Role of R-Spondin1 in the activation of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the role of R-Spondinl in the activation of hepatic stellate cells (HSCs).</p><p><b>METHODS</b>Twenty-four healthy male Kunming mice were randomly divided into the following two groups:fibrosis model group (n=16) and control group (n=8). Hepatic fibrosis was induced by subcutaneous injections of CC14 (20% in olive oil) at a dose of 5 ml/kg twice per week. After 10 weeks, the animals were sacrificed by CO(2) over-exposure and liver tissues were harvested.The protein and mRNA levels of R-Spondin1, alphat-SMA,and collagen I were examined by Western blot assay and real-time PCR respectively. Additionally,HSCs were isolated from the mice liver tissues to examine the time-series expression changes of R-Spondinl, alpha-SMA, and nuclear beta-catenin.TCF activity was analyzed by luciferase reporter assay.Moreover,HSCs were cocultured with recombinant R-Spondin1 and DKK1 to evaluate dose-response.</p><p><b>RESULTS</b>R-Spondinl expression was significantly higher in the fibrosis model group than in the control group (protein level:3.16 ± 0.18 vs. 0.99 ± 0.16, t =13.31, P < 0.01; mRNA level:4.36 ± 0.26 vs. 0.98 ± 0.12, t =21.46, P < 0.01).The culture-activated mouse HSCs showed up-regulated TCF activity (5.33 ± 0.34 vs. non-activated: 1.03 ± 0.09, t =20.93, P < 0.01), nuclear beta-catenin expression (4.47 ± 0.21 vs. 0.97 ± 0.14, t =25.25, P < 0.01), and R-Spondin1 expression (protein level: 4.54 ± 0.18 vs. 1.04 ± 0.12, t =31.17, P < 0.01; mRNA level:5.13 ± 0.15 vs. 1.01 ± 0.16, t=38.06, P < 0.01). Exogenous stimulation of freshly isolated mouse HSCs with recombinant R-Spondin1 induced a dose-dependent increase in both TCF activity and the expression of nuclear beta-catenin and alphat-SMA. DKK1 down-regulated activities of factors in the WNT signaling pathway and repressed activation of HSCs. Conclusion R-Spondin1 may promote HSC activation by enhancing the canonical WNT signaling pathway.</p>

The role of Hedgehog signaling pathway effective proteins in hepatocellular carcinoma / 中华肝胆外科杂志

Objective This study analyzes the expression and clinical significance of Gli1 and Gli3 proteins in hepatocellular carcinoma.Methods 36 patients with hepatocellular carcinoma were studied.The expressions of Gli1 and Gli3 in the carcinoma and adjacent non-tumor tissues were detected with immunohistochemical assay,and their correlations with clinicopathological factors were statistically analyzed.Results Expression rates of Gli1 in hepatocellular carcinoma and adjacent nontumor tissues were 75 ％ and 36.1％,respectively.Expression rates of Gli3 in hepatocellular carcinoma and adjacent non-tumor tissues were 58.3％ and 30.6％,respectively.Expression rates of Gli1 and Gli3 in hepatocellular carcinoma were significantly higher than in adjacent non-tumor tissues (P＜0.05),and a positive correlation was found between the expression of Gli1 and Gli3 (r=0.423,P＜0.05).There was no association between the expression of Gli3 and clinicopathological factors such as age,tumor size,tumor differentiation and lymphatic metastasis.The expression of Gll1 was not related witha patient's age and tumor size,hut there were significant associations with tumor differentiation and lymphatic metastasis.Conclusions Therefore,the expression rate of Gli1 was positively correlated with tumor malignancy,which makes the detection of Gli1 and Gli3 valuable for the diagnosis and prognosis of hepatocellular carcinoma.