Panoply™ Human TNFRSF4 Knockdown Stable Cell Line

Costimulatory molecules that activate T cells play a vital role in immunotherapy. Costimulatory molecules are able to recognize antigenic stimuli required by the immune system. According to previous studies, B7-CD28 and tumor necrosis factor (TNF) families together constitute costimulatory molecules. The B7-CD28 family contains 13 molecules, while the TNF family consists of the TNF receptor superfamily (TNFRSF) and the TNF ligand superfamily (TNFSF), which contain a total of 48 molecules. However, the effects of costimulatory molecules on the carcinogenesis of hepatocellular carcinoma (HCC) have not been elucidated. Here, functional analysis showed that costimulatory molecule genes may be associated with immune-related functions and pathways. Knocking down TNFRSF4 expression significantly reduced the proliferation ability of HCC and enhanced its apoptotic ability. Based on the expression of costimulatory molecules in HCC, the researchers constructed a new risk model, which has good value for predicting prognosis, immune microenvironment, and immunotherapy response. TNFRSF4 has been identified as a potential oncogene in HCC and deserves further study.

TCGA data showed that TNFRSF4 was highly expressed in HCC (Figure 1a). The researchers detected TNFRSF4 expression in 1 normal hepatocyte and 4 HCC cell lines, and found that the expression level of TNFRSF4 in HCC cell lines was high by qRT-PCR (Figure 1b). To further study the mechanism of TNFRSF4, they selected Li-7 and HuH7 cells with high TNFRSF4 expression levels for further experiments. The researchers transfected siRNA targeting TNFRSF4 into Li-7 and HuH7 cells, and detected the transfection efficiency by qRT-PCR (Figure 1c). CCK-8 experiments and colon experiments showed that knocking down TNFRSF4 expression significantly reduced the proliferation ability of Li-7 and HuH7 cells (Figures 1d and e). Flow cytometry analysis showed that the apoptosis ability of Li-7 and HuH7 cells transfected with siTNFRSF4 was enhanced (Figure 1f). In addition, upregulation of Bax protein and downregulation of Bcl-2 protein were observed in TNFRSF4-knockdown HuH7 cells (Figure 1g).

Figure 1. TNFRSF4 was an oncogene in HCC. (Zhou Y, et al., 2022)