Objective To study the clinical significance of DNA content and expression of nm23-H1, C-erb B-2 and p53 oncoproteins in hepatocellular carcinoma. Methods DNA content was measured after DNA feulgen’s dyeing, the expression of nm23-H1, C-erb B-2 and p53 oncoproteins was detected immunohistochemically. Results The incidence of aneuploid DNA content was 50.0% (12/24) in ≤5 cm group, and was 82.1% (23/28) in ＞5cm group; aneuploid DNA content was related with liver metastasis and cancerous thrombosis. The expression of nm23-H1 in HCC with liver metastasis was higher than that without metastasis. The positive rate of p53 in HCC with canerous thrombosis was higher than that without cancerous thrombosis. The positive rates of nm23-H1 and p53 in HCC with aneuploid DNA content were higher than those with diploid DNA content. The positive rate of C-erb B-2 did not have significantly difference between groups. Postoperative survival rates were possibly related with DNA content as well as expression of nm23-H1, and p53 oncoproteins. Conclusion Aneuploid DNA conten, expression of nm23-H1 and p53 oncoproteins are closely related with the invasiveness of HCC, and have remarkably clinical significance.
Objective To investigate the association of the expression of CD15 mRNA with the invasion and prognosis of hepatocellular carcinoma (HCC) and the expression of nm23H1 mRNA. Methods In situ hybridization and immunohistochemistry methods were used to detect the expression of CD15 mRNA and protein nm23H1 mRNA in HCC.Results In 99 cases of HCC, the positive rate of CD15 mRNA,its protein and nm23H1 mRNA were 38.4%, 36.4% and 76.8%, respectively. The expression of CD15 mRNA was consistent with its protein and negatively correlated with the expression of nm23H1 mRNA. The expression of CD15 mRNA and its protein, nm23H1 mRNA were associated with the invasiveness and metastasis of HCC and the prognosis of HCC patients. Conclusion The detection of CD15 expression could be a new pathological biology index to judge the metastasis and prognosis of HCC.
ObjectiveTo investigate the predicting effect of PIK3CA mutations for the efficacy and prognosis of hepatocellular carcinoma (HCC) patients received surgical resection. MethodsPCR and DNA sequencing were used to detect the PIK3CA mutation status of 79 HCC tissues, its impact on the short and long term effects of the patients were analyzed. ResultsIn this group of patients, mutation rate of PIK3CA gene exon 9 was 39.24% (31/79), PIK3CA mutation rate correlated with lymph node status and tumor differentiation (P < 0.05). The therapeutic effect of patients with PIK3CA mutation was significantly poor than that of the non-mutated group (P < 0.05). The three-year cumulative survival of patients with PIK3CA mutation (33.33%) was significantly lower than non-mutated group's (60.00%) by Kaplan-Meier (P < 0.05). ConclusionPIK3CA gene mutation in exon 9 could impact the efficiency of surgical resection in patients with HCC and could predict a poor survival prognosis.
【Abstract】ObjectiveTo study the mechanism of spontaneous rupture of hepatocellular carcinoma (HCC). MethodsArticles have been reviewed to find out the theory of spontaneous rupture of HCC. ResultsResearchful results suggested that the injury of small arteries was usually followed in patients of spontaneous rupture of HCC. In this review, the immune complex, which composed of hepatitis B virus e antigen, complement C1q and immunoglobulins, was found deposited in the elastic membrane of arteries. Likely as a result of immune complex deposition, vascular injury occurs mainly in the small arteries where the deposition of immune complex was present. The small arteries in which immune complex deposited are readily injuried and cause hemorrhage and rupture of HCC during vascular load increase. ConclusionWe would conclude that immune complex deposition in vessel wall led to the small arteries injury may be the factor involved in the pathogenesis of spontaneous ruptured HCC.
ObjectiveTo identify the risk factors of postoperative recurrence and survival for patients with hepatocellular carcinoma within Milan criteria following liver resection. MethodsData of 267 patients with hepatocellular carcinoma within Milan criteria who received liver resection between 2007 and 2013 in our hospital were retrospectively analyzed. ResultsAmong the 267 patients, 123 patients suffered from recurrence and 51 patients died. The mean time to recurrence were (16.9±14.5) months (2.7-75.1 months), whereas the mean time to death were (27.5±16.4) months (6.1-75.4 months). The recurrence-free survival rates in 1-, 3-, and 5-year after operation was 76.8%, 56.3%, and 47.6%, respectively; whereas the overall survival rates in 1-, 3-, and 5-year after operation was 96.6%, 82.5%, and 74.5%, respectively. Multivariate analyses suggested the tumor differentiation, microvascular invasion, and multiple tumors were independent risk factors for postoperative recurrence; whereas the tumor differentiation, positive preoperative HBV-DNA load, and preoperative neutrophil-to-lymphocyte ratio adversely influenced the postoperative survival. ConclusionsFor patients with hepatocellular carcinoma within Milan criteria after liver resection, the tumor differentiation, microvascular invasion, and multiple tumors contribute to postoperative recurrence; whereas the tumor differentiation, positive preoperative HBV-DNA load, and preoperative neutrophil-to-lymphocyte ratio adversely influence the postoperative survival.
Objective To study the relationship of hepatitis B virus （HBV） to spontaneous rupture of hepatocellular carcinoma （HCC-SR） and its mechanism. Method The related literatures about theory of HCC-SR were consulted and reviewed. Results The injury of small arteries was usually followed in patients with HCC-SR， which was related to vascular autoimmune injury caused by the HBV infection. The small arteries in which immune complex deposited were readily injured， as a result HCC-SR happened while vascular load increased. Conclusion The HBV infection resulted in vascular autoimmune injury maybe a important factor in the pathogenesis of HCC-SR.
Objective To dynamically study the formation of multidrug resistance(MDR) of human hepatocellular carcinoma cell SMMC-7721 induced by Adriamycin (ADM) and the role of multidrug resistance-associated protein(MRP) in its mechanisms.Methods Hepatocellular carcinoma cell SMMC-7721 was cultured in RPMI-1640 medium containing ADM with progressively increased concentration or directly cultured in medium containing different concentrations of ADM. Resistant index of drug-resistant variants of SMMC-7721 cell was determined by drawing cell dosage-reaction curves.Levels of MRP mRNA expression were detected by reverse transcription-polymerase chain reaction(RTPCR). Intracellular rubidomycin(DNR) concentration was examined by flow cytometry(FCM).Results With progressive increasing of ADM concentration in medium resistant index and levels of MRP mRNA expression were correspondingly increased but intracellular DNR concentration was markly reduced. When parental cells were directly cultured in medium containing different concentrations of ADM, the higher the ADM concentration, the higher the level of MRP mRNA expression, but intracellular DNR concentration was kept at the similar high level and most cells died. Conclusion ADM may progressively induce SMMC-7721 cell resistant to multiple chemotherapeutic drugs with reduced intracellular DNR accumulation associated with the overexpression of MRP gene.
To elucidate bcl-2 protein expression in hepatic carcinogenetic process and its relationship with apoptotic changes. bcl-2 protein was evaluated immunohistochemically while apoptosis was approached with terminal deoxynucleotidyl transferase (TdT)mediated dUTP nick end labeling (TUNEL) technique in 8 normal livers (NL), 17 liver cirrhosises (LC) and 77 hepatocellular carcinomas (HCC). The results showed that bcl-2 protein was expressed in 3 of 8 NLs(37.5%), 5 of 17 LCs(29.4%) and 7 of 77 HCCs(9.1%) with significant differences between group NL and HCC and between LC and HCC (Ｐ＜0.05). Apoptosis rates of 1.18±0.42%, 4.85±2.78%, 12.89±2.33% in NL, LC, HCC group respectively were demonstrated with significant differences among them (Ｐ＜0.01). Compared the bcl-2 expression with the apoptosis rate in this hepatocarcinogenetic process, reversed trends were presented. Conclusion: bcl-2 expression could be detected in NL, LC and HCC, and its decreasing expression was related to the inhibition attenuation of hepatocellular apoptosis in the process of hepatocarcinogenesis.
Objective To study the prokaryotic expression of the anti-tumor associated glycoprotein-72 (TAG-72) singlechain fragment variable (scFv) antibody and its specific affinity to hepatocellular carcinoma cell lines and tissues. Methods The cDNA of anti-TAG-72 scFv antibody was inserted into pCANTAB5E to obtain phage vector anti-TAG-72 -scFv-pCANTAB5E. Isopropyl-β-D-thiogalactoside (IPTG) was used to induce the expression of anti-TAG-72 scFv antibody. SDS-PAGE and Western blot were used to identify the anti-TAG-72 scFv antibody. Human hepatocellular carcinoma cells were cultured, and TAG-72 was determined with the obtained scFv by immunohistochemistry in the cells and paraffin-embedded hepatocellular carcinoma tissues. Results SDS-PAGE and Western blot showed that the anti-TAG-72-scFv antibody was successfully expressed. Anti-TAG-72 scFv could bind to hepatocellular carcinoma cell lines SMMC7721, HepG2, and HHCC, but not to BEL7402, suggesting that SMMC7721, HepG2, and HHCC cells expressed TAG-72. For the 40 cases of hepatocellular carcinoma tissues, the positive rate of TAG-72 in stage Ⅰ and Ⅱ-Ⅲ was 23.08% (3/13) and 62.96%(17/27), respectively. While no TAG-72 expression was found in the 10 normal cases. TAG-72 expression was significantly different between hepatocellular carcinomas and normal tissues (Plt;0.05). Conclusions The prokaryotic expression of anti-TAG-72 scFv antibody is successfully achieved, and can be used to identify TAG-72 antigen. TAG-72 is highly expressed in hepatocellular carcinoma, but not in normal liver tissue, which may be suggested as cancer marker of hepatocellular carcinoma.
ObjectiveTo evaluate the relationship between the expression of alpha fetoprotein (AFP) and chemoresistance in hepatocellular carcinoma.MethodsHepatocellular carcinoma was screened from liver tumor tissue samples, which was obtained by puncture before transcatheter arterial chemoembolization (TACE). Immuno-histochemical staining was used to detect the expression of AFP in HCC tissues and the effect of AFP expression in HCC on the effect of chemotherapy was analyzed.ResultsA total of 62 patients met the inclusion criteria, of which 36 were in the chemotherapy resistant group and 26 in the chemotherapy sensitive group. There were 42 patients with positive expression of AFP in tumor tissues (including 29 patients with chemoresistance) and 20 patients with negative expression of AFP in tumor tissues (including 7 patients with chemoresistance). There were no significant difference between the two groups in sex, age, tumor differentiation, Child-Pugh classification of liver function, tumor size, tumor site and hepatitis (P>0.05). In elevated serum AFP level, tumor single, and with portal vein tumor thrombus (PVTT), the proportion of patients in the chemosensitivity group were significantly lower than that in the chemosensitivity group (P<0.05). The results of logistic multivariate regression analysis showed that positive expression of AFP ［OR=0.280, 95%CI (0.092, 0.950), P=0.045］ and PVTT ［OR=0.026, 95%CI (0.004, 0.322), P=0.005］ were independent risk factors for chemotherapeutic resistance in hepatocellular carcinoma.ConclusionAFP positive expression in liver tumor tissues and PVTT are useful indicators of resistance to chemotherapy.