Reconstructing the JAK/STATs signal pathway restored the anti-proliferative response of MHCC97 on interferon alpha / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To elucidate the roles of JAK/STATs signal pathway on anti-proliferative effects induced by IFN-alpha in MHCC97.</p><p><b>METHODS</b>An IRF9 expression vector was transfected into MHCC97 with Dosper. The expression of IRF9, cycle regulating proteins and the forming of ISGF3 complex were detected using Western blot and EMSA, respectively. Cell proliferation and distribution were monitored using MTT and flow cytometry.</p><p><b>RESULTS</b>High expression of IRF9 restored the anti-proliferative response of MHCC97 on IFN-alpha treatment and delayed the cell transition from S phase to G2 phase induced by IFN-alpha.</p><p><b>CONCLUSION</b>The integrity and functions of JAK/STATs signal pathway played an important role in mediating the anti-proliferative effects of IFN-alpha in MHCC97.</p>

The effects of ezrin on the proliferation and the invasiveness of cells of different hepatocellular carcinoma cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the different expressions of cytoskeletal organizer ezrin and cytoskeleton protein beta- and gamma-actin in hepatocellular carcinoma (HCC) cell lines with different metastatic potentials and to explore the role of ezrin in cell growth and metastasis in HCC cell lines SF7721 and MHCC97-H.</p><p><b>METHODS</b>Immunofluorescence, RT-PCR and Western blot were used to detect the gene and protein expressions of ezrin and actin in hepatocellular carcinoma cell lines with different metastatic potentials. RNA interference (RNAi) was applied to down-regulate the ezrin expression in SF7721 and MHCC97-H. Changes of the cell growth and metastasis potentials after the RNAi treatment were studied. MTT assay was used to detect cell proliferation changes and Transwell assay was applied to observe the changes of cell motility and invasiveness.</p><p><b>RESULTS</b>Both ezrin and cytoskeleton protein were demonstrated in the cytoplasma of the cells at the same time. The expression of them in cell lines with high metastatic potential, such as SF7721, MHCC-1 and MHCC97-H was obviously higher than in those with low metastatic potentials, such as SMMC-7721, Hep3B and HepG2 (chi2= 13.277, P = 0.010; chi2= 21.815). The mRNA and ezrin and cytoskeleton protein gamma-actin were over-expressed in HCC cell lines with high metastatic potentials. The expressions of beta-actin of cell lines with different metastatic potentials showed no differences. Ezrin protein was successfully down-regulated and the proliferation and the invasiveness of the cells decreased with low ezrin protein level in SF7721 and MHCC97-H.</p><p><b>CONCLUSION</b>Over-expression of ezrin and cytoskeleton protein gamma-actin are associated with the process of metastasis of hepatocellular carcinoma cells. The growth and invasiveness of SF7721 and MHCC97-H cells can be inhibited by down-regulating ezrin expression.</p>

Influence of the inhibitor of c-Met on the growth and motility of hepatocellular carcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To explore the influence of c-Met inhibitor by synthetic c-Met antisense oligonucleotide, constructive c-Met antisense plasmid and the complex plasmid of U1SnRNA/ ribozyme/anti-Met on the growth and metastasis of hepatocellular carcinoma cells.</p><p><b>METHODS</b>Gene transfection was operated by Lipofectin on SF7721 cells. The difference of the cells before and after transfection was compared by MTT, growth curves and transwell test in vitro. In vivo, the cells before and after transfection were implanted subcutaneously into nude mice respectively to observe tumor growth and metastasis.</p><p><b>RESULTS</b>C-Met antisense oligonucleotide could inhibit the growth of hepatocellular carcinoma SF7721 cells (t=3.58, P<0.05). After transfection, the expression of c-Met protein decreased. Growth curves showed that the cells after transfection proliferated more slowly, about 50% of control cells (F=4.87, P<0.05), and their motility and invasiveness decreased, compared with those before transfected. In vivo experiment, tumors originated from c-Met antisense oligonucleotide treated cells and the antisense/ribozyme/U1SnRNA treated cells grew more slowly (about 54.5% of those from the control cells), and the latent prolonged. After 35 days, the average weight of tumors in the two group nude mice were lighter than that in the control group nude mice (F=5.17, P<0.05).</p><p><b>CONCLUSION</b>Inhibition of c-Met expression by c-Met antisense oligonucleotide and the complex of antisense/ribozyme/U1SnRNA can inhibit the growth and metastasis of SF7721 hepatocarcinoma cells in vitro and in vivo.</p>