ObjectiveTo investigate the expression of β-catenin and Galectin-3 protein in human cervical carcinoma and its clinical pathological significance.
MethodsEighty-three cervical specimens were collected from January 2010 to June 2013. By immunohistochemical method, β-catenin and Galectin-3 expression of the 83 cases of cervical carcinoma, 45 cases of intraepithelial neoplasia (CIN) and 25 normal cervix tissue (control) were detected.
ResultsThe positive expression rate in cervical carcinoma of β-catenin and Galectin-3 was respectively 74.70% and 81.93%, which was significantly higher than that in intraepithelial neoplasia (ⅠandⅡ) and normal cervical tissue (P<0.05). Compared with cervical cancer, the expression of β-catenin and Galectin-3 in CINⅢ had no statistical significance (P>0.05). The positive expression of β-catenin was significantly correlated with histological grade of cervical cancer tissue, International Federation of Gynecology and Obstetrics grade and lymph node metastasis (P<0.05). The positive expression of Galectin-3 was closely related to histological grade of cervical cancer tissue and lymph node metastasis (P>0.05). Both β-catenin and Galectin-3 expression had no relationship with other clinical pathological parameters, such as age of patients, tumor size and pathological pattern of tumor (P>0.05). β-catenin expression had significant positive correlation with that of Galectin-3 (r=0.327, P=0.002).
ConclusionThe expression of Galectin-3 and β-catenin increases obviously and is associated with abnormal clinical parameters (invasion or metastasis) in patients with cervical cancer. Galectin-3 and β-catenin may act as cancer metastasis and prognostic indicators in these patients.
Objective To summarize the clinical features, predisposing factors, diagnosis, therapeutic outcome, and prognosis of invasive pulmonary fungal infection( IPFI) . Methods 90 cases with pathologically proved IPFI, admitted in non-intensive care unit in Xiangya Hospital from January 2005 to February 2012, were retrospectively analyzed. Results The pathogenic examination revealed Aspergillosis in 56 cases( 62. 2% ) , Cryptococcus in 18 cases( 20. 0% ) , Mucormycosis in 6 cases( 6. 7% ) , and Histoplasma in 6 cases( 6. 7% ) , etc. The underlying diseases were reported in 87 cases, and mainly included COPD, pulmonary tuberculosis, and diabetes mellitus. Cough and expectoration were the common clinical symptoms. 49 patients ( 54. 4% ) received long-term and broad-spectrum antibiotic therapy. The CT results revealed masses type in 25 cases( 27. 8%) , nodule lesions type in 15 cases( 16. 7% ) , lung consolidation type in 22 cases( 24. 4% ) , cavity type in 22 cases( 24. 4% ) , aspergilloma type in 6 cases( 6. 7% ) . 47 patients were clinical diagnosed with IPFI before biopsy with preliminary diagnosis accordance rate of 52. 2% . 31 cases ( 34. 4% ) underwent surgical resection of pulmonary lesions, and no recurrence was detected over two-year follow up. 56 cases ( 62. 2% ) received systemic anti-fugal therapy, and 43 cases( 76. 8% ) were cured or significantly improved. 3 cases ( 3. 3% ) refused any therapy. Conclusions The most frequently isolated pathogen of IPFI is Aspergillosis. The mainly underlying diseases are COPD, pulmonary tuberculosis, and diabetes mellitus. Long-termand broad-spectrum antibiotic therapy may be the major risk factor. Pathological examination is needed for final diagnosis. Surgical procedure can achieve optimal prognosis.
ObjectiveTo explore the effects of silencing the expression of ubiquitin-conjugating enzyme E2T (UBE2T) gene on proliferation, apoptosis, migration and invasion abilities of A549 cells.MethodsA549 cells were cultured in vitro. Three sets of shRNA-UBE2T plasmid vectors (UBE2T-shRNA1 group, UBE2T-shRNA2 group, UBE2T-shRNA3 group) and shRNA-NC (shRNA-NC group) were constructed, respectively. A549 cells were transfected with lipofection transfection. The cells transfected with empty vector were enrolled as control (control group). The transfection efficiency was detected by RT-PCR. The effects of silencing the expression of UBE2T gene on biological behaviors (proliferation, apoptosis, migration, and invasion) of lung cancer A549 cells were detected by clone formation assay, flow cytometry, Transwell assay and scratch test. The expression of proliferation and apoptosis related proteins, and expression of PI3K/AKT signaling pathway proteins were detected by Western blot. ResultsAfter transfection, expression level of UBE2T mRNA in UBE2T-shRNA1 group, UBE2T-shRNA2 group and UBE2T-shRNA3 group was significantly down-regulated (all P<0.05), whose down-regulation was the most significant in UBE2T-shRNA3 group (P<0.05). Compared with control group and shRNA-NC group, clone formation rate, number of invasion A549 cells, scratch healing rate, Ki67 expression, PCNA expression, p-PI3K/PI3K ratio and p-AKT/AKT ratio were significantly decreased in UBE2T-shRNA3 group (P<0.05), while A549 apoptosis rate, Bax/Bcl-2 ratio and cleaved caspase-3/caspase-3 ratio were significantly increased (P<0.05). There were no significant differences in the above indexes between control group and shRNA-NC group (P>0.05). ConclusionsThe shRNA interfering with UBE2T is reliable to construct the model of A549 cells with stable low-expression UBE2T. Down-regulation of UBE2T expression can promote apoptosis of A549 cells, inhibit their proliferation, invasion and migration abilities. The mechanism may be related to inhibiting the activation of PI3K/AKT signaling pathway.
Objective To observe the effect of epidermal growth factor (EGF) on the proliferation, adhesion, invasiveness and the activation of nuclear factor-κB (NF-κB), matrix metalloproteinases (MMPs) expression and explore related mechanisms in pancreatic cancer cells. Methods Cell invasion assay, proliferation assay and adhesion assay were used to examine the proliferation, adhesion and invasiveness of pancreatic cancer cells, respectively. NF-κB activity was detected by electrophoretic mobility shift assay (EMSA), and MMPs protein and mRNA expressions were investigated by gelatin zymography, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results EGF increased the invasiveness of pancreatic cancer cell in a dose-dependent manner (P＜0.05), but did not affect cell proliferation or adhesion. The expressions of MMP-9 mRNA and protein significantly increased after induction by EGF and were highest when EGF concentration was 50 ng/ml, while there was no effect on the expressions of MMP-2 mRNA and protein. Furthermore, NF-κB activity increased with increased concentration of EGF in a concentration-dependent manner (P＜0.05). In addition, NF-κB activity and the expressions of MMP-9 mRNA and protein by pretreatment with both pyrrolidine dithiocarbamate (PDTC) and EGF decreased when compared that by pretreatment with EGF alone. The invasiveness of pancreatic cancer cell by pretreatment with both PDTC and EGF decreased when compared that by pretreatment with EGF alone and nothing (P＜0.05).Conclusion The findings indicate that the NF-κB-mediated MMP-9 induction is essential for EGF-induced invasiveness in pancreatic cancer cells, which can be inhibited by PDTC.
ObjectiveTo explore the expression of collagen Ⅳ in breast cancer and its clinical significance. We analyzed the correlation of the results with other prognostic parameters which included tumor size, status of estrogen receptor, axillary nodal status, TNM grade, and 5 years survival. The expression of collagen Ⅳ in 93 cases of human primary breast cancer as well as 5 cases of benign breast masses were examined.MethodsUsing monoclonal antibodies of collagen Ⅳ, the expression of collagen Ⅳ in breast masses were detected with immunohistochemical technique (LSAB).ResultsThe absent expression of collagen Ⅳ in the tumor masses was correlated with axillary lymph node involvement, tumor size and poor prognosis (5 years survival). The patients who had no expression of collagen Ⅳ in tumor masses had a shorter survival. ConclusionThe expression of collagen Ⅳ in tumor samples are correlated with axillary node involvement and prognosis. Collagen Ⅳ would be helpful for evaluation of invasion and treatment in breast carcinoma.
ObjectiveTo study the expression of c-Met in colorectal carcinoma cells and the effect of hepatocyte growth factor (HGF) on proliferation and invasion of colon carcinoma cells SW480.
MethodsReal-time PCR and Western blot methods were respectively used to detect the expressions of c-Met mRNA and protein in the different colorectal carcinoma cells in order to screen the high c-Met expression cells. The SW480 cells were incubated with different concentrations (0, 20, 40, and 70 ng/mL) HGF. MTT assay and Transwell test were used to evaluate the effects of proliferation and invasion in the SW480 cells.
Results①The c-Met was expressed in each colorectal carcinomar cells, especially highly expressed in the colon carcinoma cells SW480 in vitro.②MTT assay showed that the HGF could promote the proliferation of SW480 cells in a dose-dependent manner with some extent.③Transwell test showed that the HGF could increase the invasion of SW480 cells.
ConclusionThe c-Met is highly expressed in colorectal carcinoma cells and HGF could promote proliferation and increase invasion of colorectal carcinoma cells in vitro.
ObjectiveTo explore the influence mechanism of proliferation and invasion in colon cancer cell after silence of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) gene.
MethodsRT-PCR or Western blot method was used to detect the expression of PTEN mRNA or protein among four colon cancer cell lines (HT-29, WiDr, CaCo-2, and Colo320 cell lines). small interfering RNA (siRNA) was used to synthetize PTEN siRNA and transfect it into colon cancer cells. The expression of PTEN protein after transfecting was detected by Western blot. WsT-1 and invasion assay were used to examine the effects of PTEN siRNA silence on proliferation and invasion in colon cancer cells.
ResultsPTEN mRNA and protein were expressed in all the four colon cancer cell lines. After PTEN siRNA transfected into the colon cancer cells, the expressions of PTEN proteins were inhibited in all the four colon cancer cell lines (P < 0.01), and the proliferation and invasion of colon cancer cells were enhanced significantly (P < 0.01).
ConclusionsPTEN siRNA play an important role in metastasis process of colon cancer via enhanced its proliferation and invasion. Therefore, the understanding biologic mechanisms for regulation of PTEN might enable better molecular target therapy of treating the colon cancer patients with metastasis.
ObjectiveTo investigate effect of small interfering RNA (siRNA) targeting inhibition of growth hormone receptor (GHR) on proliferation and invasion of human liver cancer line SMMC-7721.
MethodsSMMC-7721 cells were transfected with siRNA targeting human GHR by GenMuteTM transfection regent.The cells were divided into three groups:blank control group (non-transfected siRNA),negative control group (transfected with non-specific siRNA),and specificity transfected group (transfected with expression specifically interfere by GHR siRNA).the relative expression of GHR mRNA was detected by using real-time PCR.the expression of GHR protein was detected by using Western blot.The cell proliferation was assessed by CCK-8 assay.And the ability of invasion was examined by Transwell assay.
ResultsThe expressions of GHR mRNA and GHR protein in the specificity transfected group were significantly lower than those in the blank control group (P<0.05) and the negative control group (P<0.05).Compared with the blank control group and the negative control group,the absorbance value and the number of migrating cells of SMMC-7721 cells were decreased obviously (P<0.05) in the specificity transfected group.
ConclusionsiRNA targeting human GHR could reduce capability of proliferation migration and invasion of SMMC-7721 cells.
Objective To study the expressions of Wnt5a, MMP2, and MMP14 in the primary lesions of gastric cancer and the influences on clinicopathologic features. Methods The expressions of Wnt5a, MMP2, and MMP14 in the specimens of 106 patients with gastric cancer and 39 patients from the adjacent normal gastric tissues were detected by immunohistochemical staining, χ2 test and non-parametric test were used to analyze the relationships among them and between them and their influences on the clinicopathologic features. Results Extensive expressions of Wnt5a, MMP2, and MMP14 were demonstrated in the gastric cancer, which were significantly higher than those in the normal gastric tissues respectively (Plt;0.05). Positive expression of Wnt5a was associated with larger tumor diameter, deeper depth of invasion, higher degree of regional lymph node metastasis, later TNM stage, and higher rate of lymph node metastasis (Plt;0.05). In addition, Wnt5a expression was also associated with lymphatic infiltration and vascular infiltration (Plt;0.05). The expressions of MMP2 and MMP14 were associated with lymphatic infiltration, but not with vascular infiltration. Higher expressions of MMP2 and MMP14 were correlated with deeper tumor invasion, higher degree of regional lymph node metastasis, later TNM stage, and higher rate of lymph node metastasis (Plt;0.05). In addition, higher expression of MMP2 possesed greater tumor diameter (Plt;0.05). Spearman rank correlation analysis revealed the positive relation between Wnt5a and MMP2 （rs=0.240, P=0.014）, Wnt5a and MMP14 （rs=0.251, P=0.010）, as well as MMP2 and MMP14 （rs=0.444, P=0.000）. Conclusion Higher expressions of Wnt5a, MMP2, and MMP14 seem to promote invasion and metastasis of gastric cancer, and there are positive relations among their expressions.
ObjectiveTo investigate the effect and mechanism of silent information regulator 1 (SIRT1)in invasion of human gastric cancer.
MethodsThe expressions of SIRT1 protein and vascular endothelial growth factor A (VEGF-A) protein in 46 cases of gastric cancer were tested by immunohistochemical SP method; the effect of expressions of SRIT1 and VEGF-A protein on prognosis of gastric cancer was analyzed by Kaplan-Meier test; the expressions of SRIT1 and VEGF-A protein in human gastric mucosa GES-1 cells and SGC7901 cells were tested by Western blot method; after the interference of siRNA on SIRT1 gene, expressions of SRIT1 and VEGF-A protein were also tested by Western method, and the invasion ability was determined by Transwell test.
ResultsCompared with normal gastric mucosa tissues, expression levels of SIRT1 and VEGF-A protein of gastric tissues were both higher (P < 0.050), and survival situation of patients with SIRT1-positive (P=0.001) or SIRT1-positive and VEGF-A-positive (P=0.006) were both bad, but there was no significant difference on the relationship between prognosis of gastric cancer and expression of VEGF-A protein (P=0.091). Expression levels of SITR1 protein (P=0.010) and VEGF-A protein (P=0.020) in GES-1 cells were both higher than those of SGC7901 cells. In siRNA positive group, expressions of SIRT1 and VEGF-A protein (P=0.010) of SGC7901 cells down-regulated, and invasion ability decreased (P=0.000).
ConclusionsSIRT1 gene may promote the expression of VEGF-A protein and the invasion ability of gastric cancer, it may be a therapeutic target of invasion inhibition for gastric cancer.
