, while snail expression level is lower (P < 0.05).
The overexpression of TMED4 has the ability to suppress the proliferation, migration and invasion of HCC cells, therefore it may restrain the biological behaviors of HCC through the regulation of NF-κB/snail signaling pathways.
Core Tip: The transmembrane emp24 domain containing protein 4 (TMED4) expression was lowered in hepatocellular carcinoma (HCC) SMMC-7721 cells when compared with normal LO2 cells. The artificial forced overexpression of TMED4 has obviously held down the proliferation, migration and invasion of HCC cells. From the mechanism perspective, TMED4 made nuclear factor kappa-B p65 have higher expression, and at the same time made snail have lower expression, hence this shows that it restrains malignant phenotypes through adjusting the nuclear factor kappa-B/snail signal axis. These result findings point out that TMED4 is one potential adjustor of HCC aggression capability, and also one candidate molecular target for hereafter treatment exploration.