The Function Of TNFSF4 In The Development Of Hepatocellular Carcinoma And The Treatment Of Sorafenib

BackgroundIn recent decades, the prevalence of hepatocellular carcinoma gradually elevates, accounts for over 70% of primary liver cancer, is in the 3rd mortality place among all malignancies. We have achieved great progress in exploring the occurrence and the development of hepatocellular carcinoma in the past decade, however, we still have only limited methods in treating advanced HCC. Sorafenib, a small molecular tyrosine-kinase inhibitor (TKI) targeting VEGFR, PDGFR and Raf kinase family, which can arrest tumor growth in a great spectrum, has been used in clinical for advanced HCC patients. Unfortunately, the response rate of sorafenib is only around 30% and most patients get resistance after 6 months. As a consequence, there’s great demand in researching the resistance of sorafenib.ObjectiveWe explored the function of gene TNFSF4 in hepatocellular carcinoma, and the interaction when treating with sorafenib. We analyzed the express of TNFSF4 in HCC patients, comparing the difference of tumor and para-tumor, to discuss the possible application of TNFSF4 gene in clinical practice.Methods1. Transfect lentivirus in vitro to knockdown TNFSF4 gene in HCC cell lines, to analyze the proteomics change and raise relative hypothesis.2. Testify the function of TNFSF4 gene, including proliferation, invasion, and apoptosis both when TNFSF4 gene was knockdown and when given TNFSF4 cytokine.3. Use sorafenib to simulate the clinical environment, exploring the interaction of TNFSF4 and sorafenib treatment.4. Collect the tissues of HCC patients, analyze the gene expression and protein expression of TNFSF4 using PCR and immunohistochemistry technology respectively.Results1. After knocking down TNFSF4 gene, the expression of JAK-STAT3 was down-regulated, and the expression of cleaved caspase3 was elevated. P53 gene was activated as well as MDM2 decreased. The expression N-Cadherinwas up-regulated.2. The invasion and proliferation of TNFSF4 knockdown cell lines was significantly impaired and apoptosis activated (P<0.001), which could all be reversed by given TNFSF4 cytokine (P<0.001).3. When treating with sorafenib, TNFSF4 gene knockdown could assist the function of sorafenib, the interaction between TNFSF4 gene knockdown and sorafenib treatment was significant (P<0.001). The effect could also be reversed by TNFSF4 cytokine.4. In specimen of HCC patients, the expression of TNFSF4 gene in para-tumor tissue was significantly higher than that in HCC tumor tissue. The result from 3 pairs of immunohistochemistry also was consistent with this.ConclusionsFirstly, Knocking down TNFSF4 gene decreased JAK-STAT3 pathway, promoted p53 expression and restrained MDM2 expression. Then, The knockdown of TNFSF4 gene suppressd the invasion and proliferation in vitro, increased apoptosis, and sensitized the function of Sorafenib, which effects could be reversed by given TNFSF4 cytokine. Finally, The expression of TNFSF4 was higher in para-tumor tissue than that in HCC tumor in gene expression level and consistent with protein expression level.