| Objectives:To find out the potentially effective and non-invasive biomarkers for monitoring the different disease conditions, the early diagonosis of Hepatocellular carcinoma and prolonging the patients’survival life, serum miR-421and miR-500expression levels were tested using RT-PCR technique from different classification of patients suffering from cirrhosis as well as primary liver cancer. In addition, the levels of amino acids spectrum from serum samples above were investigated in order to lay the foundation for the aided diagnosis of liver disease, nutritional support, as well as the clinical application of amino acid imbalance solution.Methods:Serum samples from16cases of different classifications of cirrohisis which consist of4cases of Child-Pugh A,7cases of Child-Pugh B and5cases of Child-Pugh C hospitalized in liver medicine department;8cases of HCC residents from liver surgical department as well as5cases of normal human from the department of Prevention and Health Care were collected. All of the patients had not received any medical or surgical treatment before admission. All HCC patients were first admitted to hospital with liver lesions and recognised exactly by the Hospital Department of Pathology postoperatively including6cases of middle differentiation and2cases of poor differentiation. MiRNeasy Mini Kit from Qiagen was used to extract serum total RNA and real-time quantitative analysis by RT-PCR was applied. ABI7000system, SPSS17.0and GraphPad Prism4.00were used to analyse the values of serum⊿Ct, AFP, ALT and TBIL levels as well as the correlation between⊿Ct value and AFP, ALT and TBIL values respectively. Amino acid levels of these serum samples were also detected by L-8900Amino Acid Analyzer and the results were analysed by SPSS17.0.Results:(1) Compared with the normal group, serum miR-421and miR-500expression level was not statistically significant in the group of patients with Child Pugh-A liver cirrhosis (P>0.05), but significantly up-regulated in the group of Child-Pugh B cirrhosis, Child-Pugh C cirrhosis and HCC patients (P<0.005). Compared the Child-Pugh B cirrhosis with Child-Pugh C cirrhosis group, serum miR-500level was not significantly different (P>0.05) whereas serum miR-421and miR-500expression levels among other groups were statistically significant (P<0.005).(2) Compared with HCC group, the AFP values decreased stastically in the other groups; the ALT levels are significantly decreased in the normal group and the Child-Pugh A cirrhosis group.Compared to Child-C cirrhosis group, TBIL levels were decreased in normal group and HCC group with statistically significant difference.(3) Relative expression level of miR-421and miR-500(⊿Ct) had significant correlation with AFP; statistically significant correlation with ALT respectively; no significant correlation with TBIL(4) Compared with the normal group, the serum levels of cysteine (P=0.000), glutamate (P=0.048),’methionine (P=0.001), tyrosine (P=0.002) increased and leucine (P=0.049) decreased statistically significantly in the Child-Pugh C cirrhosis group, phenylalanine levels in the Child-Pugh C cirrhosis and HCC groups increased (P=0.000, P=0.001) whereas BIR decreased significantly (P<0.01); compared with Child-Pugh A cirrhosis group, serum isoleucine levels of the Child-Pugh C cirrhosis and HCC patients continued to decline(P=0.005, P=0.006); Compared with Child-Pugh C cirrhosis, phenylalanine levels in the HCC group decreased (P=0.032).Conclusions:This study has found that the level of serum miR-421and miR-500from different Child-Pugh classification of liver cirrhosis and HCC patients are significantly different and monitoring their levels can estimate the disease condition from cirrhosis to HCC, thus they are expected to become potential biomarkers for early diagnosis and treatment of HCC. In addition, serum amino acid levels in different Child-Pugh class cirrhosis and HCC patients also lay the foundation of clinical application for the diagnosis of liver disease, nutritional support and treatment. |
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