Influence Of Regorafenib In Hepatocellular Carcinoma Cell Line With High Expression Of Aldolaseb

Background& AimsHepatocellular carcinoma(HCC) is one of the most common malignant tumor with a high cancer-related motality. Portal vein tumor thrombus(PVTT), which is a common complication in patients with advanced-stage HCC, is a major dominant of patients' prognosis. And the treatments of HCC patients with PVTT have not been fundamentally improved in recent years. Therefore, improvements of diagnosis and treatments of HCC with PVTT are clinical priorities. Regorafenib, a novel multi-kinase inhibitor, developed by Bayer Corporation, was used in advanced HCC patients, especially for Sorafenib resistant. Phase II clinical trials have verified the safety and efficacy of Regorafenib in patients with HCC. Our earlier study has demonstrated that patients with high expression of ALDOB, the fourth enzyme of glycolysis, have longer overall survival, but reduced overall survival when Sorafenib being use. Is there an interaction between ALDOB high expression and Regorafenib?Therefore, we have observed HCC metabolic changes through metabonomics, and done researches relating ALDOB and Regorafenib treatment, in order to provide guidance of HCC treatment. MethodsBuild stably expressed cell line using Puromycin resistance plasmid. Observe the influence of ALDOB and Regorafenib on HCC cell in vitro through the CCK8 proliferation experiment, Annexin-V apoptosis experiment, Wound Healing and Transwell migration experiment and verify the results by nude mice subcutaneously tumor-burdened model. Eventually, observe metabolic changes in HCC cells using 13 C isotope labeled glucose tracer experiment. ResultsFunction assay in vitro showed that high expression of ALDOB inhibited cell growth, migration and invasion, and reduced the drug sensitivity of Regorafenib. Highly expressed ALDOB promoted cell growth, migration and invasion, when treated with Regorafenib. Nude mice subcutaneously tumor-burdened model also verified the results. ALDOB over-expression also blocked Regorafenib-mediated apoptosis. The 13 C isotope tracer experiments showed declined metabolism in HCC cells with highly expressed ALDOB, and Regorafenib could lead a decline in metabolism of HCC cells. However, compared with the control group, cells with highly expressed ALDOB showed significantly increased TCA cycle metabolic flow, when treated with Rogerafenib. ConclusionHigh ALDOB expression can suppress HCC cell growth, migration and metabolic. High ALDOB expression can antagonize Regorafenib-mediated growth, apoptosis, migration and metabolic in HCC cells. Advanced liver cancer with highly expressed ALDOB may not be suitable for Regorafenib treatment.