Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; chemically induced ; metabolism ; Cell Transformation, Neoplastic ; Liver ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; beta Catenin ; metabolism

Objective： The current study was designed to search into the detection of micrometastasis in peripheral blood in the patients with hepatocellular carcinoma(HCC)and its significance.Method： Peripheral blood samples were obtained from 65 patients with hepatocellular carcinoma, 21 non-HCC malignant tumors, 22 chronic hepatitis B or cirrhosis, and 21 cases of normal healthy volunteers. For identifying hepatocellular carcinoma cells in peripheral blood, the authors detected liver-specific alpha-fetoprotein(AFP) mRNA from total RNA extracted from whole blood by nested reverse transcription-polymerase chain reaction(Nested-RT-PCR). Results： AFP-mRNA was not detected in the normal healthy volunteers and the patient with non-HCC malignant tumors. The presence of AFP-mRNA in the patients with HCC (67.7％ ,44/65) was higher than those with chronic hepatitis B or cirrhosis ( 9.1％ ,2/22,P< 0.01). The detective rate of AFP-mRNA in the blood seemed to be correlated with the clinical stages of HCC, the presence of extrahepatic metastasis, and portal vein thrombosis. The positive AFP-mRNA in peripheral blood after treatment was correlated with the prognosis of the patients with HCC. AFP-mRNA was detected in 7 of 12 (58.3％) HCC patients with low level of serum AFP (< 25 μg/L). Conclusions： The presence of AFP-mRNA in peripheral blood may be an indicator of malignant or benign hepatocytes, which might predicate hematogenous spreading metastasis of tumor cells in the patients with HCC, it might be a valuable marker for predicting metastasis and recurrence of HCC, and it might play a supplementary role in the diagnosis of HCC with negative or low level of serum AFP.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; Female ; Humans ; Liver Neoplasms ; metabolism ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Receptor, ErbB-2 ; biosynthesis ; genetics ; Receptor, ErbB-3 ; biosynthesis ; genetics

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae

Carcinoma, Hepatocellular ; metabolism ; Glypicans ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; metabolism ; RNA, Messenger ; biosynthesis ; genetics

This research established a simple, rapid and sensitive ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) method to investigate the metabolic profiles of cajanonic acid A (CAA) in rats. After intragastric administration of CAA (30 mg·kg^-1) to rats, the biological samples were detected by UPLC-Q-TOF-MS/MS. Relevant data was collected and processed, the accurate mass and MS² spectra of the metabolites were compared with the parent compound. As a result, a total of 23 metabolites were detected, including 15 in urine, 11 in bile, 11 in feces, and 9 in plasma. The major metabolic pathways related to CAA included dehydrogenation, reduction, hydroxylation, methylation and glucuronide conjugation. This experiment was approved by Animal Ethics Committee of Guizhou Medical University (approval number: 1603137).