Influences of the interferon induced transmembrane protein 1 on the proliferation, invasion, and metastasis of the colorectal cancer SW480 cell lines / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Interferon-induced transmembrane protein 1 (IFITM1) has been identified as a molecular marker of the colorectal tumors; however its influences on the biological behaviors of the colorectal cancer cells are currently unknown. We aimed to study the influences of IFITM1 on the proliferation, invasion, and metastasis of the colorectal cancer SW480 cell lines.</p><p><b>METHODS</b>We constructed IFITM1/pEGFP-C3 recombinant plasmids and transfected them into the colorectal cancer SW480 cell lines. IFITM1/pEGFP-C3 recombinant plasmids were identified by means of immunofluorescence, laser confocal scanning microscopy, and reverse transcription polymerase chain reaction. IFITM1/SW480 cells with stable over-expression of IFITM1 were confirmed by G418 screening. The influences of IFITM1 on the proliferation of the SW480 cell lines were investigated by MTT assay and tumor transplantation experiments in nude mice. Cell invasion experiments were performed to determine the invasion capacity of the IFITM1/SW480 cells. Matrix metalloproteinase 2 (MMP-2) and MMP-9 activities were detected by the gelatin zymographic analysis, and MMP-9 expression by the Western blotting analysis.</p><p><b>RESULTS</b>IFITM1/pEGFP-C3 recombinant plasmids were successfully constructed in this study, and the IFITM1/SW480 cells with stable IFITM1 gene over-expression were confirmed by G418 screening. MTT results showed that the proliferation of the IFITM1/SW480 cells was significantly enhanced (P < 0.01). Tumors were harvested from four weeks old mice. Tumor volumes were (1347.00 ± 60.94) mm(3), (1032.40 ± 111.38) mm(3) and (1018.78 ± 28.83) mm(3); and tumor weights were (1522.34 ± 62.76) mg, (1137.78 ± 97.22) mg and (1155.76 ± 133.31) mg for mice inoculated with the IFITM1/SW480 cells, pEGFP-C3/SW480 cells and SW480 cells, respectively. Tumor volumes and weights from mice inoculated with the IFITM1/SW480 cells were significantly increased (P < 0.01). In addition, the numbers of the SW480 cells and IFITM1/SW480 cells that migrated through Matrigel were 448.64 ± 38.09 and 540.45 ± 44.61, respectively; so the invasive ability of the SW480 cells transfected with IFITM1 gene was significantly greater than that of the SW480 cells (P < 0.01). Gelatin zymographic analysis showed that MMP-9 and MMP-2 protein activities in the IFITM1/SW480 cells were significantly enhanced, and Western blotting analysis showed that MMP-9 expression in the IFITM1/SW480 cells was also increased.</p><p><b>CONCLUSION</b>IFITM1 can enhance the proliferation, invasion, and metastasis of the colorectal cancer SW480 cell lines.</p>

125I brachytherapy combined with chemotherapy for patients with inoperable stage Ⅲ non-small cell lung cancer / 中华核医学杂志

Objective To assess the therapeutic effect of radioactive seed 125I brachytherapy combined with GP chemotherapy regimen (gemcitabine 1000 mg/m2, cisplatin 75 mg/m2) for inoperabale stage Ⅲ non-small cell lung cancer (NSCLC). Methods Thirty-nine documented inoperable stage Ⅲ NSCLC patients, enrolled between January 2005 and June 2008 for the study group, were treated with the combination of 125I brachytherapy and GP regimen. The brachytherapy methods were conducted according to TPS and each patient was treated under those patients were treated with standard GP regimen. Chest CT scans were performed every three months post-procedurally, until disease progression or recurrence. The follow-up time was up to twenty four months after treatment. In the control group, equal amount of Ⅲ stage NSCLC patients were treated with standard GP regimen alone. Chi-square test and survival analysis with Kaplan-Meier and Log-rank were used to compare the differences of recent (3 months after therapy)efficiency, survival rate, survival time between two groups. Results The re-cent effective rates of the study group (71.8%, 28/39) and control group (61.5%, 24/39) were not statistical different (χ2=0.93, P>0.05), yet the tumor CR rates in two groups showed significant disparity (χ2=4.48, P0.05). However, singificant differences (χ2=4.07, 4.63,both P<0.05) were found in 2-year survival rate and median survival time, with 41.0% (16/39) vs 23.1% (9/39) and 18.9±2.7 months vs 14.2±0.7 months. Conclusions 125I brachytherapy combined with GP regimen chemotherapy could be an effective treatment method and could improve the tumor CR rate and survival rate for patients with inoperable stage Ⅲ NSCLC.

Clinic study on cool-tip radiofrequency ablation combined with chemotherapy for advanced non-small-cell lung cancer / 肿瘤

Objective: To explore the therapeutic effects of cool-tip radiofrequency ablation (C-RFA) combined with chemotherapy for advanced non-small-cell lung cancer (NSCLC). Methods: Forty-six patients with advanced NSCLC were treated with C-RFA combined with three cycles of chemotherapy (gemcitabine 1 000 mg/m2, on day 1 and day 8 and cisplatin 100 mg/m2 on day 1). The tumor volume and density were evaluated by chest CT enhancing scan 3 months after C-RFA therapy. All the patients were followed up for 12 months. Natural killer cell (NKc) activity and T cell subgroups in peripheral blood were determined by flow cytometry before and at 1 week after C-RFA therapy. Forty-six NSCLC patients who received three-dimensional conformal radiotherapy (3D-CRT) combined with chemotherapy was regarded as control group. The irradiation dosage of 3 D-CRT was 62-70 Gy. The chemotherapeutic regimen was the same as C-RFA group. Results: After three months of C-RFA therapy, the chest CT imaging showed that the areas of tumor lesions were decreased to different extent in majority cases and the tumor density decreased significantly. There were 7cases of complete response (CR), 29 cases of partial response (PR), 5 cases of stable disease (SD), and 5 cases of progressive disease (PD). The total effective rate (CR + PR) was 78.26%. The one year survival rate was 67.4%. For 3D-CRT group, there were 5 cases of complete response, 25 cases of partial response, 9 cases of stable disease, and 7 cases of progressive disease. The total effective rate was 65.2% and the one year survival rate was 52.2%. There was no significant difference in the total effective rate between C-RFA plus chemotherapy group and 3D-CRT plus chemotherapy group (P >0.05), but one year survival rate of C-RFA plus chemotherapy group was higher than that of 3D-CRT plus chemotherapy group (P <0.05). The difference between the activity of NKc and the ratio of T cell subgroups was significant before and after C-RFA therapy (P <0.05). Conclusions: The C-RFA combined with chemotherapy has a definite effects on advanced NSCLC. It might be used as one of the multidisciplinary therapies for NSCLC.

Intrasplenic heterotransplantation of in vitro cultured human fetal hepatic stem cells for treatment of acute liver injury in mice with severe combined immunodeficiency / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the in vivo colonization, migration, and differentiation of in vitro cultured human fetal hepatic stem cells (HSCs) following intrasplenic transplantation for treatment of acute liver injury in mice with severe combined immunodeficiency (SCID).</p><p><b>METHODS</b>Human fetal HSCs were isolated from the normal fetal liver (16-24 weeks) and purified, and the morphology of HSCs was observed under optical and transmission electron microscopes. The expressions of stem cell markers were examined in these HSCs by means of immunocytochemistry and flow cytometry. The passaged human fetal HSC suspension (0.2 ml) were injected into the spleen of SCID mice with acute liver injury induced by two-third partial hepatectomy, and 15, 30, 60, and 90 days after cell transplantation, immunohistochemistry was performed to examine the location and expressions of human hepatocytes, alpha1-AT and AFP antigen in the spleen and liver of the recipient SCID mice. PAS staining was used to examine the expression of glycogen and RT-PCR employed for detection of the expressions of AFP and albumin mRNA in the spleen of the mice on the scheduled time points.</p><p><b>RESULTS</b>Under optical microscope and transmission electron microscope, most of the HSCs were small, about 1/6 to 1/3 of the size of the hepatocyte, with relatively large nucleus-cytoplasm ratio and only small quantities of endocytoplasmic reticulum, chondriosome, and ribosome. Immunohistochemistry and flow cytometry identified positive expressions of AFP, Thy-1, C-kit, CD34 and CK19 in the HSCs, and after cell transplantation, positive expressions of human hepatocyte, alpha1-AT, and AFP antigen occurred in the liver and spleen of the recipient SCID mice. PAS staining confirmed the presence of glycogenosome in the spleen of the mice following cell transplantation. RT-PCR on days 30, 60, and 90 showed positive expressions of human AFP and albumin mRNA in the spleen of the mice.</p><p><b>CONCLUSION</b>Human fetal HSCs can survive and settle in the spleen and liver, and migrate to the damaged liver of the recipient mice after intrasplenic transplantation, with the capacity of proliferation and differentiation into hepatocytes in the recipient target organs.</p>

Screening and preliminary analysis of colorectal carcinoma-associated antigen genes / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen and identify the genes coding for colorectal carcinoma-associated antigen and analyze the bioinformation of their cDNA sequences.</p><p><b>METHODS</b>Immunoscreening of the cDNA phage-display library derived from human colorectal carcinoma was performed with autologous or allogeneic serum antibody from patients with colorectal cancer through SEREX approach. After amplification of the positive phage clones, the phage DNA was extracted and purified with Qiagen kit, and the fragment sizes of the cDNA of positive clones were identified by PCR and EcoR I and Hind III restriction endonucleases. The cDNAs of the positive clones were ligated into pUCm-T vector and sequenced. The bioinformation of cDNA sequences were analyzed against GenBank+EMBL+DDBJ+PDB Sequences Database.</p><p><b>RESULTS AND CONCLUSION</b>Eleven positive clones were obtained after immunoscreening, and the sizes of the cDNA fragments were 1100, 1300, 1000, 2000, 1200, 1200, 700, 900, 600, 1200 and 1000 bp, respectively, representing 9 antigen genes, including 7 with homology with the known genes. Among the 11 obtained positive clones, 3 were the same cDNA having homology with interferon-induced transmembrance protein-1 and possessing anti-proliferation effect; another 6 represented different genes, namely human BAC clone RP11-453E17 whose function have not been cleared, human cartilage-hair hypoplasia region gene responsible for cartilage-hair hypoplasia, human chromosome 5 clone CTD-2030B15 with insertion mutation, human gene similar to anti tumor necrosis factor-alpha antibody light-chain Fab fragment associated with tumor growth, mRNA of human beta-2-microglobulin in relation to tumor cell proliferation, and human aldolase A gene promoting tumor cell proliferation. The other two cDNA sequences were not identified for homology with currently known genes in GenBank, and their functions awaits further investigation.</p>