Jikai Yin

Clinicopathological significance of SOX4 expression in primary gallbladder carcinoma

AimSOX4, as a member of the SRY-related HMG-box (SOX) transcription factor family, has been demonstrated to be involved in tumorigenesis of many human malignancies; however, its role in primary gallbladder carcinoma (PGC) is still largely unknown. The aim of this study was to investigate SOX4 expression in PGC and its prognostic significance.MethodsFrom 1997 to 2006, 136 patients underwent resection for PGC. The median follow-up was 12.8 months. Immunostainings for SOX4 were performed on these archival tissues. The correlation of SOX4 expression with clinicopathological features including survival was analyzed.ResultsSOX4 was expressed in 75.0% (102/136) of PGC but not in the normal epithelium of the gallbladder. In addition, the over-expression of SOX4 was significantly associated with low histologic grade (P = 0.02), low pathologic T stage (P = 0.02), and early clinical stage (P = 0.03). The levels of SOX4 immunostainings in PGC tissues with positive nodal metastasis were also significantly lower than those without (P = 0.01). Moreover, Kaplan-Meier curves showed that SOX4 over-expression was significantly related to better overall (P = 0.008) and disease-free survival (P = 0.01). Furthermore, multivariate analyses showed that SOX4 expression was an independent risk factor for both overall (P = 0.03, hazard ratio, 3.682) and disease-free survival (P = 0.04, hazard ratio, 2.215).ConclusionOur data indicate for the first time that the over-expression of SOX4 in PGC was significantly correlated with favorable clinicopathologic features and was an independent prognostic factor for better overall and disease-free survival in patients. Therefore, SOX4 might be an auxiliary parameter for predicting malignant behavior for PGC.Virtual slidesThe virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1534825818694957.

Downregulation of microRNA-139 is associated with hepatocellular carcinoma risk and short-term survival

MicroRNAs (miRNAs) have been frequently reported to be diagnostic biomarkers and prognostic factors for cancer. The aim of the present study was to evaluate the clinical significance of microRNA-139 (miRNA-139) in hepatocellular carcinoma (HCC). All 31 patients enrolled in the present study had received curative hepatectomy. The objective miRNA was determined using miRNA microarray. The miRNA-139 expression level in cancerous tissue specimens was measured by means of reverse transcription and quantitative polymerase chain reaction, and compared with that in 31 corresponding peritumoral non-cancerous tissues. Plasma miRNA-139 expression was also quantified. The diagnostic value of plasma miRNA-139 for differentiating patients with HCC from the ones with chronic HBV-hepatitis (CH) was analyzed. The miRNA microarray performed in 3 pairs of tissue specimens determined miRNA-139 was downregulated (p=0.017). Compared with plasma of chronic HBV-hepatitis, miRNA-139 was lowly expressed in plasma of HCC patients (p<0.010). ROC analysis of plasma miRNA-139 yielded an AUC of 0.764 (p<0.010) with sensitivity of 80.6% and specificity of 58.1% while differentiating HCC from chronic HBV-hepatitis. The diagnostic power of serum α-fetoprotein (AFP) was also evaluated. The combination of miRNA-139 and AFP improved the differentiating power. Subsequently, 31 HCC patients were divided into the low or high expression group based on plasma miRNA-139 level. Plasma miRNA‑139 expression was correlated with serum AFP (p=0.043), Edmondson-Steiner grading (p=0.038). In addition, there was a significant difference in the 1-year survival rates between the two groups (p=0.023). miRNA‑139 was downregulated in the cancerous tissue and plasma of HCC patients. Plasma miRNA‑139 is a diagnostic biomarker and prognostic factor for HCC.

Upregulated circulating miR-150 is associated with the risk of intrahepatic cholangiocarcinoma

During the last decade, microRNAs (miRNAs) have been identified as potential biomarkers and therapeutic targets for multiple malignancies; yet, few studies exist on intrahepatic cholangiocarcinoma (ICC). In the present study, a miRNA microarray was applied to determine the significant miRNAs involved in ICC. miR-150 was found to be significantly downregulated in ICC. We further enrolled 15 ICC patients who received radical resection to test these findings in plasma. Using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), we examined and quantified the expression levels of miR-150 in tumor tissues, peritumoral noncancerous tissues and blood samples of 15 ICC patients. The diagnostic value of plasma miR-150 for differentiating patients with ICC from the age- and gender-matched controls was analyzed. For plasma samples, compared with normal controls, the level of miR-150 expression was found to be upregulated (P<0.010) in ICC patients. While differentiating ICC from normal controls, receiver operator curve (ROC) analysis of plasma miR-150 revealed the area under the curve (AUC) of 0.764 (P<0.010) with sensitivity of 80.6% and specificity of 58.1%. The diagnostic value of carbohydrate antigen 19-9 (CA19-9) and the combination of miR-150 and CA19-9 were also evaluated. We found that the combination of these two markers improved the power of screening ICC. Moreover, on the basis of the plasma miR-150 level, 15 ICC patients were divided into a low or high expression group. We found that plasma miR-150 is a potential diagnostic biomarker for ICC.

Inhibition of Janus kinase-2 signalling pathway ameliorates portal hypertensive syndrome in partial portal hypertensive and liver cirrhosis rats

BACKGROUND AND AIMS JAK2/STAT3 signalling promotes fibrosis, angiogenesis and inflammation in many diseases; however, the role of this pathway in portal hypertension remains obscure. This study aimed to explore the function of JAK2/STAT3 signalling in portal hypertension and estimate the potential therapeutic effect of treatment with the specific inhibitor AG490. METHODS Rats induced by partial portal vein ligation and common bile duct ligation were treated with AG490 for two weeks. Haemodynamic parameters were assessed. The levels of phospho-STAT3 protein and related cytokines were detected by western blotting of splanchnic organs. Liver, spleen and intestine characterization was performed using histological analyses. Peripheral blood cell counts were also detected. RESULTS High levels of phospho-STAT3 protein were detected in portal hypertensive rats. AG490 effectively inhibited JAK2/STAT3 signalling and its downstream cytokines and provided protective effects by decreasing splanchnic neovascularization and inflammation and by attenuating portal pressure and hyperdynamic splanchnic circulation. In cirrhosis rats, AG490 inhibited intrahepatic fibrosis, angiogenesis and inflammation. AG490 improved the peripheral blood cell counts and the splenomegaly observed in these rats. CONCLUSIONS JAK2/STAT3 signalling is essential in portal hypertension, and targeting JAK2/STAT3 may be a promising therapy to treat this condition.

Two surgical procedures for esophagogastric variceal bleeding in patients with portal hypertension.

AIM To determine the clinical value of a splenorenal shunt plus pericardial devascularization (PCVD) in portal hypertension (PHT) patients with variceal bleeding. METHODS From January 2008 to November 2012, 290 patients with cirrhotic portal hypertension were treated surgically in our department for the prevention of gastroesophageal variceal bleeding: 207 patients received a routine PCVD procedure (PCVD group), and 83 patients received a PCVD plus a splenorenal shunt procedure (combined group). Changes in hemodynamic parameters, rebleeding, encephalopathy, portal vein thrombosis, and mortality were analyzed. RESULTS The free portal pressure decreased to 21.43 ± 4.35 mmHg in the combined group compared with 24.61 ± 5.42 mmHg in the PCVD group (P < 0.05). The changes in hemodynamic parameters were more significant in the combined group (P < 0.05). The long-term rebleeding rate was 7.22% in the combined group, which was lower than that in the PCVD group (14.93%), (P < 0.05). CONCLUSION Devascularization plus splenorenal shunt is an effective and safe strategy to control esophagogastric variceal bleeding in PHT. It should be recommended as a first-line treatment for preventing bleeding in PHT patients when surgical interventions are considered.

Targeted sequencing of cancer-associated genes in hepatocellular carcinoma using next generation sequencing.

Liver cancer is one of the most common causes of cancer-associated mortality. Hepatocellular carcinoma (HCC) is the major histological subtype among types of primary liver cancer. China is an area of high incidence of HCC, and >50% of the cases of HCC worldwide are in China. At present, the mechanism underlying the development of HCC remains to be fully elucidated, and previous studies have predominantly focused on HCC in southern and eastern China, with molecular data of the HCC cases in Western China remains limited. In the present study, a panel of 372 cancer‑associated genes were screened using a next generation sequencing platform, which included a total of 12 cases from western China. The results confirmed mutations in previously identified HCC drivers, including p53 and Kras. Additionally, mutations in several cancer genes, which had not been previously associated with HCC, were identified, including RUNX1 and JAK3. The present study provided a mutation spectrum of HCC tissue in cases from western China, assisting in the investigation of the mechanism of liver carcinogenesis.

Combined administration of propranolol + AG490 offers better effects on portal hypertensive rats with cirrhosis.

AG490, the specific inhibitor of JAK2/STAT3 signaling, has been shown to decrease portal pressure, splanchnic hyperdynamic circulation and liver fibrosis in cirrhotic rats. Nonselective betablockers such as propranolol are the only drugs recommended in the treatment of portal hypertension. The aim of this study was to explore the combinative effect of treatment with propranolol and AG490 on portal hypertension.

A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro.

The growth factor receptor bound protein 2 (Grb2) is one of the affirmative targets for cancer therapy, especially for breast cancer. In this study, we hypothesized the Src-homology 2 (SH2) domain in Grb2 may serve as a competitive protein-binding agent to interfere with the proliferation of breast cancer cells in vitro. We designed, constructed, expressed and purified a novel fusion protein containing the protein transduction domain (PTD) and Grb2-SH2 domain (we named it after PTD-Grb2-SH2). An immunofluorescence assay was used to investigate the location of PTD-Grb2-SH2 in cells. MTT assay and EdU experiments were applied to detect the proliferation of breast cancer cells. The ultra-structure was observed using transmission electron microscopy. Flow cytometry was used to determine the cytotoxicity of PTD-Grb2-SH2 on cell proliferation. We successfully obtained the PTD-Grb2-SH2 fusion protein in soluble form using a prokaryotic expression system. The new fusion protein successfully passed through both the cellular and nuclear membranes of breast cancer cells. The MTT assay showed that PTD-Grb2-SH2 exhibited significant toxicity to breast cancer cells in a dose- and time-dependent manner in vitro. EdU identified the decreased proliferation rates in treated MDA-MB-231 and SK-BR-3 cells. Observation by transmission electron microscopy and flow cytometry further confirmed the cytotoxicity as apoptosis. Our results show that the HIV1-TAT domain is a useful tool for transporting a low molecular weight protein across the cell membrane in vitro. The PTD-Grb2-SH2 may be a novel agent for breast cancer therapy.

Effect of mycophenolate mofetil plus adriamycin on HepG-2 cells.

AIM To investigate the influence of mycophenolate mofetil (MMF) plus adriamycin (ADM) on hepatocellular carcinoma (HCC) cells. METHODS HCC cells were treated with 100 μg/ml of MMF alone (MMF group), 1 μg/mL of adriamycin (ADM group) alone, or a combination of the drugs (MMF + ADM group). We performed an 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay to measure the growth inhibition rate of HCC cells. Flow cytometry was used to determine the percentage of cells in different phases of the cell cycle and the number of apoptotic cells. Hoechst 33258 staining revealed the morphological changes associated with apoptosis in HCC cells. RESULTS The results of MTT assays revealed that monotherapy with ADM or MMF showed inhibition of cell growth, while MMF + ADM therapy afforded an inhibition rate of more than 90% with cell distribution in G1 and G2/M phase greater than that in S phase. MMF + ADM treatment also downregulated Bcl-2 expression markedly. The growth of HCC cells was markedly inhibited and apoptosis was enhanced in all the 3 groups. Compared with other 2 groups, the MMF + ADM group showed more obvious apoptosis of cells. CONCLUSION The MMF plus ADM combination exerts remarkable inhibitory effects on the growth of HCC cells.