Selective blockage of abnormal Wnt pathway inhibits the growth and invasion of hepatocarcinoma cells / 复旦学报（医学版）

Objective To construct the recombinant expression vector encoding antisense Tcf fragment for the blockage of abnormal Wnt pathway, and to investigate its effect on the biological behaviors of human hepatocarcinoma cells. Methods Antisense expression vector was transfected into hepatocarcinoma cells SMMC-7721 with GeneJammer. RT-PCR and Western blot were used to detect Tcf expression. Cell proliferation and motility were compared by growth curves and Transwell plate assay. Cell apoptosis was determined by Annexin V and cell cycle was examined by fluorescent staining. Results The stable transfection of antisense Tcf in SMMC-7721 cells significantly reduced Tcf expression at both mRNA and protein levels. Compared with parental and mock-transfected 7721 (7721-vector) cells, antisense Tcf RNA transfected cells 7721-pTas showed much decreased activities of proliferation, migration and invasion in vitro. Furthermore, the apoptosis rate of 7721-pTas cells [(26.34±2.07)%] was significantly higher than that of 7721-vector cells [(6.53±1.02)%] and parental SMMC-7721 cells [(4.33±0.68)%] (P<0.001). The percentages of G0-G1 phase antisense transfected cells were 20.24% and 20.95%, higher than parental SMMC-7721 and 7721-vector cells, and percentages of S phase antisense transfected cells were 11.8% and 11.38%, lower than parental SMMC-7721 and 7721-vector cells, respectively. Conclusions Antisense RNA suppress the growth ability of liver cancer cells by inducing cell apoptosis and impeding the progress of cell cycle, which suggests that selective blockage of abnormal Wnt signal pathway by antisense Tcf RNA may be a potential new gene therapy for liver cancer.

Surgical treatment of primary hepatoeellular carcinoma: a 20-year clinical experience in 7566 patients / 中华消化外科杂志

Objective To summarize the clinical experienee in surgical treatment for hepatocellular carcinoma (HCC). Methods The clinical data of 7566 HCC patients who had been admitted to Research Institute of Liver Cancer of Fudan University from January 1988 to Deeember 2007 were retrospectively analyzed. The overall survival and recurrence free survival (RFS) rates were eaeulated with Kaplan-Meier survival curve. All the data were analyzed using Log-rank test and Cox regression model. Results The 3-, 5-, 10-year overall survival and RFS rates of 7164 patients with HCC resection were 56.29%, 41.76%, 26.70%, and 63.92%, 56.12%, 42.97%, respectively, and the perioperative mortality was 1.54%. The 5- and 10-year overall survival rates of patients with small HCC (diameter<5 era) were 58.20% and 38.47%, which were significantly higher than 31.42% and 20.43% of patients with large HCC (diameter >5 cm) (X2 =535. 568, P <0.01). The 5-year overall survival rotes of HCC patients with resection after down-staging (n = 110), re-resection after recurrence (n = 515), and tumor thrombus in portal vein (n = 168) were 51.26%, 67.28% and 26.81%, respectively; nd the 5-year DFS rotes were 77.44%, 13.01% (calculated from the first operation) and 34.90%, respectively. The 3- and 5-year overall survival and DFS rates of 402 patients who had undergone liver transplantation were 60.81%, 55.63% and 64.47%, 58.52%. The independent prognostic factors influencing the overall survival and DFS rates were the size, number and differentiation of HCC and intrahepatic vessel invasion (X2 = 200.539, 27. 536, 96.964,216. 156, P <0.01). Conclusions Early screening, improved safety of surgery, combined therapy and breakthrough in the reseaeh of preventing HCC metastasis and reeurrenee will significantly improve the treatment outcome of HCC.

Alterations of beta-catenin and Tcf-4 instead of GSK-3beta contribute to activation of Wnt pathway in hepatocellular carcinoma / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>The goal of this study is to investigate the inappropriate activation of Wnt pathway in the hepatocarcinogenesis.</p><p><b>METHODS</b>We analyzed the alterations of three key components of Wnt pathway, beta-catenin, glycogen synthase kinase 3beta (GSK-3beta) and T cell factor 4 (Tcf-4), in 34 samples of hepatocellular carcinoma (HCC) and paracancerous normal liver by immunohistochemistry, polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), direct sequencing, semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization.</p><p><b>RESULTS</b>We found 61.8% (21/34) of all the HCCs examined showed an abnormal beta-catenin protein accumulation in the cytoplasm or nuclei. RT-PCR-SSCP and direct sequencing showed that beta-catenin exon 3 mutations existed in 44.1% (15/34) of the HCCs. No mutations of GSK-3beta or Tcf-4 were detected in HCCs. Moreover, mRNA of beta-catenin and Tcf-4 but not GSK-3beta was found to be over expressed in HCCs. On analyzing the relationship between alterations of beta-catenin or Tcf-4 and C-myc or Cyclin D1 expression, we found that the mutations of beta-catenin as well as over expression of beta-catenin or Tcf-4 gene were independently correlated with C-myc gene over expression in HCCs.</p><p><b>CONCLUSIONS</b>Our present findings strongly suggest mutations of beta-catenin as well as over expression of beta-catenin and Tcf-4 gene activate the Wnt pathway in HCC independently with the target gene most likely to be C-myc.</p>

Focal nodular hyperplasia of the liver: a report of 60 cases / 中国普通外科杂志

Objective To evaluate the diagnosis and treatment of focal nodular!hyperplasia of the liver (FNH). Methods Retrospective analysis was made on 60 FNH cases in terms of clinical findings, images, pathologic examination and surgical treatment. Results Of the 60 FNH patients in our hospital from 1993 to 2003, 41 were male and 19 female. The average age was 37 year′s old. Fifty-five cases had single focus, the other five were of multiple lesion, with tumor diameter 10cm in one. Correct preoperative diagnosis was made in 33 cases (55%). The correct diagnostic rate of BUS, CT and MRI was 33.3%, 58.3% and 72.0%, respectively. All 60 cases underwent operation with an uneventful recovery and without recurrence at follow-up. ConclusionsCT and MRI are mandatory for the diagnosis of FNH. Definite preoperative diagnosis is usually difficult even in cases of typical type of FNH. Surgical resection is the treatment of choice when a patient becomes symptomatic or when malignancy could not be excluded.