Objective To study the expression situation of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the endometrium of women with unexplained infertility and normal endometrium, and to explore the relationship between MMP-9/TIMP-1 expres-sion and unexplained infertility. Methods Immunohistochemieal assay(SP method) was employed to deteet the expression of MMP-9/TIMP-1 in 20 cases of impaired endometrium(unexplained infertility group,endometrial plantation window phase)and 20 cases of normal endometrium(heahhy control group). Results There were different levels of MMP-9/TIMP-1 expression in kytoplasms of glandular epicytes and stromal cells of all endometrial samples. The expression of MMP-9/TIMP-1 was signifi-cantly weaker in unexplained infertility group than that in healthy control group(P＜0.05). Conclusion Low expression of MMP-9/TIMP-1 in the endometrial plantation window phase may be one of impor-tant factors for unexplained infertility.

Aim To investigate the effects of triptolide on the proliferation and apoptosis of cell line A549 in human Lung Adenocarcinoma. Methods MTT, DNA fragmentation assay, fluorescence stain and flow cytometric analysis were carried out.Results Triptolide notably reduced the survival rate of A549 cells, and inhibited the proliferation of A549 cells. The 50 % inhibitory concentration (IC50) for 72 h was 75 nmol?L-1 and it arrested the cell cycle at S phase at the concentration of 14 nmol?L-1 (P

AIM: To observe the expression of HIF-1? mRNA, HIF-1?, VEGF and iNOS proteins and to investigate their relationship in h ypoxia-treated SW480 cells. METHODS: HIF-1?, VEGF and iNOS proteins were measured by immuno cytochemistry. Western-blot was used to detect HIF-1? protein. HIF-1? mRNA wa s measured by in situ hybridization. RESULTS: Under hypoxic condition, SW480 cells expressed proteins of HIF-1?, VEGF and iNOS more strongly than that under normoxia condition. How e ver, under hypoxia condition, these three proteins expressed weakly or negativel y when the cells treated with genistein, the inhibitor of HIF-1?. Expressions o f HIF-1? and VEGF proteins in cultured SW480 cells under hypoxic condition were completely or partially inhibited by the addition of SNP but the expression of iNOS was unaffected. Another NO donor NOC5, however, induced the expression of t hese three proteins. L-NAME, a non-specific inhibitor of NOS, inhibited the expr ession of HIF-1?, VEGF and iNOS. The levels of HIF-1? mRNA changed slightly i n different oxygen condition or addition of genistein, NO donor or iNOS inhibitor . CONCLUSIONS: Hypoxia induces the expression of HIF-1?, therefor e upregulates the production of VEGF and iNOS. During hypoxia, SNP inhibits but N OC5 promotes HIF-1? expression, indicating that different NO donor acts on the cells through different mechanisms.

OBJECTIVETo investigate the dynamic expression and its relation of gelatinase A (MMP-2), its natural inhibitor (TIMP-2) and DNA index (DI) changes during carcinogenesis, invasion and metastasis in Wistar rats.

METHODSSquamous cell carcinoma of lung was induced with 3-methylcholanthrene (MCA) and diethyinitrosamine (DEN) in iodized oil by left intra-bronchial instillation in 80 Wistar rats. Immrno histochemistay (IHC) and in situ hybridigation were used in the monitor of MMP-2, TIMP-2 proteins and mRNA expression during invasion and metastasis of lung cancer in these rats, DNA index (DI) value was measured by guantitatove image analysis on feulgen stained sections.

RESULTSAlong with the carcinogenis, the average poritive MNP-2 and TIMP-2 expressions increased, with positive rates of 8.5% - 85.7% and 6.4% - 35.7%. DI value also underwent the same changes (1.47 +/- 0.54) - (2.87 +/- 0.55). The difference of MMP-2 expression in carcinoma in situ versus early carcinoma and early carcinoma versus metastatic carcinoma are statistically significant (P < 0.05). Companing lung carcinome, the contrel group and non-cancerous lesions, the elevation of MNP-2 and TIMP-2 expressions were also sigmificant (P < 0.01). The DI elevation in carcinoma in situ and dysplasia were obviously significant (P < 0.05). Meanwhile a negative relation was noted in TINP-2 and MMP-2 expressions during carcinogenesis. There was a positive relation between MMP-2 expression and DNA poikiloidy (P < 0.01), which was related to the close relationship between MMP-2 and metastasis in advanced rat lung carcinoma (P < 0.05).

CONCLUSIONThe excess degradation and disruption of basement membranes by activated MMP-2 may be a key step in inducing lung cancer invasion and metastasis. The imbalance between MMP and TIMP may be a critical factor which affects biologic behavior of lung carcinogenesis, invasion and metastasis.

Alkylating Agents ; toxicity ; Animals ; Carcinoma, Squamous Cell ; chemically induced ; genetics ; pathology ; Diethylnitrosamine ; toxicity ; Female ; Gene Expression Regulation, Neoplastic ; Lung ; drug effects ; metabolism ; pathology ; Lung Neoplasms ; chemically induced ; genetics ; pathology ; Male ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Methylcholanthrene ; toxicity ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism

BACKGROUNDTo analyse the relationship between caspase-3 expression and cell proliferation, and to find molecular-biology markers to adjust canceration during rat lung squamous cell carcinogenesis.

METHODSThe 3-methylcholanthrene(MCA) and diethyinitrosamine (DEN) were used to induce lung squamous cell carcinoma by intra-left lobar-bronchial instillation in 50 Wistar rats, and 10 normal rats as controls. Expression of caspase-3 and PCNA were evaluated by immunohistochemistry (IHC).

RESULTSCaspase-3 protein positive rate was 44.12% in 34 rat lung squamous cell carcinomas, and positive coefficient value was 1.38±0.95, which were significantly lower than that of normal bronchial epithelium (P=0.007, P < 0.01) and premalignant lesions (P < 0.05, P < 0.05). The mean PCNA-labeling indexes (PCNA-LI) of normal rat bronchial epithelium, premalignant lesions and lung cancer were 14.10±5.02, 28.13±8.72 and 41.88±14.24 (P < 0.05), respectively. There was a negative correlation between caspase 3 and PCNA LI (r=-0.730 6, P < 0.01).

CONCLUSIONSLoss expression of caspase-3 may promote tumor cell growth, and it may be important in rat lung squamous cell carcinogenesis. Detection of caspase-3 and PCNA proteins can be regarded as major markers in the diagnosis of lung canceration.

OBJECTIVETo investigate the roles of p53 and K-ras gene in carcinogenesis and development of the lung carcinoma induced by 3-methylcholanthrene (MCA) and diethylinitrosamine (DEN) in Wistar rats, and to elucidate the relationships between the protein expression and gene mutation of p53 and K-ras.

METHODSMicrodissection was used to obtain pure cell populations of each phase in the carcinogenesis and development of lung carcinoma induced by MCA and DEN. DNA of the microdissected cell populations was extracted and used to analyze the mutations of p53 exons 5 approximately 8 and K-ras exons 1 approximately 2 by PCR-SSCP. The expressions of p53 and K-ras protein in each phase were detected by immunohistochemistry.

RESULTSNo mutation and protein expression of p53 and K-ras was found in the 30 cases with normal bronchial epithelium. Mutation of p53 was detected in 3.1% of 18 hyperplasia and 14 squamous metaplasia cases, 28.6% of 21 dysplasia, 30.0% of 12 carcinomas in situ, 51.2% of 43 infiltration carcinomas, 52.9% of 17 metastases. The positive immunostaining rate of p53 protein was 0, 42.9%, 50.0%, 60.5% and 64.7% respectively. K-ras mutation rate was 0, 4.8%, 8.3%, 9.3%, 11.8% respectively, while the overexpression rate of K-ras protein was 15.6%, 19.0%, 25.0%, 41.9%, 52.9% respectively. p53 protein expression was closely related with p53 mutation (P < 0.005, Pearson's R = 0.599 6). There was no relationship between the protein expression and gene mutation of K-ras (P > 0.500).

CONCLUSIONSp53 gene mutation and K-ras overexpression were early events in the carcinogenesis and development of rat lung carcinoma induced by MCA and DEN, while K-ras mutation does not play any important role.

Animals ; Genes, p53 ; Genes, ras ; Humans ; Immunohistochemistry ; Lung Neoplasms ; chemically induced ; chemistry ; genetics ; Mice ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Rats ; Tumor Suppressor Protein p53 ; analysis ; ras Proteins ; analysis