miRNA-125b over-expression for promoting proliferation and invasion ability of lung cancer A549 cells and its mechanism / 重庆医学

Objective To study the influence of miRNA-125b over-expression on proliferation and invasion ability of lung cancer A549 cells and its mechanism.Methods A549 cells were divided into 3 groups:miRNA-125b group(transfected with miRNA-125b mimics),NC group(transfected with NC mimics) and blank group(same volume of GIBCO serum mixed with transfection agent).The transfection and expression efficiency of miRNA-125b was detected with Q-PCR,the proliferation ability was detected with MTT,and the invasion ability was detected with the transwell chamber test.The expression level of BMF in A549 cells was detected with Western blot.Results Compared with the blank group,the expression level of miRNA-125b,proliferation ability and invasion ability in the miRNA-125b group were increased(P＜0.05);while the above indexes in the NC group demonstrated no significant change(P＞0.05).Compared with the blank group,the expression level of BMF in the miRNA-125b group was decreased (P＜0.05);while which in the NC group had no significant change(P＞0.05).Conclusion miRNA-125b can promote the proliferation and invasion ability of A549 cells via inhibiting the expression of BMF.

Interferon Regulatory Factor 5 Inhibits Invasion Ability of Nasopharyngeal Carcinoma by Reducing PARP-1 / 中山大学学报(医学科学版)

[Objective]To investigate whether IRF5 can inhibit invasion ability of nasopharyngeal carcinoma by re-ducing PARP-1(poly(ADP-ribose)polymerase-1).[Methods]Forty-six specimens of nasopharyngeal carcinoma and 51 specimens of normal tissue were confirmed by pathologically in this study.The expression of IRF5 and PARP-1 in naso-pharyngeal carcinoma tissues and normal tissues was detected by immunohistochemistry.The IFR5 overexpression plasmid was transfected into the nasopharyngeal carcinoma cell line CNE-2,quantitative PCR and immunoblotting was used to value the expression of IRF5 after transfection.The wound healing and transwell assay was used to investigate the invasion ability. The expression of PARP-1 was valued by quantitative PCR and immunoblotting after over-expression of PFR5.[Results]The results showed that the expression of IRF5 in cancer tissues was lower than that in normal tissues,but the PARP-1 expression was opposite. The IRF5 overexpressing cell line CNE-2/IFR5 was established. The healing rate of CNE-2/IFR5 cells was lower than that of the control cells(P<0.01). Transwell experiments revealed that the number of CNE-2/IFR5 cells passing through the basement membrane was smaller than that of the control group(P<0.01),suggest-ing that up-regulation of IFR5 could inhibit the invasiveness of nasopharyngeal carcinoma cells.Over-expression of IFR5 led to reduced PARP-1 mRNA and protein(P<0.01).Besides,elevation of PARP-1 can prevent IRF5-induced changes of invasion ability.[Conclusion]Therefore,we speculated that IRF5 can inhibit invasion ability of nasopharyngeal carci-noma by reducing the expression of PARP-1.This study provided a new target for inhibiting the invasion ability of naso-pharyngeal carcinoma based on IRF5.

VCAM-1 improves migration and invasion of human glioma cell lines / 基础医学与临床

Objective To investigate the effects of VCAM-1 on migration and invasion of glioma cell lines . Methods The techniques of lentivirus pSGU6/GFP/Neo-based VCAM-1 shRNA and EF1 a-GFP/puro-based VCAM-1 expression vector, the scratch wound healing migration and transwell invasion assays , and the Western blot and cell staining were applied to observe the effects of VCAM-1 expression levels on migration and invasion of glioma cell line cells.There are four groups in T98G cells including control, vector, scramble and shRNA-VCAM-1 groups and three groups in U251 cells covering control, vector and VCAM-1 overexpressed groups ( n=6 per group) .Results The stabled glioma cell lines of T98 G cells with down-regulated VCAM-1 and U251 cells with VCAM-1 overexpression were established by using lentivirus-based VCAM-1 shRNA and expression vector.The ability of scratch wound healing (migration activity) decreased significantly (P<0.01) in T98G cells with lower VCAM-1 expression levels, while the migration activity was obviously improved in U251 cells with overexpressed VCAM-1 ( P <0.05 ) .Similarly, the invasion ability was significantly inhibited ( P <0.05) in T98G cells with silenced VCAM-1, as well as VCAM-1 overexpression could enhance the invasion ability of U251 cells ( P<0.01 ) .Conclusions VCAM-1 improves the migration activity and invasion ability of human glioma cell line cells.

Legionella dumoffii Tex-KL Mutated in an Operon Homologous to traC-traD is Defective in Epithelial Cell Invasion / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To understand the mechanism of invasion by Legionella dumoffii.</p><p><b>METHODS</b>The L. dumoffii strain Tex-KL was mutated using the Tn903 derivative, Tn903dIIlacZ. After screening 799 transposon insertion mutants, we isolated one defective mutant. We then constructed the gene-disrupted mutant, KL16, and studied its invasion of and intracellular growth in HeLa and A549 cells, and in A/J mice survival experiments. The structure of traC-traD operon was analyzed by RT-PCR.</p><p><b>RESULTS</b>The transposon insertion was in a gene homologous to Salmonella typhi traC, which is required for the assembly of F pilin into the mature F pilus structure and for conjugal DNA transmission. Results from RT-PCR suggested that the traC-traD region formed an operon. We found that when the traC gene was disrupted, invasion and intracellular growth of L. dumoffii Tex-KL were impaired in human epithelial cells. When mice were infected by intranasal inoculation with a traC deficient mutant, their survival significantly increased when compared to mice infected with the wild-type strain..</p><p><b>CONCLUSION</b>Our results indicated that the traC-traD operon is required for the invasion and intracellular growth abilities of L. dumoffii Tex-KL in epithelial cells.</p>

Changes of TIZ expression in epithelial ovarian cancer cells

Objective:To study the change ofTIZ expression in epithelial ovarian cancer cells.Methods:HO8910 cells were transinfected with siRNA to inhibit the expression ofTIZ. pcDNA3.1-TIZ vectors were combined to increase theTIZ expression level.The cell viability, colony forming efficiency and cycle distribution ofHO8910,HO8910/NC,HO8910/pcDNA3.1-NC,HO8910/TIZ-573 andHO8910/pcDNA3.1-TIZ were compared, and the invasion rate, migration rate and adhesion rate between5 groups of cells were compared.Results:Compared with those ofHO8910,HO8910/NC andHO8910/pcDNA3.1-NC, the cell viability, colony forming efficiency and cell cycle distribution ofHO8910/TIZ-573 were increased, while the indexes ofHO8910/pcDNA3.1-NC were decreased with statistical significant difference(P0.05). Conclusions:The expression ofTIZ can inhibit the proliferation of epithelial ovarian cancer cells.

Changes of TIZ expression in epithelial ovarian cancer cells

OBJECTIVE@#To study the change of TIZ expression in epithelial ovarian cancer cells.@*METHODS@#HO8910 cells were transinfected with siRNA to inhibit the expression of TIZ. pcDNA3.1-TIZ vectors were combined to increase the TIZ expression level. The cell viability, colony forming efficiency and cycle distribution of HO8910, HO8910/NC, HO8910/pcDNA3.1-NC, HO8910/TIZ-573 and H08910/pcDNA3.1-TIZ were compared, and the invasion rate, migration rate and adhesion rate between 5 groups of cells were compared.@*RESULTS@#Compared with those of HO8910, HO8910/NC and HO8910/pcDNA3.1-NC, the cell viability, colony forming efficiency and cell cycle distribution of HO8910/TIZ-573 were increased, while the indexes of H08910/pcDNA3.1-NC were decreased with statistical significant difference (P0.05).@*CONCLUSIONS@#The expression of TIZ can inhibit the proliferation of epithelial ovarian cancer cells.

Changes of TIZ expression in epithelial ovarian cancer cells

Objective: To study the change of TIZ expression in epithelial ovarian cancer cells. Methods: HO8910 cells were transinfected with siRNA to inhibit the expression of TIZ. pcDNA3.1-TIZ vectors were combined to increase the TIZ expression level. The cell viability, colony forming efficiency and cycle distribution of HO8910, HO8910/NC, HO8910/pcDNA3.1-NC, HO8910/TIZ-573 and H08910/pcDNA3.1-TIZ were compared, and the invasion rate, migration rate and adhesion rate between 5 groups of cells were compared. Results: Compared with those of HO8910, HO8910/NC and HO8910/pcDNA3.1-NC, the cell viability, colony forming efficiency and cell cycle distribution of HO8910/TIZ-573 were increased, while the indexes of H08910/pcDNA3.1-NC were decreased with statistical significant difference (. P0.05). Conclusions: The expression of TIZ can inhibit the proliferation of epithelial ovarian cancer cells.

Prevalence of virulence and cytolethal distending toxin (CDT) genes in thermophilic Campylobacter spp. from dogs and humans in Gyeongnam and Busan, Korea / 대한수의학회지

The prevalence of thermophilic Campylobacter (C.) spp. in stray, breeding, and household dogs was 25.2, 12.0, and 8.8%, respectively. C. jejuni and C. upsaliensis were the predominant Campylobacter spp. from household dogs. cdtA, cdtB, and cdtC were detected by PCR in all isolates. Despite the high cytolethal distending toxin (CDT) gene prevalence, only 26 (31%) C. jejuni strains and one (15.3%) C. coli strain showed evidence of CDT production in HEp-2 cell cytotoxicity assays. Virulence-associated genes detected in the C. jejuni and C. coli isolates were cadF, dnaJ, flaA, racR, ciaB, iamA, pldA, virB11, ceuE, and docC. cadF, dnaJ, flaA, and ceuE were found in all C. jejuni and C. coli isolates. When detecting Guillain-Barre syndrome-associated genes (galE, cgtB, and wlaN), galE was identified in all isolates. However, cgtB and wlaN were more prevalent in C. jejuni isolates from humans than those from dogs. Adherence and invasion abilities of the C. jejuni and C. coli strains were tested in INT-407 cells. A considerable correlation (adjusted R2 = 0.678) existed between adherence and invasion activities of the Campylobacter spp. isolates.

Dos aislamientos de Salmonella Infantis multirresistentes se comportan como hipoinvasivos pero con elevada proliferación intracelular

En este trabajo se investigó la presencia de determinantes característicos de plásmidos de virulencia en dos aislamientos clínicos de Salmonella Infantis portadores de plásmidos de multirresistencia. Además, se estudió la capacidad de invasión y proliferación en células eucariotas no fagocíticas. Ninguno de los aislamientos de S. Infantis mostró los determinantes genéticos que caracterizan a los plásmidos de virulencia para este género (operón spv). Los ensayos de invasión sobre líneas celulares eucariotas mostraron que los aislamientos de S. Infantis presentan una capacidad de invasión disminuida pero persisten y proliferan en el citoplasma, independientemente de utilizar una línea celular permisiva (HeLa) o no permisiva (NRK) para tal fin. Finalmente, no se observaron indicios microscópicos que podrían hacer sospechar un efecto bactericida de estas líneas celulares sobre los aislamientos estudiados.

Two multidrug-resistant Salmonella Infantis isolates behave like hypo-invasive strains but have high intracellular proliferation. In this work, plasmid-encoded virulence factors in two Salmonella Infantis isolates carrying multiresistance plasmids were investigated. In addition, their invasion and proliferative ability in non-phagocytic cells was studied. None of them showed the typical determinants of virulence plasmids (spv operon). The invasion assays of S. Infantis isolates on eukaryotic cells showed a decreased ability to Invade but they remained and proliferated In the cytoplasm regardless of having used a permissive (HeLa) or non-permissive (NRK) cell line. Finally, there was no microscopic evidence suggesting a bactericidal effect of these eukaryotic cell lines on the Isolates tested.

Invasion ability and collagenolytic activity of human glioma cells in vitro / 吉林大学学报(医学版)

Objective To investigate the invasion ability and collagenolytic activity of human glioma cells in vitro.Methods Boyden chamber invasion assay was employed to evaluate the cell migration ability of human malignant glioma cell line U87MG in vitro and the effect of conditioned medium of U87MG cells on matrix collagenolytic activity was tested by agar-gelatin gel.Results The number of U87MG glioma cells migrating through the Matrigel-coated membrane was more than that of addition of EDTA or anti-MMP-2 antibody in U87MG glioma cells(P0.05).EDTA or anti-MMP-2 antibody markedly inhibited the number of the migrated cells,the inhibitory rates were 79.2% and 77.1%,respectively. The conditioned medium collected from the U87MG cells showed an increase in the transparent ring area on agar-gelatin gel.Inhibition of MMP-2 enzymatic activity by EDTA or anti-MMP-2 antibody reduced the transparent ring area on agar-gelatin gel.Conclusion Both invasion ability and collagenolytic activity of U87MG cells are depended on MMP-2,suggesting that MMP-2 plays an important role in glioma invasiveness.

Inhibitory effect of VEGFR-3-targeted siRNA against adherence and invasion of human colon cancer cells / 中国肿瘤生物治疗杂志

Objective:To construct a small interfering RNA(siRNA)expression vector(psiRNA-VEGFR-3)targeting vascular endothelial growth factor receptor 3(VEGFR-3)and to investigate the effects of VEGFR-3 siRNA on the adherence and invasion of human colon cancer cells.Methods:A siRNA expression vector(psiRNA-VEGFR-3)targeting VEGFR-3 were constructed and was used to transfect LoVo cells via lipofectamine 2000.The mRNA and protein expression of VEGFR-3 were examined after transfection by reverse transcriptase polymerase chain reaction(RT-PCR)and Western blotting,respectively.The tumor adhesion ability was detected by cell-matrix adhesion experiment and the invasion ability of tumor cells was evaluated by millicell chamber model.Results:The VEGFR-3 siRNA expression vector was successfully constructed.The expression of VEGFR-3 mRNA and protein was inhibited after psiRNA-VEGFR-3 transfection.Seventy-two hours after psiRNA-VEGFR-3 transfection,Western blotting assay showed that the expression of VEGFR-3 protein was decreased from(1.26?0.19)to(0.39 s0.12)(P

Effects of antidiabetic drug metformin on the migration and invasion abilities of human lung adenocarcinoma A549 cell line in vitro / 中国癌症杂志

Background and purpose:There is growing evidence that metformin,a commonly used drug in the treatment of type Ⅱ diabetes,may impede the growth of human tumors.However in a recent study it was found that metformin treatment might result in promotion of the angiogenic phenotype and increase tumorigenic progression.In this study we investigated the effects of metformin on the migration and invasion abilities of human lung adenocarcinoma cell line A549 in vitro and explored the possible underlying mechanisms.Methods:A549 cells were treated with 0.5,2 and 8 mmol/L metformin for 72 hrs.The laterad-migration and invasion abilities of the cells in vitro were measured by scratch assay and Boyden-Chamber assay,respectively.Expressions of MMP2 and MMP9 mRNA of the cells before and after metformin treatment were measured by Real-time PCR.Results:The migration rate of A549 cells was increased after treated by metformin at the concentration of 8 mmol/L.The invasion ability was also signifi cantly increased by 8 mmol/L metformin treatment from 37.4?4.6 to 59.8?7.2(P