Effects of the F10 gene overexpression and silence on the expressions of invasion-related proteinases in choriocarcinoma cell line JAR / 医学研究生学报

Objective The F10 gene was found in the initial stage of our study to be highly expressed in hydatidiform mole. The aim of this study was to investigate the effects of theoverexpression or depletion of F10 on the invasiveness of the choriocarcinoma cell line JAR and explore the relationship of F10 expression with the invasiveness and metastasis ofchoriocarcinoma. Methods Using cell transfection and RNA interference technology, we established JAR choriocarcinoma cell lines with stablyoverexpressedor silenced F10 gene.We randomly and equally assigned 30 SPF nude mice into the three groups to receive the injection of JAR cellswith overex-pressed F10 ( F10 overexpression group) , untreated JAR cells ( control group) , and JAR cells with silenced F10( F10 silence group) . At 5 weeks after the JAR cell injection, we harvested the subcutaneous tumor tissues from the mice, determined the expressions of ma-trix metalloproteinases (MMP), tissue inhibitors of metalloproteinase-1(TIMP-1), and plasminogen activator inhibitor-1(PAI-1), and compared by Western blot and immunohistochemistry, and compared the expressions among the three groups of mice. Results Im-munohistochemistryshowed significantlyup-regulated expressions of MMP-2, -8, -11, -16, and-19 and down-regulated expressions of TIMP-1 and PAI-1 in the subcutaneous tumor tissues in the F10 overexpression group as compared with the control and F10 silence groups ( P<0.05) .Western blot also exhibitedextremely significantly up-regulated expressions of MMP-2,-8,-11,-16, and -19 pro-teins anddown-regulated expressions of TIMP-1 and PAI-1 proteins in the F10 overexpression groupin comparison with the control and F10 silence groups( P<0.01 ) . Conclusion By up-regulating the expressions of MMPs and down-regulating the expressions of TIMP-1 and PAI-1, the F10 gene might be an upstream stimulating factor involved in the proliferation, invasiveness, and metastasis of-choriocarcinoma cells.

Role of hydatidiform mole-related gene F10 in the tumorigenicity of choriocarcinoma cell line JEG-3 / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of the hydatidiform mole-related gene F10 in the tumorigenicity of choriocarcinoma cell lines JEG-3 in nude mice.</p><p><b>METHODS</b>Choriocarcinoma JEG-3 cell lines with stable F10 gene over-expression and F10 gene silencing were established using cell transfection and RNA interference techniques, respectively. Thirty SPF nude mice (4-5 weeks old) were equally randomized into F10 over-expression group, control group, and F10 gene-silenced group for subcutaneous injection of 0.2 ml cell suspension (5 × 10⁷ cells) of F10 gene over-expressing JEG-3 cells, non-treated JEG-3 cells, and F10 gene-silenced JEG-3 cells, respectively. The mice were observed and weighed every 3-4 days, and the tumor formation time was recorded to draw the tumor growth curve and calculate the tumor formation rate.</p><p><b>RESULTS</b>The tumor formation rates were 100% in all the 3 groups. No significant difference was found in the tumor formation time among the F10 over-expression, F10-silenced and control groups (6.2 ± 0.78 vs 7 ± 2.49 vs 6.3 ± 0.67 days; F=0.781, P=0.468). A significantly greater tumor growth rate was noted in the F10 over-expression group compared with the other two groups (P<0.05), and the growth rate was significantly slower in F10-silenced group than in the control group (P<0.05). The subcutaneous tumor weight at 5 weeks after JEG-3 cell injection differed significantly among F10 over-expression, F10-silenced and control groups (571.1 ± 221.10 vs 136.2 ± 66.25 vs 354.5 ± 116.23 mg; F=21.199, P=0.000).</p><p><b>CONCLUSION</b>F10 gene plays a role in the regulation of choriocarcinoma JEG-3 cell proliferation and might enhance its tumorigenicity in nude mice.</p>

Expression and identification of recombinant protein of the new gene F10 which is associated with the path-ogenesis of hydatidiform mole / 医学研究生学报

Objective F10 is a new gene cloned from hydatidiform moles .Our previous study showed that the F10 gene might be associated with the invasion of trophoblast cells .This study was to construct a prokaryotic plasmid expressing the recombinant protein of F10 and isolate, purify, and identify it in order to further investigate the function of the F10 gene. Methods The coding sequence of the F10 gene was amplified from the human tissue by RT-PCR and cloned into the vector pET 28a.The plasmid was ex-pressed in the E.coli BL21(DE3) to obtain the F10 recombinant protein.The expressed F10 recombinant protein was purified with the chromatographic column and verified by sodium dodecyl sulfate polyacrylamide gel electropheresis ( SDS-PAGE) . Results The pro-karyotic plasmid expressing the F 10 recombinant protein was successfully constructed and highly purified F 10 recombinant protein was obtained. Conclusion The F10 recombinant protein was expressed and highly purified , which could be used as an antigen for the preparation of the F10 monoclonal antibody .

Fall risk estimation for senile patients / 中华物理医学与康复杂志

Objective To investigate the discriminant validity and relativity of three scales applied in fall risk estimation for senile patients.Methods The timed up and go test (TUGT),the Morse Fall Scale (MFS) and the Berg Balance Scale (BBS) were used by two trained testers to evaluate the fall risk of 161 senile in-patients.The patients were divided into a falling group and a no-fall group based on their history of falling in the previous one year.Student's t-test was applied to compare the discriminant validity of the TUGT,MFS and BBS.Spearman's correlation coefficient was calculated to analyze the correlation among the three scales.Results The scores of patients in the falling group on the three scales were significantly different from those of the no-falls group.The correlation coefficients among the three scales were in the range 0.680-0.888.Conclusion The TUGT,MFS and BBS all showed high sensitivity and good discrimination in fall risks estimation for senile patients.The results with the three scales were highly correlated.Because the emphasis of these three scales is different,a suitable scale should be selected in clinical practice according to the characteristics of the senile patient.

Effect of human chorionic gonadotrophin on TGF-?_3 expression in JEG-3 cell line / 中国免疫学杂志

Objective:To explore whether human chorioic gonadotrophin(hCG) might change the invasiveness of trophoblast by regulating the prodution of TGF-?3. Methods: The effect of hCG on TGF-?3 mRNA expression in JEG-3 cells was investigated by employing the semi-quantiative method of reverse transcription polymerase chain reaction(RT-TCR). Results:TGF-?3 was shown to be expressed in JEG-3 cells. After incubated with 0,50,500,5 000,25 000 mU/ml hCG for 48 hours respectively, the expression of TGF-?3 in JEG-3 cells increased gradually with the concentration of hCG increased. Additionally,TGF-?3 mRNA expression in JEG-3 cells incubated with 25 000 mU/ml hCG experienced a significant induction at 30 h point which was followed by a less significant decrease.Conclusion:The autocrine of TGF-?3 might be involved in the effect of hCG on trophoblast or trophoblast-derived choriocarcinoma cell invasiveness.

Macrophage colony-stimulating factor protects RAW264.7 cells from oxidative injury through the induction of MnSOD expression / 基础医学与临床

To show the protective effect of macrophage colony-stimulating factor (M-CSF) on macrophages under oxidative stress,we investigated the effect of M-CSF on RAW264.7 cells incubated with tert-butyl hydroperoxide (tbOOH) using L929 cell conditioned medium (L929-CM) as the source of M-CSF.The results showed that M-CSF could alleviate the tbOOH- induced oxidative injury to RAW264.7 cells.We also found that M-CSF could improve superoxide dismutase (SOD) activity in the cells.MnSOD mRNA expression was also shown to be increased by RT-PCR technique,and the induction could be blocked by actinomycin D.So,we concluded that M-CSF could protect RAW264.7 cells from oxidative injury through inducing MnSOD expression.

Effect of F10 gene on activities of transcriptional factor NF-?B and AP1 / 中国病理生理杂志

AIM:To investigate the influence of F10 on the activities of transcription factor NF-?B and AP1 in A549 cells.METHODS:The luciferase report plasmids of NF-?B-Luc,AP1-Luc and F10 gene were introduced into A549 cells and the luciferase activity was detected.The DNA binding activities of AP1 and NF-?B in the cells were measured by the electrophoretic mobility shift assay(EMSA).RESULTS:The luciferase activity in F10+ transfection group decreased 30% and increased 2-fold respectively 48 h after transfected with the luciferase report plasmid of NF-?B-Luc and AP1-Luc in A549 cells.The DNA binding activity decreased 49% and increased 23%,respectively.CONCLUSION:F10 gene up-regulates the transcription activity of AP1 and down-regulates the NF-?B in A549 cells.

Detection of metabolism-associated genes expression in preeclamptic placenta by employing gene microarray / 中国病理生理杂志

AIM: To study the change of some metabolism-associated genes expression in preeclamptic placenta. METHODS: The mRNA expression levels of metabolism-associated genes in preeclamptic or normal placenta were detected by employing complementary DNA microarray representing over 220 human hormone-associated genes. RESULTS: Many redox metabolism-related genes (GenBank: U78168, X16699, D32143, AL035079, AF069668, X53463, J03746, X02317, X91247, etc) were highly expressed in preeclamptic placenta compared with normal placenta. Additionally, the mRNA expression levels of some genes which were related to other metabolism such as hormone (GenBank: NM -001718, Z22535, M38180, M76665, X75252, J03258, U56725), were also highly expressed in preeclamptic placenta.CONCLUSION: The change of genes expression in placenta is associated with the change of metabolism in patients sufferring from preeclampsia.

Effect of human chorionic gonadotrophin on vascular epithelial growth factor expression in JEG-3 cell line / 中国病理生理杂志

AIM: To uncover the effect of human chorionic gonadotrophin (hCG) on vascular epithelial growth factor (VEGF) expression in trophoblasts. METHODS: The semi-quantitative method of reversely transcription polymerase chain reaction (RT-PCR) was employed to investigate the expression of VEGF in trophoblast-derived JEG-3 cell line. RESULTS: VEGF 121 , VEGF 165 and VEGF 189 was shown to express in JEG-3 cells. After incubated with 0, 50, 500, 5 000 , 25 000 U/L hCG for 48 hours respectively, the expression of VEGF (including its 3 isoforms: VEGF 121 , VEGF 165 , VEGF 189 ) was induced gradually with the concentration of hCG increased. Furthermore, VEGF mRNA expression in JEG-3 cells incubated with 25 000 U/L hCG experienced a transient increase before a less significant decrease. CONCLUSION: The autocrine of VEGF might be involved in the effect of hCG on trophoblast or trophoblast-derived choriocarcinoma cell proliferation, migration and invasiveness.

Oxidant regulates macrophage colony-stimulating factor gene expression in RAW264.7 cells / 中国病理生理杂志

AIM:To find out the effect of oxidative stress on macrophage colony-stimulating factor (M-CSF) gene expression in macrophages.METHODS: The effect of tert-butyl hydroperoxide (tbOOH) on M-CSF expression in RAW264.7 cells was investigated by RT-PCR method.RESULTS: The results showed that tbOOH (1 5?10 -4 mol/L) could induce M-CSF mRNA expression in RAW264.7 cells; and the induction became greater with increase in tbOOH concentration. Furthermore, the induction of M-CSF in RAW264.7 cells by tbOOH could be attenuated by cycloheximide or acetovanilone added in the medium.CONCLUSION: The results indicated that tbOOH could transcriptionally induce M-CSF expression in RAW264.7 cells, and de novo protein synthesis and intracellular superoxide (O  2) production might be involved in the process.