ObjectiveTo study the expression of survivin protein in primary hepatocellular carcinoma(PHC) and its relationship to the proliferation of the tumor cells and prognosis of PHC. MethodsThe expression of survivin protein and the proliferation of tumor cells marked by proliferating cell nuclear antigen (PCNA) in 48 cases of PHC were determined by immunohistochemical method. ResultsThe survivin protein was expressed in 31 of 48 cases of PHC (64.6%). The expression of PCNA was significantly higher in hepatocellular carcinoma (HCC) with positive survivin expression than in HCC with negative survivin expression. The patients with positive survivin expression had the worse prognosis than those with negative survivin expression. ConclusionThe expression of survivin may play an important role in the proliferation of PHC cells and closely associate with the prognosis of PHC, and probably become the prognostic factor and an important target of therapy.
Objective To design and construct the eukaryotic expressed vector of suicide genes, which contained 5 copies of hypoxia-responsive element (5HRE), promoter of alpha-fetoprotein gene (AFPp） and nitroreductase from Escherichia coli. Methods The constructing processes were as follows: ①The design of primer: Suicide genes of NTR in the Escherichia coli, which contained 6his-tag gene (6his-tag), were cloned by overlapping PCR. ②The construction of 5HRE: The single strand of synthetized HRE oligonucleotide was annealed, and 5HRE was constructed by multiple recombinant clone. ③The recombination of NTR gene, 5HRE, AFPp and pIRES2-EGFP: pIRES2-EGFP, which had removed the instant early promoter of cytomegalovirus, was recombined with NTR gene, 5HRE, AFPp. In this way, the eukaryotic expressed vector of pIRES2-EGFP-5HRE-AFPp-NTR, which carried NTR gene, 5HRE, AFPp was finally constructed. Results NTR gene, which contained the fusion of 684-base pair and 6his-tag gene, was cloned successfully, and its sequence was coincident with the result published by Genbank. A 221-base pair of 5HRE was also constructed, which was in accordance with the expected sequences. The integrity of the eukaryotic expressed vector was verified by restriction enzyme digestion and DNA sequence analysis, respectively. Conclusion The eukaryotic expressed vector of pIRES2-EGFP-5HRE-AFPp-NTR is successfully constructed, which may be used for its further investigation in vitro.
Objective To systematically review the efficacy and safety of laparoscopic hepatectomy (LH) and open hepatectomy (OH) for patients with hepatocellular carcinoma (HCC). Methods PubMed, EMbase, The Cochrane Library, CBM, WanFang Data, CNKI databases were electronically searched to collect the case-control studies about LH vs. OH for patients with HCC from inception to December, 2015. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies, then meta-analysis was performed by using RevMan 5.3 software. Results A total of 28 studies involving 1 908 patients were included. The results of meta-analysis showed that: the LH group was superior to OH group on complications (OR=0.35, 95%CI 0.26 to 0.48, P<0.000 01), hospital stay (MD=–4.18, 95%CI (–5.08, –3.29),P<0.000 01), and five years overall survival rate (OR=1.65, 95%CI 1.23 to 2.19,P=0.000 7) and disease-free survival rate (OR=1.51, 95%CI 1.12 to 2.03, P=0.006). However, no significant differences were found in one year and three years overall survival rate, disease-free survival rate, and postoperative recurrence rate. Conclusion Current evidence shows that the LH is superior to OH for the treatment of HCC, and may be amenable to surgery because of its safety and longtime efficacy. Due to limited quality and quantity of the included studies, more high quality studies are needed to verify above conclusion.
【Abstract】Objective To investigate the change of vascular endothelial growth factor (VEGF) expression in HepG2 cells under hypoxia. Methods HepG2 cells were cultured under hypoxia（hypoxia group） and normal condition (control group). VEGF expression of HepG2 cells was examined by immunohistochemical staining. The growth of HepG2 cells was examined by MTT colorimetry and cell count. VEGF level in the culture medium was measured by ELISA.Results After 48 h and 72 h of culture, the growth rate of HepG2 cells in hypoxia group was lower than that in control group (P＜0.05). The cell count in hypoxia group (2.51×104/μl and 2.69×104/μl, respectively) was much lower than that in control group(3.01×104/μl and 3.52×104/μl) after 48h and 72h of culture (P＜0.05). In hypoxia group, VEGF level in the culture medium after 24 h and 48 h was higher than that in control group (P＜0.05, P＜0.01). Conclusion Hypoxia may enhance the VEGF expression in HepG2 cells and this could be the reason of high expression of VEGF after transcatheterized hepatic arterial chemoembolization.
Objective To evaluate the effect of methylation determination about the peripheral plasma DNA in diagnose of hepatocellular carcinoma （HCC） and select the highly sensitive and specific methylated cancer suppressor genes. Methods Methylation-specific PCR （MSP） was used to detect the degree of methylation about SLIT2 and DAPK genes in peripheral plasma and associated cancer tissues of 34 patients with HCC confirmed by pathology, then analyzed their relationship to clinicopathologic feature. Results The positive rate of the promoter methylation of SLIT2 and DAPK genes in cancer tissues in 34 cases were 70.6% （24/34） and 79.4% （27/34）, while the relevant promoter methylation rate in plasma were 44.1% （15/34） and 50.0% （17/34） correspondingly. The sensitivity of detection of DNA methylation about SLIT2 and DAPK genes in plasma was 62.5% and 63.0%, respectively；both of the specificity for them were 100%. The negative predicted value was 52.6% and 41.2%, respectively；while both of the positive predicted value were 100%. There were no significant correlation between the clinicopathologic features and the methylation rate in cancer tissues and plasma （P＞0.05）. In plasma of patients whose AFP＜400 μg/L, the positive rate of combined detection of DNA methylation of SLIT2 and DAPK was 61.1% （11/18）. Conclusions The detection rate of DNA methylation of SLIT2 and DAPK genes in plasma is higher, and there is a significant correlation between the DNA methylation in HCC tissue and plasma, based on MSP method. DNA methylation in plasma, as an non-invasive method, could be used to diagnose HCC, especially for the patients whose AFP is negative. HBV infection may be only associate with DNA methylation of part gene.
ObjectiveTo introduce the new nomenclature scheme of the International Working Group (1995) on hepatic nodules, and summarize the imaging features of various hepatic nodules in light of their pathological characteristics, and evaluate the diagnostic values of various imaging facilities.MethodsUltrasound, computed tomography(CT), magnetic resonance imaging(MRI), and angiographic CT were reviewed and introduced.ResultsMany of these types of hepatic nodules play a role in the de novo and stepwise carcinogenesis of hepatocellular carcinoma(HCC) in the following steps: regenerative nodule, lowgrade dysplastic nodule, highgrade dysplastic nodule, small HCC, and large HCC. Accompanying such transformations, there are significant alterations in the blood supply and perfusion of these hepatic nodules.ConclusionModern stateoftheart medical imaging facilities can not only delineate and depict these hepatic nodules, but also provide important clues for the characterization of focal hepatic lesions in most cases, thus facilitating the early detection, diagnosis and management of HCC in its early stage.
Objective To investigate the expressions of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma, and the correlations of each other in the development of carcinoma. Methods The expressions of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma were detected by S-P immunohistochemical staining, 20 cases of normal liver tissues were collected as contrast, and to compare the relations between expression and prognosis or survival rate. Results The positive rate of CD90, IGF1R, and hTERT protein in hepatocellular carcinoma group were obviously higher than that in contrast group（P＜0.05）, which was 63.9% vs. 0, 52.8% vs.5.0%, and 47.2% vs.0, respectively. The positive rate of CD90, IGF1R, and hTERT protein were higher in UICC Ⅲ-Ⅳ stage group than that in UICC stage Ⅰ-Ⅱ group（P＜0.05）, which was 79.2% vs.33.3%, 70.8% vs.16.7%, and 62.5% vs.16.7%, respectively. There was a statistically significant positive correlation observed between the expressions of CD90 and IGF1R protein （Kendall’s tau-b=0.563 1, P＜0.05）, so it was with CD90 and hTERT protein （Kendall’s tau-b=0.363 6, P＜0.05）. The survival rates of positive expressions of CD90, IGF1R, and hTERT protein were lower than negative expressions of CD90, IGF1R, and hTERT（P＜0.05）, which was 21.7% vs.50.0%, 17.6% vs.43.8%, and 20.0% vs.38.9%, respectively. Conclusions The expressions of CD90, IGF1R, and hTERT may have correlations with the progress of HCC, and may serve as a marker for HCC prognosis potentially.
【Abstract】Objective To investigate the expression of the mRNA of cancer-testis antigen 9 (CT9) gene in hepatocellular carcinoma. Methods The expression of CT9 mRNA was detected through RT-PCR in HCC tissues and their adjacent non-HCC tissues from 45 HCC patients. From CT9 RT-PCR positive products, 3 samples were selected randomly and were sequenced. ResultsCT9 mRNA was detectable in 51.1%(23/45) of HCC samples, and no expression of CT9 mRNA was detected in the adjacent non-HCC tissues. In addition, the RTPCR products were proved to be CT9 cDNA by DNA sequencing. No relationship was found between the expression of CT9 mRNA and clinical factors such as age, sex, tumor size, degree of tumor differentiation, serum αfetoprotein level and infection of hepatitis B virus or hepatitis C virus （Pgt;0.05）. ConclusionCT9 mRNA is expressed with high percentage and specificity in hepatocellular carcinomas. The CT9 gene product is a potential target for antigenspecific immunotherapy of HCC.
Objective To investigate the relationship between syndecan-1 protein and malignant phenotypes of hepatocellular carcinoma (HCC). Methods forty-seven formalin-fixed sections were obtained. The syndecan-1 was measured with immunohistochemistry assay (ABC method). Results Among 47 HCC tissues, 28 (59.6%) show negative staining, the syndecan-1 negative rate in HCC with poor differentiation was higher than those with good differentiation (78.3% vs 41.7%, Ｐ＜0.05), and negative rate in large HCC was higher than in small HCC (81.0% vs 42.3%, Ｐ＜0.01), and negative rate in HCC with the presence of tumor-cells in blood was greater than in those without tumor-cells in blood (84.2% vs 42.9%, P＜0.01). No correlation was found between syndecan-1 expression and serum AFP level and Child class. Conclusion Syndecan-1 expression is correlated with the growth, differentiation, invasiveness, metastasis and progression of HCC. It is possible that syndecan-1 is a negative regulator of these malignant phenotypes of HCC, can be regarded as suppressive gene.
Objective To study the expression of proapoptosis gene bax in hepatocellular carcinoma (HCC) and its clinical significance.Methods The protein expression levels of bax were determined by immunohistochemistry with Envision system; bax mRNA was detected by in situ hybridizition. Results Using immunohistochemistry and in situ hybridizition method, bax protein was detectable in 47.50% of HCC and in 78.57% of cirrhosis (Ｐ＜0.05); there was significant relationship between bax expression and grades of differentiation, between bax expression and clinical stages, and between bax expression and AFP levels (Ｐ＜0.05, Ｐ＜0.01, respectively). However, they were no statistical difference between the male and female or the old and young patient (Ｐ＞0.05, respectively). The expression of bax mRNA by in situ hybridizition were corresponding to immunohistochemistry, and there were no statistical difference between them. Conclusion Proapoptotic gene bax participates partially apoptosis regulation in HCC, the expressions show some correlation with grades, clinical stages and levels of AFP.