[Expressions of NF-κBp65 and IκBα in gestational trophoblastic disease and clinical significance].

AIM: To investigate the roles of nuclear factor κB p65 (NF-κBp65) and inhibitor of nuclear factor κB α (IκBα) in the development and metastasis of gestational trophoblastic neoplasia (GTN) by analyzing the expressions of NF-κBp65 and IκBα in normal early pregnancy villi and gestational trophoblastic diseases, and to reveal the relationship of NF-κBp65 and IκBα with age and clinical stage of GTN patients. METHODS: The expressions of NF-κBp65 and IκBα were detected by immunohistochemistry in normal pregnancy villi (20 cases), hydatidiform moles (HM, 30 cases), invasive hydatidiform moles (IHM, 13 cases) and chorionic carcinomas (CCA, 15 cases). RESULTS: NF-κBp65 expression was statistically different (P<0.05) between normal pregnancy villi and IHM (P=0.013), normal pregnancy villi and CCA(P=0.018), HM and IHM(P=0.026), HM and CCA (P=0.035). Differences in IκBα expression were statistically significant between normal pregnancy villi and IHM, normal pregnancy villi and CCA, HM and IHM, HM and CCA (P<0.01). The expressions of NF-κBp65 and IκBα were correlated to clinical stage (P=0.043, 0.042, P<0.05), but not to patients' ages. Spearman correlation analysis revealed that there was a negative association between the protein expressions of NF-κBp65 and IκBα in GTN (r=-0.403, P=0.034, P<0.05). CONCLUSION: Up-regulated expression of NF-κBp65 and down-regulated expression of IκBα may be related to the development, invasion and metastasis of GTN. The expressions of NF-κBp65 and IκBα are negatively correlated in gestational trophoblastic tumor tissues.

[Study of the bacterial community structure of microbiota in bacterial vaginosis using amplified ribosomal DNA restriction analysis].

OBJECTIVE: To study the bacterial community structure of the microbiota in the vaginal fluid from patients with bacterial vaginosis. METHODS: The composition of bacteria in the samples of vaginal fluid from 3 patients with bacterial vaginosis and 1 normal premenopausal control was investigated by amplified ribosomal DNA restriction analysis(ARDRA). RESULTS: Lactobacillus species were the predominant bacteria in the woman without bacterial vaginosis. Bacterial vaginosis was associated with higher concentrations of a variety of bacterial groups. Women with bacterial vaginosis had greater bacterial diversity, with 31 to 37 OTUs operational taxonomic units detected per sample. The species associated with bacterial vaginosis were Leptotrichia, Prevotella sp. and Megasphaera including several species with no close cultivated relatives. CONCLUSIONS: Women with bacterial vaginosis have complex vaginal infections with many newly recognized species. ARDRA allows rapid analysis of the diversity of microorganisms in the vagina, and is capable of identifying potentially pathogenic bacteria that can not be identified by general culture.

[Expression of caveolin-1 and the invasion of choriocarcinoma].

OBJECTIVE: To investigate the association between the expression of caveolin-1(CAV-1) and the invasion of choriocarcinoma, and to explore the effect of CAV-1 small interfering RNA(siRNA) on the invasion of choriocarcinoma cell line JEG-3. METHODS: (1) Matrigel invasion assay and 3-(4,4)-dimethylthiahiazo (-z-yl)-3,5-di-phenytetrazoliumormide (MTT) assay were used to examine the difference in invasion and proliferation ability between JEG-3 cells and JAR cells;(2) Expression of caveolin-1 gene in the human chorionic villi tissues and chorionicnoma cell lines (JEG-3 cells and JAR cells) were detected by semi-quantitative RT-PCR. (3) The effect of CAV-1 siRNA transfection on the expression of CAV-1 mRNA, and the invasion and proliferation ability of JEG-3 cells were measured by RT-PCR, Matrigel invasion assay and MTT assay. RESULTS: (1) The invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P<0.05), but the difference in proliferation ability between JAR and JEG-3 was not obvious (P>0.05);(2) The expression of caveolin-1 gene in chorionicnoma cell lines was significantly stronger than that in the human normal chorion(P<0.05), and the expression of caveolin-1 gene in JEG-3 cells was stronger than that in the JAR cells (P<0.05). The data suggested that there was significantly positive correlation between caveolin-1 and the invasiveness of chorionicnoma cells (r=0.086,P<0.05);(3) CAV-1 siRNA could knock-out the expression of CAV-1 mRNA, and inhibit the invasion and proliferation ability of chorionicnoma cells. CONCLUSION: CAV-1 can promote the invasion ability of chorionicnoma cells. CAV-1 siRNA can inhibit the invasion and proliferation ability of chorionicnoma cells.

[Correlation between expression of heparanase and invasion of choriocarcinoma].

OBJECTIVE: To investigate the association between the expression of heparanase (Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues. METHODS: (1) Matrigel invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells. (2) Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines (JEG-3 cells and JAR cells) were detected by immunocytochemistry and western blot. RESULTS: (1) The invasive cell number was significantly larger in JEG-3 cells than in JAR cells (191 +/- 17 vs 106 +/- 13, P < 0.05). (2) Hpa protein mainly located in cytoplasm by immunocytochemistry. (3) Hpa protein expression was stronger in JEG-3 cells than in the JAR cells (1.560 +/- 0.180 vs 0.610 +/- 0.170, P < 0.05); the Hpa protein expression was significantly stronger in choriocarcinoma cell lines than in human chorionic villous tissues (0.190 +/- 0.008) by western blot (P < 0.05). (4) The data suggested that there were significantly positive correlations between Hpa and the invasiveness of choriocarcinoma cells (r = 0.89, P < 0.05). CONCLUSIONS: (1) Hpa protein expression is significantly stronger in choriocarcinoma cell lines than in the human chorionic villous tissues. (2) Activation of Hpa enhances the invasion capability of choriocarcinoma. (3) Overexpression of Hpa may be related to the oncogenesis of choriocarcinoma and Hpa may play an important role in invasion of choriocarcinoma.

[Expression of heparanase and angiopoietin-2 mRNA in endometriosis].

OBJECTIVE: To investigate the relationship between heparanase (Hpa) and angiopoietin-2 (Ang-2) gene expression and the development of endometriosis (EM). METHODS: Expression of Hpa gene and Ang-2 mRNA in the eutopic and ectopic endometrium collected from 86 patients with endometriosis (EM group) and the normal endometrium from 30 women without endometriosis (control group) was determined by RT-PCR. RESULTS: (1) Hpa gene expression was detected in 53 ectopic endometrium and 47 eutopic endometrium specimens in the EM group and 8 normal endometrium specimens in the control group. There was no significant difference in Hpa gene expression between ectopic and eutopic endometrium in the EM group (P > 0.05). However, the Hpa gene expression in normal endometrium in the control group was significantly lower than that in ectopic and eutopic endometrium in the EM group (P < 0.05). (2) Ang-2 gene expression was detected in 61 ectopic endometrium and 56 eutopic endometrium specimens in the EM group and 10 normal endometrium specimens in the control group. There was no significant difference in Ang-2 gene expression between ectopic and eutopic endometrium in the EM group (P > 0.05), however, the Ang-2 positive rate in normal endometrium in the control group was significantly lower than that in ectopic and eutopic endometrium in the EM group (P < 0.05). (3) The expression of Hpa gene in stage III-IV endometriosis was significantly higher than that in stage I-II endometriosis (P < 0.05). (4) The difference of Ang-2 mRNA expression between stage III-IV and stage I-II endometriosis was not significant (P > 0.05). (5) The expression of Hpa gene was shown to be significantly correlated to the expression of Ang-2 gene in ectopic endometrium specimens from patients with endometriosis (P < 0.05). CONCLUSION: Hpa and Ang-2 genes are highly expressed in both ectopic and eutopic endometrium of patients with endometriosis. Over-expression of these two genes may play a role in the development and progression of endometriosis.