Overexpression of response gene to complement-32 promotes cytoskeleton reorganization in SW480 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct the recombinant plasmid pcDNA3.0-RGC32 and evaluate the effect of the response gene to complement-32 (RGC32) on cell cytoskeleton in vitro.</p><p><b>METHODS</b>The full-length cDNA of RGC32 was obtained by RT-PCR and inserted into the eukaryotic expression vector pcDNA3.0 to generate the recombinant plasmid pcDNA3.0-RGC32. After transfection of the recombinant plasmid into SW480 cells, the expression of RGC32 in the cells was detected by Western blotting. The cytoskeleton of SW480 cells was visualized before and after the transfection, and the changes in the cell migration ability was assessed by wound-healing assay.</p><p><b>RESULTS</b>The recombinant plasmid pcDNA3.0-RGC32 was successfully constructed. The expression of RGC32 was significantly increased in SW480 cells after transfection with pcDNA3.0-RGC32. Before the transfection, the microfilaments of SW480 cells were few and short without obvious polarity, but after the transfection, the microfilaments were increased and elongated with also an obvious polarity, and the invasive structures of lamellae and lamellipodia occurred. The migration ability of the cells was enhanced after transfection with pcDNA3.0-RGC32.</p><p><b>CONCLUSION</b>Overexpression of RGC32 can cause the reorganization of cytoskeleton and promotes the cell migration, which can be an important mechanism of RGC32 in promoting cancer metastasis.</p>

Toxicity of matrine in Kunming mice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the acute toxicity and assess the median lethal dose (LD50) of matrine in Kunming mice.</p><p><b>METHODS</b>Matrine at different doses were administered in Kunming mice via intraperitoneal injection, and the toxic reactions and LD50 of matrine was observed and determined.</p><p><b>RESULTS</b>The acute toxicity test of matrine indicated that the tolerable dose of matrine was above 80 mg/kg in Kunming mice, and the LD50 was 157.13 mg/kg (95%CI, 88.08-280.31 mg/kg). Morphological observation revealed degenerative changes of the nerve cells in the brain tissue of the mice.</p><p><b>CONCLUSION</b>The nervous system is the main target organ by the toxicity of matrine.</p>

Celecoxib enhances the lethal effects of bleomycin in human tongue squamous carcinoma cell line Tca8113 / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of COX-2 inhibitor celecoxib in enhancing the lethal effects of bleomycin in Tca8113 cell line.</p><p><b>METHODS</b>Tca8113 cells were treated with different concentrations of celecoxib and bleomycin for 24, 48, 72 h. Methyl thiazolyl tetrazolium assay was used to calculate cell growth inhibition rate and Jin Zheng Jun's method was used to evaluate the interaction of celecoxib and bleomycin on Tca8113 cells. Flow cytometry was used to evaluate the effects of combined use of celecoxib and bleomycin on cell cycle progress and apoptosis.</p><p><b>RESULTS</b>Low dose of celecoxib (10 micromol/L, < IC(50)) combined with bleomycin showed synergism or additive lethal effect on Tca8113 cell line. Celecoxib could notably enhance the inhibitory effect of bleomycin on Tca8113 cells by blocking cell cycle progress and thus resulting in the increasing G(0)/G(1) cells [(60.93 +/- 0.32)%] distribution and inducing apoptosis [(1.87 +/- 0.11)%].</p><p><b>CONCLUSIONS</b>Low doses of celecoxib could significantly enhance the lethal effect of bleomycin on Tca8113 cells by inhibiting cell growth and proliferation through blocking cell cycle progress and inducing apoptosis. The ways of these interactions on inhibiting Tca8113 cell growth were synergistic or/and additive.</p>

Expression of HSP27 in colorectal carcinoma and its relationship with lymphatic metastasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relationship between heat shock protein 27 (HSP27) expression and lymphatic metastasis of colorectal carcinoma (CRC).</p><p><b>METHODS</b>Immunohistochemistry was used to detect HSP27 expression in 68 specimens of human CRC. The expression of HSP27 mRNA and protein was also detected in two colorectal carcinoma cell lines with different lymphatic metastasis potentials by RT-PCR, Western blotting and immunohistochemistry.</p><p><b>RESULTS</b>HSP27 expression was not related to such clinicopathological factors as the patient's age, gender, histological grade, lymphatic metastasis or clinical stages (P>0.05), but overexpression of HSP27 showed significant inverse correlation to lymphatic metastasis of CRC (P=0.035). HSP27 mRNA and protein were expressed at low levels in SW620 cells with high potentials for lymphatic metastasis, and at high levels in SW480 cells that had low lymphatic metastasis potentials.</p><p><b>CONCLUSION</b>HSP27 expression can be inversely correlated to metastatic behavior of CRC cells, and may play a role in suppressing the metastasis of CRC.</p>

Expression of COX-2 and VEGF-C in squamous cell carcinoma of the tongue and its correlation to lymph node metastasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the coexpression of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor-C (VEGF-C) in human squamous cell carcinoma of the tongue (SCCT) and assess their correlations to neoangiogenesis and lymph node metastasis of the tumor.</p><p><b>METHODS</b>Tissue samples of primary SCCT and the metastatic lymph nodes were obtained from 46 patients undergoing surgical resections of SCCT for immunohistochemical detection of COX-2 and VEGF-C expressions.</p><p><b>RESULTS</b>The over-expression rates of COX-2 and VEGF-C was 82.61% and 73.91% in the primary tumor, respectively, significantly higher than those in normal oral mucosa. A significant correlation was found between the expression scores of COX-2 and VEGF-C with a concordance rate of 82.61% (P<0.01) and a kappa value of 0.495 according to Cohen's kappa test (P<0.01). High expression of VEGF-C, but not COX-2, was correlated to the presence of lymph node metastasis (P<0.05). The mean microvessel density was significant higher in tumors with strong positivity for COX-2 or VEGF-C than in tumors with only weak or negative expressions (P<0.001).</p><p><b>CONCLUSION</b>The expressions of COX-2 and VEGF-C are closely correlated in SCCT, and may have clinical value for assessing the prognosis, metastasis and invasion of SCCT, but the mechanism of these proteins in carcinogenesis in SCCT needs further investigation.</p>

Matrine-induced apoptosis in human colon adenocarcinoma SW620 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of matrine on the cell cycle and apoptosis in human colon adenocarcinoma SW620 cells and explore the possible mechanisms.</p><p><b>METHODS</b>The effect of matrine on cell proliferation was assessed using MTT assay, and the cell cycle arrest induced by matrine was determined by flow cytometry. The changes of cell morphology were observed through optical microscope, fluorescence microscope and electron microscope, and the cell apoptosis was detected using Annexin V-FITC apoptosis assay.</p><p><b>RESULTS</b>Matrine inhibited the proliferation of SW620 cells in a dose- and time-dependent manner. Compared with the control group, the matrine-treated cells showed increased cell percentage arrested in G 0/G1 phase with decreased S-phase cells. Morphologically, the SW620 cells treated with matrine exhibited cell shrinkage, cell size reduction, plasma condensation, cytoplasmic vacuolar changes, and formation of apoptotic body with also the presence of the signet-ring cells, all typical of apoptotic cells.</p><p><b>CONCLUSION</b>Matrine exposure of SW620 cells inhibits the cell proliferation, causes cell cycle arrest at G 0/G1 phase, and induces apoptosis in a dose- and time- dependent manner.</p>

Monitoring changes of serum protein markers in metastatic colorectal carcinoma model / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the changes of several protein markers in a metastatic colorectal carcinoma model by serum proteomic analysis.</p><p><b>METHODS</b>The pEGFP-N1 plasmid with enhanced expression of green fluorescence protein (EGFP) was transfected into human colon carcinoma cell line SW480 to obtain a stable SW480-EGFP cell line, the SW480-EGFP cells were then injected subcutaneously into nude mice. The harvested tumor cells were implanted orthotopically into the colon of the nude mice. Real-time tumor growth and metastasis formation were visualized by whole-body fluorescent imaging system. Serum samples at different metastatic stages were collected and differential proteomic profiles were investigated by two-dimensional gel electrophoresis (2DE) and matrix-assisted laser absorption/ionization time of flight mass spectrometry (MALDI-TOF MS).</p><p><b>RESULTS</b>The SW480- EGFP cells enabled to express EGFP stably. The rates of subcutaneous and orthotropic tumor formation were 100%. The metastasis rates to local lymph nodes, liver and lung were 100%, 40% and 30%, respectively. Furthermore, 5 differentially expressed proteins were analyzed by serum proteome technologies, including haptoglobin alpha chain, apolipoprotein E, apolipoprotein A-IV, Ig kappa chain V region chain L and transferrin.</p><p><b>CONCLUSIONS</b>Visualized metastatic model of colorectal carcinoma was successfully established. Several differentially expressed serum proteins collected at different stages after the occurrence of metastasis were identified. These differentially expressed proteins may be candidate serum biomarkers for diagnosis and therapeutic evaluation of colorectal carcinoma metastasis.</p>

Expression and diagnostic application of C4.4A protein in squamous cell carcinoma and adenocarcinoma / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the diagnostic utility of C4.4A gene expression in discriminating a squamous cell carcinoma (SCC) from an adenocarcinoma by immunohistochemistry.</p><p><b>METHODS</b>Immunohistochemical staining was performed to detect the expression of C4.4A protein in 157 cases of SCC and 177 cases of adenocarcinoma of various organs.</p><p><b>RESULTS</b>Overall, 141 of 157 cases of SCC strongly expressed C4.4A protein. In contrast, only 8 of 177 adenocarcinomas showed partial or scattered cell expression of C4.4A protein. The statistic difference between the two groups was highly significant (chi(2) = 244.93, P = 0.000), and also when the tumors were stratified according to the degree of differentiation (P = 0.000).</p><p><b>CONCLUSION</b>C4.4A protein expression may serve as a valuable tumor marker in discriminating a squamous cell carcinoma from an adenocarcinoma, and therefore, may greatly facilitate the differential diagnosis of an epithelial malignancy.</p>

Association of abnormal cyclooxygenase-2 gene expression with colorectal carcinoma metastasis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate cyclooxygenase-2 (COX-2) expression of in colorectal carcinoma cell lines and tissues and its clinical implications.</p><p><b>METHODS</b>SP immunohistochemistry was used to detect COX-2 protein in SW480 and SW620 cell lines and 50 primary colorectal carcinoma and 50 lymphoma metastasis carcinoma specimens. Real-time PCR was used to detect COX-2 mRNA expression in SW480 and SW620 cell lines.</p><p><b>RESULTS</b>SW480 and SW620 cell lines both highly expressed COX-2. The positive expression levels of COX-2 increased significantly in lymph node metastatic carcinoma in comparison with primary colorectal carcinoma (P<0.05). The expression COX-2 mRNA in SW620 cell line was higher than that of SW480 cell line, showing a mean expression increment of 2.268 folds in SW620 cell line relative to SW480.</p><p><b>CONCLUSION</b>Elevated COX-2 expression may be associated with lymph node metastases of colorectal carcinoma.</p>

Changes of biological characteristics of colon carcinoma cell line - LoVo induced by Kai1/CD82 transfection / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the influence of Kai1/CD82 transfection on the growth, adherence, separation and invasion potential of LoVo colon carcinoma cell line.</p><p><b>METHODS</b>Kai1/CD82 cDNA was transfected into LoVo cells, and a stable expressing clone was established. In vitro methodology was used to obtain the growth curve and also to detect the adherence, separation and invasion potential of the transfected LoVo cells, in comparison with those of control cells without transfection.</p><p><b>RESULTS</b>Compared with the control, no change was observed in the growth pattern of transfected LoVo cells. The numbers of adherent cells in the two groups were 0.08, 0.63, 0.83, 0.91 (x 10(5)) for the transfected cells and 0.04, 0.48, 0.71, 0.82 (x 10(5)) for the control cells respectively after 10, 20, 30, 40 minutes culture with shaking. The difference at 20, 30 and 40 minutes was statistically significant (P < 0.05). The separation rates of each group were 13%, 20%, 53% for the transfected cells and 11%, 28%, 60% for the control cells, respectively after 5, 10, 15 minutes culture with shaking. The difference at 10 and 15 minutes was statistically significant (P < 0.05). The aggregation rate of the transfected cells was higher than that of the control cells after culture with mild shaking for 5 hours (64.8% vs. 58.6%, P < 0.05). After co-incubation with endothelium cells ECV304, the number of invading cells decreased more in the transfected cells than that in the control cells (6.33/field and 17.67/field, P < 0.05).</p><p><b>CONCLUSION</b>Transfection expression of Kail/CD82 into LoVo cell line results in an increase of cell adherence and aggregation, but a diminished capability of separation and invasion, suggesting that the expression of Kai1/CD82 gene may inhibit the metastatic potential of colon carcinoma.</p>

Study on etiology and pathology of severe acute respiratory syndrome / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic characteristics of severe acute respiratory syndrome (SARS).</p><p><b>METHODS</b>Three autopsy cases were studied retrospectively. Routine HE stain was used to study all the cases. Part of the lung tissue specimens were studied further with Macchiavello's stain, viral inclusion body stain, reticulin and PAS stains, immunohistochemistry, thin sections with staining, light microscopy and transmission electronic microscope investigation.</p><p><b>RESULTS</b>The earliest symptom of all 3 cases was hyperpyrexia and followed by progressive dyspnea and appearance of lung field shadows in X rays findings. Pulmonary lesions included: bilateral and extensive consolidation, localized hemorrhage and necrosis, desquamative alveolitis and bronchitis, alveolar proliferation and desquamation, accumulation of protein exudates, mononuclear cells, lymphocytes, and plasma cells as well as hyaline membrane formation in alveoli and viral inclusion bodies were seen in the alveolus epithelial cells. The exudated organization tended to become glomeruloid organizing pneumonitis in a few avaoli. Lesions of the immune organs included: large patchy necrosis in the spleens and localized necrosis in the lymph nodes were seen. Bone marrow became restrained. There were lesions of systemic small vasculitis including edema of the perivascular tissue and vascular wall of the small veins with localized fibrinoid necrosis distributing in the heart, lungs, kidneys, adrenal glands and the striated muscles accompanying with mononuclear cells and lymphocytes infiltration. Thrombosis was seen in part of the small veins. In addition, there were also the systemic poisonous changes including: degeneration and necrosis of the parenchyma cells in lungs, liver, kidneys, heart and adrenals. Electronic microscopy demonstrated clusters of virus particles seen in the lung tissue.</p><p><b>CONCLUSION</b>SARS is a systemic disease. Lungs, immune system and systemic small vessels are the main target organs attacked by the virus. Extensive consolidation of lungs, formation of hyaline membrane to a large extent, respiratory distress and decrease of immune function are the main causes of death.</p>