Herpes simplex virus-thymidine kinase gene transduced into T cell clone in the treatment of ulcerative colitis in rat / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of herpes simplex virus-thymidine kinase (HSV-TK) gene transduced into T cell clone on the treatment of ulcerative colitis (UC) in rat.</p><p><b>METHODS</b>The T cell clone transduced with HSV-tk was infused into 27 rats with UC. Changes of stimulation index (SI), CD4(+), CD8(+), IL-13, IL-4 were detected, and pathological changes before and after infusion was compared.</p><p><b>RESULTS</b>In the second and third day after tk+ T clone infusion, the inflammation of colon was assimilated. Two weeks later, the colon began to renovate and mend. The average SI was 7.39+/-1.24 before infusion, and 2.67+/-0.87 after infusion (P<0.05). Peripheral blood levels of CD4(+), CD8(+) or IL-13 and IL-4 in therapeutic group were significantly decreased as compared to control group (P<0.05).</p><p><b>CONCLUSION</b>HSV-tk gene transduced into T lymphocyte clone and infused back is effective for UC gene therapy and target design in rat.</p>

Experimental research of therapeutic effect on hepatocellular carcinoma of targeting SMYD3 gene inhibition by RNA interference / 中华外科杂志

<p><b>OBJECTIVE</b>To determine the potential of SMYD3 as a therapeutic target for hepatocellular carcinoma (HCC) by potent and highly sequence-specific RNA interference (RNAi) technique.</p><p><b>METHODS</b>The mRNA of SMYD3 was detected by RT-PCR in different HCC cell lines, such as HepG2, Hep3B and SMMC7721. Recombinant SMYD3 shRNA plasmid Pgenesil-1-s was constructed and transfected into HepG2 cells, and Western blot was used to identify the down regulation of SMYD3 protein expression after transfection. MTT and flow cytometry analysis (FCM) were respectively applied to analysis cell proliferation and apoptosis. In vivo study was carried out by injecting recombinant SMYD3 shRNA plasmids into transplanted tumors of nude mice.</p><p><b>RESULTS</b>The expression of SMYD3 mRNA was abundant in HCC cell lines HepG2, Hep3B, SMMC7721, whereas none in normal hepatic cell line L-02. RNA interference was able to suppress SMYD3 expression greatly and then inhibited cell growth effectively and induced apoptosis of HepG2 cells efficiently. After injection of recombinant SMYD3 shRNA plasmid, transplanted tumors grew slowly and reduced in size and weight when compared with those of control groups (P < 0.01).</p><p><b>CONCLUSIONS</b>SMYD3 plays a major role in occurrence and progress of HCC. Inhibition of SMYD3 by RNAi can induce apoptosis in HepG2 cells and suppress tumor growth in nude mice. Therefore SMYD3 could be an ideal therapeutic target for HCC.</p>

Suppression of SMYD3 expression in HepG2 cell by shRNA interference / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To identify the inhibition effect of shRNA on the SMYD3 (SET- and MYND-domain containing protein-3) expression in hepatoma cell line HepG2 through gene silencing.</p><p><b>METHODS</b>Two reverse repeated motifs targeting on the SMYD3 mRNA sequences 267-288, 302-323 respectively, were synthesized and inserted into the mock plasmid pGenesil-1 which expressed EGFP to create recombinant plasmids pGenesil-1-s1 and pGenesil-1-s2. pGenesil-1-hk specific to no SMYD3 mRNA sequence served as a control. After transfection into HepG2 cells, RT-PCR and western blot were applied to identify the down regulation of SMYD3 expression by shRNAs.</p><p><b>RESULTS</b>All plasmids were constructed successfully. pGenesil-1-s1, pGenesil-1-s2 inhibited the mRNA and protein expression of SMYD3 in HepG2 cells. There was a significant distinction when compared with pGenesil-1-hk and pGenesil-1 (P<0.01).</p><p><b>CONCLUSION</b>Short hairpin RNAs can efficiently and specifically suppress the expression of SMYD3 in HepG2 cells.</p>