For an advanced elucidation of mechanisms of nm23-H1 suppressive effects on metastasis of primary hepatocellular carcinoma (HCC), it is necessary to investigate the correlation between nm23-H1 expression and relative factors involved in the HCC invasion. In present report, full-length cDNA of nm23-H1 was subcloned into pBKCMV vector and transfected into HCC cell line to observe its effects on invasion, cytosolic free Ca2+ and Nras mRNA expression. The results showed that lower expression of N-ras and higher cytosolic free Ca2+ in transfected cell line were detected, while the potential of invasion was depressed. It suggests that the suppressive effects on HCC metastasis might interact with intracellular signal transduction which is essential for stimulating cell invasion.
Objective To examine the effects of newly designed LY52 on the expression of matrix metalloproteinases and invasive ability of hepatocellular carcinoma HepG2 cells. Methods The effects of LY52 on the proliferations of HepG2 cells were detected by MTT assay. Gelatin zymography and Western blot were used to detect the effects of LY52 on matrix metalloproteinase-2 expression in the cell line. Transwell chamber assay was used to detect the effects of LY52 on the invasion of the cells. Results No obvious inhibitory or cytotoxicity effects of LY52 was found in lower concentrations (lt;200 μg/ml) of LY52. Gelatin zymography and Western blot showed that matrix metalloproteinase-2 expression were inhibited by LY52 in a dose-dependent manner in HepG2 cells. Furthermore, transwell chamber assay showed that LY52 could significantly inhibit the invasion of the cell line in a dose-dependent manner.Conclusion The results suggest that LY52 may inhibit the invasion of hepatocellular carcinoma cells by suppressing the matrix metalloproteinase-2 activity.
【Abstract】Objective To study the effects of exogenous hyaluronidase on invasive and angiogenic potential of human breast cancer cell line ZR-75-30.MethodsThere were two groups in the study: the study group (hyaluronidase group) and the control group. The invasive potential and the angiogenic potential of human breast cancer cell ZR-75-30 were detected by the invasive model in vitro and technique of double-chamber co-culture that human breast cancer cell line ZR-75-30 and human umbilicus vein endothelium cell ECV-304 were co-cultured. ResultsThe penetrating number of tumor cell in the study group (70.625±11.64) was significantly higher than that in the control group (22.125±6.09),P＜0.01. The tube number from ECV-304 cell induced by ZR-75-30 cell in the study group (34.5±2.4) was significantly higher than that in the control group (8.5±1.5)， P＜0.01. ConclusionExogenous hyaluronidase can reinforth the invasive and angiogenic ability of breast cancer cells.
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.
【Abstract】ObjectiveTo study the effect of down-regulation of E-cadherin on the invasion ability of tumor cells. MethodsHuman pancreatic carcinoma cell line JHP-1 was treated with E-cadherin antisense oligodeoxynucleotied (ASODN). The immunocytochemistry, Western blot were used to detect the expression and the contents of E-cadherin in the tumor cells, and the invasive ability of tumor cells were evaluated by invasive-MTT assay. Results Treated with E-cadherin ASODN, the expression of E-cadherin on JHP-1 cells were reduced, and the protein contents were decreased as well compared with control groups and ODN group. The invasive ability of JHP-1 cells to the basement membrane was increased (P＜0.001) compared with ODN group and control group. ConclusionE-cadherin was related to the invasive ability of tumor cells.
Objective To detect the expression of KiSS-1 protein in papillary thyroid carcinoma, and to analyze its significance. Methods Paraffin-embedded specimens of 32 patients with thyroid papillary carcinoma and its adjacent cancer tissues were included in this study. Then the expression of KiSS-1 protein was detected by munohistochemistry and its relationship with clinical pathological features was analyzed. Results KiSS-1 protein mainly expressed in the cell membrane and cytoplasm. The expression of KiSS-1 protein was positive in adjacent tissues, but decreased or absent in cancer tissues in 32 patients. In the latter, there were 11 cases with positive expression (34.4%) and 21 cases with negative expression (65.6%), and the difference was statistically significant (χ2=31.256, Plt;0.001). The average value of KiSS-1 protein expression represented by absorbance (A) value (119.595 2) in cancer tissues was higher than that in adjacent tissues (174.805 0), t=34.429, Plt;0.001. The expression of KiSS-1 protein in cancer tissues was not related to patient gender (P=0.618) and age (P=0.061), but except TNM staging (P=0.034). The expression rate of KiSS-1 protein in cancer tissues with lymph node metastasis (4/4, 100%) was significantly higher than that without lymph node metastasis (7/28, 25.0%)， P=0.003. Conclusion The expression of KiSS-1 protein is decreased or absent in papillary thyroid carcinoma, which may be involved in tumorigenesis, invasion, and metastasis.
Objective To investigate the expression of c-met in tall cell variant of papillary thyroid carcinoma, and to compare it with other types of thyroid carcinoma and benign thyroid tissue. Methods The expressions of c-met in 60 cases of thyroid specimens were tested by immunohistochemical staining. Results The levels of expressed c-met in tall cell variant specimens were significantly higher than those in other types of papillary thyroid carcinoma and benign thyroid tissue. c-met expressions were significantly different in the following pairs of types: tall cell variant vs common papillary carcinoma of thyroid (P=0.000 1), tall cell variant vs follicular variant papillary thyroid carcinoma (P=0.000 1), and tall cell variant vs benign thyroid tissue (P=0.000 1). In addition, for all types of papillary carcinomas evaluated, c-met expression was significantly higher in specimens with extracapsular spread (P=0.010 0) and skeletal muscle invasion (P=0.020 0). Conclusion The high expression of c-met is a significant marker for tall cell variant papillary carcinoma of thyroid and its invasive behavior. This finding may explain the unusually aggressive behavior of this tumor and suggest a role for c-met in the early identification of patients with tall cell variant thyroid carcinoma.
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.
ObjectiveTo investigate the influence of EZH2 gene down-regulation by RNA interference on the proliferation and invasion of human glioma cell line U251. MethodsThe recombinant plasmid of small hairpin RNA targeting EZH2 gene was constructed, and transfected into gioma U251 cells by electroporation. The expression of EZH2 mRNA and protein in the cells was detected by using reverse transcriptase-polymerase chain reaction and Western blot respectively; the viability of cells was determined by using methyl thiazol tetrazo1ium assay; and the invasiveness of U251 cells was tested by Transwell cabin. ResultsThe expression levels of EZH2 mRNA in U251 cells were detected in a significantly lower proportion in the EZH2-shRNA group (0.19±0.02) than that in the untransfected group (1.13±0.05) and the control-shRNA-GFP group (1.15±0.05). The expression levels of EZH2 protein in U251 cells were detected in a significantly lower proportion in the EZH2-shRNA group (0.20±0.02) than that in the untransfected group (1.03±0.03) and the control-shRNA-GFP group (0.97±0.06). The proliferation rates in EZH2-shRNA group were significantly decreased as compared with those in the untransfected group and control-shRNA-GFP group (24 hours after transfection:60.13%±3.15%, 100.00%±9.31%, 100.03%±9.35%; 48 hours after transfection:53.01%±3.14%, 100.00%±9.13%, 99.58%±9.27%; P<0.05) and Transwell cabin suggested that the invasiveness of U251 cells was significantly decreased (46.00±2.82, 60.67±5.71, 61.00±2.48; P<0.01). ConclusionEZH2-targeted RNA interference can reduce the proliferation and invasion of human glioma cells, which suggests that EZH2 shRNA may be a potential gene therapeutic target of human glioma.