Establishment Of Serum ST-PSMD Detecting System And Its Application In HCC Diagnosis

Part ⅠEstablishment of Serum ST-PSMD Detecting System and its Application in HCC DiagnosisBackgroudsHepatocellular carcinoma is one of the most common cancers worldwide.Fifty percent new global HCC cases occur in China annually. Because the most patients with early HCC remain symptomless, they are often diagnosed at intermediate stage or advanced stage. The treatment for liver cancer is limited, and surgical resction is the main option. Moreover, due to high recurrence and metastasis rate after resection the overall and disease-free survival of HCC patients is still disappointing.Therefore, it is urgent to identify novel biomarkers for HCC early detection and therpay. AFP is the only marker widely used for HCC at present. However, accurate rate of AFP in HCC is just between 60%-70% according to previous studies. There is also false positive rate of AFP in benign liver diseases, reproductive teratoma, hepatitis and cirrhosis. Thus far,there are no applicable tumor biomarkers for HCC diagnosis. A great challenge lies ahead in identifying novel biomarkers to increase the accuracy rate of HCC diagnosis. Therefore,the discovery of tumor marker in serum has a great significance.The scientists devote themselves to the study of serum marker, but most studies do not satisfy practical applications. Studing serological marker with high sensitivity and specificity is particularly important.ST-PSMD gene is one of the components of the proteasome which encodes the protein human 26S proteasome subunit non-ATP.ST-PSMD is an important protein of the degradation pathway which can selectively bind to ubiquitinated labeled protein to participate in and mediated protein degradation.The overexpression of ST-PSMD is associated with a variety of autoimmune diseases.Previous studies of our group found that ST-PSMD doesn’t express or express lowly in adjacent normal liver tissues,while mRNA expression and protein of ST-PSMD were all higher in clinical liver samples. ST-PSMD can promote liver cancer cells malignant phenotype, and also can be used as a prognostic marker for HCC.The ST-PSMD application in serological diagnosis of liver cancer markers has not yet been confirmed. Clarifying the value of ST-PSMD in the serological diagnosis of liver cancer by studying this subject and providing the guidance for the clinical diagnosis of liver cancer.ObjectiveEvaluate the value of serum ST-PSMD in the serological diagnosis of HCC by detecting ST-PSMD in patients serum with HCC. Providing the potential markers for the diagnosis of HCC.Methods We collected serum from 100 liver cancer and 20 cirrhosis patients. Serum of 50 healthy donors was also obtatined. Different antibodies were used in ELISA to detect ST-PSMD. And serum ST-PSMD was detected by ELISA. Clinicopathological characteristics were anlayzed and ROC curve was used to measure sensitivity and specificity of ST-PSMD and AFP. Further assessment ST-PSMD and AFP applications in the serological diagnosis of liver cancer.ResultsMatched antibody pairs for ELISA were validated in detecting ST-PSMD. Serum ST-PSMD was detected in clinical specimens by ELISA and the results showed that the serum ST-PSMD in liver cancer patients were much higher than that in the healthy donors and cirrhosis patients(p<0.001). The Maximum of Youden index was used to be the optimal critical value (Cutoff) to identify the three kinds of people. The sensitivity and specificity was 71% and 84.2% respectively,when cutoff value for ST-PSMD was 0.288ng/ml.While the sensitivity and specificity was 60% and 84.2% for AFP with a conventional cutoff value (20ng/ml) suggesting that serum ST-PSMD might serve as a new serological diagnostic biomarker for liver cancer. When combining ST-PSMD and AFP in HCC diagnosis, the sensitivity can be further improved to 80%, specificity was 84.2%.67.5% of patients with serum ST-PSMD positive in HCC patients with AFP-negative, the sensitivity and specificity was 60% and 84.2% for AFP-negative HCC patients,which points that ST-PSMD can improve the diagnosis of AFP negative HCC patients.The serum levels of ST-PSMD were significantly higher in HCC patients with cirrhosis than cirrhosis patients (p<0.001). The results suggest that ST-PSMD is expected to become the differential diagnosis of benign and malignant hepatic disease serological markers.ConclusionThe current study successfully establishes the serological detection system for ST-PSMD using ELISA,and first confirms the serum ST-PSMD in cirrhosis, HCC patients and healthy donors. The results showed that serum levels of ST-PSMD was significantly higher in HCC patients than the healthy donors and patients with cirrhosis (P<0.001).Combining ST-PSMD with AFP can further improve the sensitivity and specificity in serological diagnosis of liver cancer. What’s more, the serum ST-PSMD level of HCC patients with cirrhosis is much higher than that without cirrhosis indicating that serum ST-PSMD can serve as a serological biomarker for the diagnosis of liver cancer.Innovations and potential applicationsThis study successfully establishes the serological detection system for ST-PSMD using ELISA, and first confirms the serum ST-PSMD level in cirrhosis, HCC patients and healthy donors.Combining ST-PSMD with AFP can further improve the sensitivity and specificity in serological diagnosis of HCC.Part ⅡClinical research of primary hepatic neuroendocrine tumorBackgroundPrimary hepatic neuroendocrine tumor (PHNET) is a rare liver cancer. Most patients has late diagnosis and treatment because there is no obvious clinical manifestations. So diagnosis of PHNET must not only rely on pathology and immune phenotype, but also through a comprehensive and long-term follow-up inspection. Just like with primary liver cancer, surgical removal of the tumor is the main treatment. Although some primary hepatic neuroendocrine tumor mass is larger and deeper, surgery still can effectively prolong the survival. One case of huge and rapid progress of primary hepatic neuroendocrine tumor disease-free survival after surgery for up to 4 years has been reported. For patients not suitable for surgical method, liver transplant may be considered. There were reports that patients with endocrine tumors had an example of primary liver nerve disease-free survival after liver transplantation for seven years. Currently, there is no uniform academia histological grading standards.Objective Through analysing 17 cases of primary hepatic neuroendocrine tumors (Primary hepatic neuroendocrine tumor, PHNET) clinical and pathological features and retrospective analysis of clinical data in the Eastern Hepatobiliary Surgery Hospital, clinical knowledge, diagnosis and treatment of the disease can be improved.MethodsAccording to WHO classification of tumors of the digestive system (2010) neuroendocrine tumor grading standards,17 cases of pathological diagnosis of primary hepatic neuroendocrine tumors (PHNET) in our hospital can be clearly classified. The PHNET diagnostic value of this system can be evaluated. By retrospectively analysing the clinical data, the knowledge of PHNET can be raised.Result17 cases of PHNET tissue have been classified by new standard. Five cases failed because the specimen problem. Among the 12 other cases of PHNET,5 cases were NET G1,7 cases were NET G2, zero cases were NEC. All of patients had no significant carcinoid syndrome. NET (G1,G2) had no differences in clinical manifestations. Survival rate was 80% after treatment in NET G1. Survival rate was 57.1% after treatment in NET G2.ConclusionPHNET is a rare liver cancer, and patients often have no typical clinical manifestations. Diagnosis mainly relies on pathological phenotype and classification. The degree of malignancy and prognosis of PHNET needs to digest combined with the WHO classification, the tumor size, the presence of sub-stoves, and other factors to determine.Innovations and potential applicationsEvaluating the WHO classification of tumors of the digestive system (2010) in the diagnosis of neuroendocrine tumor grading standards for primary hepatic neuroendocrine tumors (PHNET).