Study Of Adolase B In Sorafenib Resistance Of Hepatocellular Carcinoma

Background &Aims Hepatocellular carcinoma(HCC) is the fifth most common cancer and the third leading cause of cancer death worldwide. The incidence of portal vein tumor thrombus(PVTT) is very high in HCC patients, 50%- 60% of HCC patients find out the PVTT in the first diagnosis by imageology examination. It can lead HCC patients to portal hypertension, deterioration of liver function and within the liver metastatic spread and(or) further, etc. Therefore, early diagnosis and control strategy of clinical HCC with PVTT patient is the overall effect of priority.Sorafenib is recognized as the most effective drug in the treatment of HCC in the world, Adevanced patients is recommended to use Sorafenib was explaned in the Barcelona classification(BCLC). With the clinical application found Sorafenib resistance phenomenon in therapy of clinical, Overdays, using the aspect of cancer stem cells and microenvironment to explain the resistance mechanism, but there are still many limitations. Any genetic or state change will eventually be reflected in the metabolites. Thus metabonomics is an emerging discipline what fromed by proteomics and transcriptome, is the final direction of omics. Also gradually become the cancer research focus on.Therefore, this topic from the clinical phenomenon of using sorafenib HCC with PVTT patients(BCLC: C) is more effective, reveals the law of metabolomics on the metabolic reconstruction, looking for the metabolic reconstruction associated key genes, and illustrate the clinical phenomenon, the more provide guidance for clinical medicationMethods1. Based on13 C stable isotope tracer technology to observe the metabolic flux changes in clinical matching HCC(Normal-Tumor-PVTT) tissues, revealed the characteristics of metabolic reprogramming in HCC with PVTT occurrence and development.2. Contrast gene chip(clinical matching HCC(Normal-Tumor-PVTT) to screen metabolism relative genes. Find out gene ALDOB using Western blot protein immunoblot and immunohistochemical experiment verification.3. Build p GFP-ALDOB puromycin resistance plasmid, packaging by lentivirus transfection Huh-7,MHCC-LM3 and CSQT-2 HCC cells, and obtained stability cell lines by filtering resistance.4. To overexpression ALDOB HCC cell lines as the foundation, through 13 C stable isotope tracer experiment technology to observe ALDOB and with Sorafenib effects on metabolic reprogramming5. To overexpression ALDOB HCC lines as the foundation, using CCK8 assays to study ALDOB and Sorafenib the capacity of proliferation; Application Transwell-migration, Transwell-invasion experiment to study ALDOB and Sorafenib the cell capacity of invasion and metastasis6. Constructs the nude mice subcutaneously tumor-burdened model to observe ALDOB and Sorafenib the tumor-burdened ability in vivo; Build the nude mouse tail vein pulmonary metastasis model to observe ALDOB and Sorafenib internal transfer ability.7. Biological information forecast target protein what can be combined with ALDOB.8. Tissue microarray immunohistochemical analysis of ALDOB and G6 PD expression changes in HCC patients population scale. Using the Chi-square and Student ` s Test analyse expression level of ALDOB`s and G6PD`s relations with clinical pathological characteristics associated index.9. By Kaplan-Meier and Log-rank analyse the relationship ALDOB`s and G6PD`s expression level between clinical prognosis, including: Overall survival time(OS) and disease free survival time(DFS).10. Log-rank univariate analyse prognosis associated clinical pathologic characteristics index, and through the COX regression risk ratios model analysis ALDOB and G6 PD whether as independent risk factors of the prognosis of HCC patients.Results1. Combined clinical matching HCC(Normal-Tumor-PVTT) tissues characteristics of metabolic reprogramming with gene chip(clinical matching HCC(Normal-Tumor-PVTT) tissues) screening and clinical matching HCC tissue sample verification, found in the process of HCC occurrence and development(Normal-Tumor-PVTT) ALDOB gradually decrease.2. After overexpression ALDOB and combine with Sorafenib obviously changes in HCC cell metabolism. Tip: with developing of HCC, ALDOB is the key gene in which regulate reprogramming of glucose metabolism.And overexpression ALDOB use Sorafenib found the opposite characteristic of metaboli reprogramming3. Overexpression ALDOB and sh G6 PD can inhibit the proliferation, combine Tip;ALDOB and G6 PD are the key genes in the development of HCC.overexpression ALDOB with Sorafenib decrease the inhibition bility of proliferation.4. ALDOB inhibits Sorafenib to downgrade of the mitochondrial membrane potential in HCC cells5. ALDOB reduces the Sorafenib inhibition of tumor cells to generate ROS6. ALDOB expression level associated with tumor size, tumor capsule, cancer nest vascular tumor thrombus and tumor cell pathological grade clinical pathological characteristics index. The expression level of G6 PD related clinical pathological characteristics, such as tumor cell pathological grade and recurrence. Tip:ALDOB and G6 PD are playing important role in the process of HCC occurrence development.7. ALDOB`s and G6PD`s expression level is closely related to the prognosis of patients with HCC, ALDOB low expression group of patients’ OS, DFS was lower than that in group ALDOB high expression; High G6 PD express group of patients’ OS, DFS was lower than that in group G6 PD low expression. Factor analysis, ALDOB as independent risk factors of the prognosis of patients with HCC.Conclusions ALDOB participation in the development of HCC glucose metabolic network reprogramming; At the same time involved in regulation of HCC cell proliferation;ALDOB can reduce the inhibitory effect of Sorafenib on tumor cell; ALDOB inhibits Sorafenib to downgrade of the mitochondrial membrane potential in cancer cells.ALDOB reduces the Sorafenib inhibition of tumor cells to generate ROS. In clinical samples ALDOB lower expression; G6 PD high expression. ALDOB`s and G6PD`s expression level are closely related to the clinical pathologic characteristics of patients with HCC. ALDOB`s and G6PD`s expression level are closely related to the prognosis of patients with HCC. ALDOB is independent risk factors of the prognosis of HCC patients. Combined overexpression ALDOB with sh G6 PD deepen inhibit effect in proliferation and metastasis. Combine expression level of ALDOB with G6 PD can better predict the prognosis of patients with HCC. The results suggest:ALDOB and G6 PD can be used as the prognosis index and intervention targets in HCC.