Relative analysis of OPN and its related signal molecules in hepatocellular carcinoma / 中华肝脏病杂志

Osteopontin (OPN) has close relationship with metastasis in hepatocellular carcinoma but its downstream signal pathways have not been well defined in hepatocellular carcinoma. The object of this study is to identify the associated signal pathways in human HCC tissues. The expressions of OPN, intergrin aV, CD44v6, P-FAK, FAK, P-Src, Src, P-ERK and P-AKT were assayed using TMA analysis. The relationship of OPN with P-ERK, P-Src and P-AKT were explored and the role in HCC metastasis was analysed. The expression levels of OPN, intergrin aV, CD44v6, P-FAK, P-Src, Src, P-ERK and P-AKT in HCC tissue were significantly higher than that in normal tissue (P value is less than 0.05). No significant difference was found between the expression levels of FAK in HCC tissue and normal tissue (P value is more than 0.05). OPN expression was significantly associated with Integrin av (P value is less than 0.01), CD44V6 (P value is less than 0.01) and P-ERK (P value is less than 0.05) but not with P-Src, P-FAK and P-AKT (P value is more than 0.05). The expressions of P-FAK (P value is less than 0.05), P-Src (P value is less than 0.01) and P-AKT (P value is less than 0.05) were significantly associated with Integrin av and the P-FAK expression was also significantly associated with CD44V6 (P value is less than 0.01). OPN promotes HCC metastasis though Integrin av/CD44V6/MAPK pathway in human HCC.

Alteration of the malignant phenotype of human hepatocellular carcinoma cell line Hep3B by specially synthesized oligodeoxynucleotides / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the inhibitory effect of specially synthesized oligodeoxynucleotide (SODN) on malignant phenotype of human hepatocellular carcinoma Hep3B cells by complementary binding to the fourth promoter of IGF-2 gene.</p><p><b>METHODS</b>A SODN was synthesized according to the sequence of the fourth promoter of IGF-2 gene, and was then transfected into Hep3B cells which overexpressed IGF-2. IGF-2 gene transcription activity and protein expression were assayed by RT-PCR and Western blot methods. The effect on cell growth by SODN was estimated by MTT method, and the effect on cell cycle distribution was measured by flow cytometry. Colony formation assay was performed on 6-well tissue culture dishes. Alteration on mobility and invasiveness were studied used transwell plates.</p><p><b>RESULTS</b>IGF-2 mRNA and protein levels in Hep3B cells transfected with SODN were significantly lower in comparison with those in control groups (Hep3B cells and Hep3B cells transfected with a control oligodeoxynucleotide). Results also showed that SODN did not have inhibitory effects on cell growth and mobility of Hep3B cells, but did have an effect on its colony formation and invasiveness.</p><p><b>CONCLUSION</b>SODN has inhibitory effect on IGF-2 expression in Hep3B cells as a molecular switch, which partially alterates the malignant phenotype of this cell line.</p>

Transcatheter hepatic arterial chemoembolization on recurrent hepatocellular carcinoma after resection / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effect of transcatheter hepatic arterial chemoembolization (TACE) therapy on the survival and prognosis of recurrent hepatocellular carcinoma (HCC) after surgical resection.</p><p><b>METHODS</b>The data of 130 surgically resected but recurrent HCC patients treated by TACE were reviewed retrospectively. The survival and influencing factors on the prognosis were analyzed.</p><p><b>RESULTS</b>The overall 1-, 3-, 5-year survival rates of these 130 patients were 83.0%, 45.5% and 17.6% respectively (median survival time 2.4 years). Ninty-four of the series were treated with TACE alone, which gave the 1-, 3- year survival rates of 76.4% and 37.1%, respectively (median survival time 2.1 years). Thirty-six out of 130 patients treated with TACE plus percutaneous ethanol injection (PEI), the 1-, 3-year survival rates were 100.0% and 66.5% respectively with a median survival time (MST) of 3.5 years. The survival of TACE plus PEI group was significantly better, and the mortality risk was significantly lower than that of TACE alone group (P < 0.05). The mortality risk of those with > 5 cm diameter recurrent tumor or with distant metastasis was significantly higher than those with < or = 5 cm diameter tumor or without metastasis (P < 0.05).</p><p><b>CONCLUSION</b>TACE combined with PEI may improve the survival of recurrent HCC patients.</p>

Analysis on the variance of vacA genotypes and their vacuolating toxin activity of Helicobacter pylori isolates in Zhejiang area / 中华流行病学杂志

<p><b>OBJECTIVE</b>To analyze the variance of vacA genotypes and their vacuolating toxin activity of Helicobacter pylori (H. pylori) isolates from patients with peptic ulcer (PU) or chronic gastritis (CG) in Zhejiang area.</p><p><b>METHODS</b>Signal and middle regions of parts of seventy H. pylori strains were sequenced after T-A cloning. Vacuolating toxin activity was detected with cell culture method.</p><p><b>RESULTS</b>Compared with the reported sequences of H. pylori strain 60190 with s1a/m1 genotype, similarities of the signal-region sequences from 6 s1a-type H. pylori isolates were found to be 93.2%-98.3%, and from 1 m1b-type strain was 87.3%. When compared with the corresponding sequences of H. pylori strain 87-203 with m2 genotype, similarities of the mid-region from 4 m2-type isolates and 1 m1b-type isolates were 93.8%-97.6% and 71.7%, respectively. All 5 strains with s1a/m1 type produced vacuolating toxin activity detected by HeLa, RK-13, and SGC-7901 cell lines. Only 12 strains with s1a/m2 type produced cytotoxin in HeLa cells but 65.1% (28/43) and 62.8% (27/43) strains had cytotoxin in RK-13 and SGC-7901 cells. In RK-13 cells, 81.0% (17/21) strains with s1a/m1b produced vacuolating toxin activity.</p><p><b>CONCLUSION</b>Variance of vacA genotypes of local H. pylori isolates mainly lied in mid-region. H. pylori isolates with m2 type produced high cytotoxin in RK-13 and SGC-7901 cells, and low cytotoxin in HeLa cells. Cytotoxin strength of m1b-type strains was between strains with ml and m2 types. Vacuolating toxin activity of strains from PU group seemed obviously higher than that from CG group.</p>

Dominant cagA/vacA genotypes and coinfection frequency of H. pylori in peptic ulcer or chronic gastritis patients in Zhejiang Province and correlations among different genotypes, coinfection and severity of the diseases / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Almost half of the world's population suffer from the Helicobacter pylori (H. pylori) infection, but only some individuals develop gastric diseases with clinical symptoms. One reason for the phenomenon may be the different pathogenicity of infected H. pylori strains. The presence of cytotoxin-associated gene A (cagA) and expression of vacuolating cytotoxin activity encoded by vacuolating cytotoxin gene A (vacA) are considered the two major virulent markers of H. pylori. The aim of this study was to detect dominant cagA/vacA genotypes and coinfection frequency of H. pylori in patients with peptic ulceration (PU) or chronic gastritis (CG), and to determine correlations among different cagA/vacA genotypes, coinfection and severity of the diseases.</p><p><b>METHODS</b>For each of 139 patients in Zhejiang Province who had been diagnosed as PU or CG based on clinical symptoms and gastroscopy, two gastric biopsy specimens (one from antrum and the other from corpus) for H. pylori isolation were taken by two different disinfected biopsy forceps. One hundred and fifty-six H. pylori strains were isolated from both the antrum and corpus biopsy specimens of 78 patients (36 PU and 42 CG). PCRs were performed to detect cagA genes, and signal (s) and middle (m) regions of vacA genes in the H. pylori isolates. The amplified fragments of dominant vacA gene s and m subtypes from representative H. pylori isolates were sequenced after TA cloning. Dominant cagA/vacA genotypes of the H. pylori isolates, coinfection frequency and correlations among the different genotypes, coinfection and severity of the diseases were determined.</p><p><b>RESULTS</b>Of the H. pylori strains isolated from the antrum specimens, 96.2% were cagA gene positive, as were 97.4% of the H. pylori strains isolated from the corpus specimens. Only one s region subtype (s1a) and four m region subtypes m1, m2, m1b and m1b-m2 of vacA gene were found. The proportions of vacA gene subtypes s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 in the 83 strains isolated from the antrum specimens were 7.2%, 61.5%, 30.1% and 1.2%, respectively, while those in the other 84 strains isolated from the corpus specimens were 9.5%, 58.3%, 28.6% and 3.6%, respectively. s1a/m2 (58.3% vs 30.1%, chi(2) = 13.47, P < 0.01) and then s1a/m1b (28.6% vs 9.5%, chi(2) = 9.88, P < 0.01) were the dominant vacA gene subtypes in the H. pylori isolates. The dominant H. pylori genotype was cagA + s1a/m2 (59.0% from antrum specimens and 57.1% from corpus specimens), and followed by cagA + s1a/m1b (28.9% from antrum specimens and 27.4% from corpus specimens). Sixteen of 78 patients (20.5%) were infected with two or three H. pylori strains with different genotypes. However, no statistically significant differences among cagA occurrence, the different vacA subtypes and PU or CG could be found (each P > 0.05). Similarities of the nucleotide sequences from vacA gene s region PCR products of six isolates and from vacA gene m region PCR products of four isolates were 93.2% to 98.3% and 93.8% to 97.6%, respectively, compared to the reported corresponding sequences.</p><p><b>CONCLUSIONS</b>The dominant genotypes of H. pylori in PU or CG patients in Zhejiang area may be cagA + s1a/m2 and cagA + s1a/m1b. Numerous coinfections with different H. pylori strains in PU or CG patients indicate diversity of the infected H. pylori origins. s and m regions of vacA gene from different H. pylori isolates show high nucleotide sequence similarities. cagA gene positive rate, different vacA gene subtypes and coinfection with different H. pylori strains are not closely associated with severity of the diseases.</p>