Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.

Purpose To investigate the effect of down-regulation of miR-92 a on the proliferation and angiogenesis of nonsmall cell lung cancer (NSCLC). Methods Human NSCLC cell A549 was divided into three groups: A549 group (non-transfected A549 cells), sc-siRNA group (A549 cells transfected with sc-siRNA) and miR-92a-siRNA group (A 5 4 9 cells trans-fected with miR-92a-siRNA). The relative expression level of miR-92 a, PTEN and vascular endothelial growth factor (VEGF) in A549 cells and human bronchial epithelial (HBE) cells were detected by RT-PCR and Western blot respectively. The proliferation ability of A549 cells in each group was detected by living cell count and crystal violet staining experiment. Results The relative expression of miR-92 a in A549 cells was significantly higher than that in HBE cells (P < 0.05), the expression level of PTEN protein in A549 cells was significantly lower than that in HBE cells (P < 0.05), and the expression level of VEGF protein was significantly higher than that in HBE cells (P < 0.05).In the miR-92a-siRNA group, the relative expression of miR-92 a decreased (P < 0.05), the expression level of PTEN protein in-creased (P < 0.05), and the expression level of VEGF protein decreased (P < 0.05). The expression levels of PI3 K and Akt in miR-92a-siRNA group decreased (P < 0.05). the number of cells and cell proliferation ability in miR-92a-siRNA group reduced. Conclusion The expression of miR-92 a in NSCLC A549 cells is up-regulated, miR-92 a gene silencing can significantly inhibit cell proliferation and inhibit cell angiogenesis, PTEN and VEGF related PI3K/Akt signaling pathways may play an important role in this process.

OBJECTIVETo detect the expression of RhoC and Rho kinase 1 (ROCK-1) in prostate carcinoma, and explore the possible mechanism of RhoC/ROCK-1 in the pathogenesis of prostate carcinoma.

METHODSTissue specimens from 73 patients with prostate carcinoma and corresponding paracancerous tissues were obtained by prostate cancer biopsy or radical prostatectomy. The expression of RhoC/ROCK-1 mRNA was detected by RT-PCR. Western blot and immunohistochemistry were performed to dertect the expression of RhoC/ROCK-1 protein. Eukaryotic expression plasmids of RhoC were constructed and transfected into PC-3M-2B4 cells. p-MAPK and p-Akt were detected by Western bolt.

RESULTSThe expression levels of RhoC and ROCK-1 mRNA in the prostate carcinomas were significantly higher than those in corresponding paracancerous tissues [72.6% (53/73) vs. 34.2% (25/73); 68.5% (50/73) vs. 38.4% (28/73), P < 0.01], respectively. The results indicated that RhoC/ROCK-1 mRNA expression had no significant correlation with Gleason grade. However, the expression of RhoC/ROCK-1 mRNA showed a significant positive correlation with distant metastasis. The RhoC/ROCK-1 protein expression in prostate cancer was also higher than corresponding paracancerous tissues, and showed a significant positive correlation with p-MAPK and p-Akt expression levels. In addition, p-MAPK and p-Akt expression levels were up-regulated in the transcripts.

CONCLUSIONExpression levels of RhoC and ROCK-1 in prostate carcinoma are higher than those in corresponding paracancerous tissues, showing a significant positive correlation with distant metastasis. RhoC/ROCK-1 may be involved in the development, invasion and metastasis of prostate carcinoma.

Bone Neoplasms ; metabolism ; secondary ; Cell Line, Tumor ; Humans ; Male ; Mitogen-Activated Protein Kinases ; metabolism ; Neoplasm Grading ; Neoplasm Staging ; Phosphorylation ; Prostatectomy ; Prostatic Neoplasms ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; metabolism ; Transfection ; Up-Regulation ; rho GTP-Binding Proteins ; genetics ; metabolism ; rho-Associated Kinases ; genetics ; metabolism ; rhoC GTP-Binding Protein

OBJECTIVETo observe the inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adhesion ability in vitro of ovarian cancer cells, and to investigate the mechanisms of action.

METHODSMMP-9 antisense oligonucleotides were transfected by lipofectinmin into ovarian cancer cell line HO-8910PM cells expressing MMP-9 induced with fibronectin. RT-PCR, Western blot and gelatin zymography were used to detected MMP-9 expression of mRNA and protein and enzymatic activity. The ability of invasion and migration of ovarian cancer cells was assayed in Transwell cell culture chamber. Cell adhersion assay was carried out in a microculture well pre-coated with fibronectin.

RESULTSMMP-9 expressions of mRNA and protein were significantly decreased in the antisense-transfected cells. Comparing with the control group, the inhibition rate was 34. 8% and 42. 5% , respectively (P <0. 05). Its gelatin enzymatic activity was inhibited. Matrigel invasion assay and Transwell migration assay revealed markedly reduction in invasion and migration for the antisense group. The inhibition rates were 22. 4% and 24. 8% , respectively. The adhesion ability was also reduced. The inhibition rates were 49. 8% and 38. 3% at 60 min and 90 min, respectively.

CONCLUSIONMMP-9 down-regulation can significantly inhibit the ability of invasion and attachment of ovarian cells in vitro. MMP-9 may play an important role in invasion and metastasis of ovarian cells and potentially be a molecular target of blocking invasion and metastasis of ovarian cancer.

Blotting, Western ; Cell Adhesion ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Down-Regulation ; Female ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Oligodeoxyribonucleotides, Antisense ; genetics ; Ovarian Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective To investigate the effect of Tetramethylpyrazin ( TMP) combined with cisplatin( DDP) on expression of a disintegrin and metalloproteinase with thrombospondin typeⅠmotif ( ADAMTS1 ) and vascular endothelial growth factor ( VEGF ) of Lewis lung cancer rats.Methods The Lewis lung carcinoma model of c57 bl/6 were produced by intraperitoneal injection of Lewis lung cancer cells in right axillary, per 0.2 mL.Then the 56 tumor-bearing mice were randomized into 4 groups: model group ( 0.9%NaCl, 0.2 mL ) , TMP group ( 100 mg· kg-1, 0.2 mL),DDP group(2 mg· kg -1, 0.2 mL),TMP+DDP group( doses as above, n=14).Different administration were served from day 5 transplantation and all mice were sacrificed after 19 days.The weight and volume of transplanted tumors were recorded.And the expressions of VEGF and ADAMTS1 were detected by immunhistochem-isty.Results Compared with model group, the growth of transplanted tumors in TMP group, DDP group, TMP+DDP group were significantly inhibited.The weight of tumor body in the three groups were markedly lower than that of model group.The inhibitory effect of TMP combined with DDP on tumor growth was enhanced com-pared with TMP or DDP alone treatment.The expression levels of VEGF were significantly lower but ADAMTS1 was sig-nificantly higher in TMP group,DDP group,TMP+DDP group than that in model group ( P<0.05 ).Pearson correla-ted analysis showed that the expression VEGE and ADAMTS1 negatively correlated(P<0.05).Conclusion TMP has synergetic inhibitory effect with DDP on transplanted tumor growth of Lewis lung carcinoma in mice.The action mecha-nism may be due to down-regulate the VEGF expression and up-regulate the ADAMTS1 expression,inhibiting tumor angiogenesis.

Objective To investigate the inhibitory effect of Tetrame-thylpyrazine ( TMP ) in combination with cisplatin on the tumor growth and angiogenesis in mouse Lewis lung cancer .Methods Lewis lung carcinoma cells were inoculated subcutaneously into right axillary of C57BL/6 mice, and the mice were randomly divided into 4 groups:model group, TMP group,cisplatin ( DDP) group and combination group ( TMP+DDP).From day 7 after inoculation, all the mice were given different treatments for 14 days, and change of the xenografted tumor volumes was monitored.All mice were sacrificed on day 21.Subcutaneous tumors were processed for tumor weight , and tumor growth inhibition rate and tumor necrosis rate of each group were calculated .The expression level of Kruppel -like factor 4 ( KLF4 ) , vascular endothelial growth factor ( VEGF ) were measured by Western blot . The microvascular density ( MVD) was determined by CD 105 staining.Results The tumor growth was suppressed in TMP group , cisplatin group and combination group .The weights of tumor were markedly decreased in TMP group , cisplatin group and combination group compared with model group ( P<0.05 ) .The combination group showed significant enhancement in anti -tumor efficacy than TMP group and DDP group (62.48%vs 35.58% vs 40.41%,P<0.01).Compared with DDP group,the tumor necrosis rate in combination groupwas significantly higher [ ( 24.64 ±11.96 )% vs ( 31.76 ±15.20 )%, P<0.05 ] , but not remarkably in TMP group [(15.58 ±3.19)%,P>0.05].The expression levels of vascular endothelialgrowth factor and microvessel density were significantly lower ,Kruppel-like factor 4 was significantly higher in TMP group , cisplatin group and combination group than that in model group ( P<0.05 ) .Meanwhile , there was no significant difference between cisplatin group and tetramethylpyrazine group ( P>0.05 ).Conclusion TMP has synergetic inhibitory effect with DDP on the growth and angiogenesis of Lewis lung carcinoma effectively by inhibiting expression of VEGF , enhancing expression of KLF4.

AIMTo explore the time-dependent changes of endogenous hydrogen sulfide system at the early stage of pulmonary hypertension induced by high pulmonary flow in rats.

METHODSEighty male SD rats, whose weight ranged 140 - 160 g, were randomly divided into control group (n = 40) and shunt group (n = 40). Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of high pulmonary flow. After 1 d, 3 d, 1 week, 4 week and 8 weeks of experiment, systolic pulmonary artery pressure (SPAP) of each rat, the H2S of rat lung tissue and CSEmRNA of rat lung tissue were evaluated, respectively.

RESULTSSPAP increased significantly as compared with those in control group in 1 week and 8 weeks of experiment. In contrast to control group, the H2S of rat lung tissue increased significantly on 3 d and in 4 weeks, respectively. Meanwhile, in contrast to control group, relative amount of CSE mRNA of lung tissues elevated significantly on 3 d and in 4 weeks, respectively. Moreover, SPAP and the H2S of rat lung tissue, the CSE mRNA of rat lung tissue correlated negatively in 1 week, 4 weeks and 8 weeks of experiment.

CONCLUSIONAnimal model of rats with high pulmonary blood flow exhibited pulmonary hypertension. Lung tissue H2S and CSE mRNA of rats exhibited double peaks within 8 weeks. These results revealed that endogenous H2S system might be relevant with the development of pulmonary hypertension induced by high pulmonary blood flow, and probably, it played a protective role in the regulation of pulmonary hypertension, especially, at its early stage.

Animals ; Hydrogen Sulfide ; metabolism ; Hypertension, Pulmonary ; metabolism ; physiopathology ; Lung ; blood supply ; metabolism ; physiopathology ; Male ; Pulmonary Artery ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the clinical value of HER2 overexpression in breast cancer and its prognostic implication in patients with lymph node negative breast carcinoma.

METHODSThe following electronic database were extracted using appropriate inclusive and exclusive standards: Cochrane library, PUBMED, Embase (1984 - 2003), OVID, CMCC and CNKI. Excel and RevMan 4.2 were used for statistical analysis.

RESULTSFifty-six articles were extracted to calculate the positive rate of HER2 overexpression. The pooled positive rate was 23.14% [19.54%, 26.73%], with positive immunohistochemistry (IHC) rate of 23.13% [19.49%, 26.77%] and positive FISH rate of 20.90% [15.54%, 26.25%]. Seven articles were used to evaluate prognostic predication of HER2 expression. It was concluded that in patients with lymph node negative breast carcinoma, HER2 overexpression (both IHC and FISH) independently predicted a poor prognosis based on disease-free survival (DFS) and overall survival (OS) with a P < 0.05. For DFS, the pooled RR was 1.38 [1.07, 1.80] with 1.16 [1.02, 1.31] for IHC and 1.98 [1.56, 2.52] for FISH. For OS, the pooled RR was 1.58 [1.16, 2.14] with 1.37 [1.14 to 1.64] for IHC and 2.33 [1.45 to 3.75] for FISH. HER2 overexpression effectively predicted DFS/OS of patients without adjuvant therapy and OS of patients with the therapy, but not for DFS, with the pooled RR of 1.46 [1.02, 2.09] and 1.11 [0.95, 1.31] for DFS, respectively and the pooled RR of 1.93 [1.44 to 2.58] and 1.25 [1.01, 1.56] for OS, respectively.

CONCLUSIONSIn patients with lymph node negative breast carcinoma, the positive rate of HER2 overexpression is 23.14%. HER2 overexpression indicates a poor prognosis and adjuvant therapy after surgery should be recommended.

Breast Neoplasms ; genetics ; metabolism ; pathology ; therapy ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Genes, erbB-2 ; Humans ; Lymph Nodes ; pathology ; Mastectomy ; Prognosis ; Receptor, ErbB-2 ; metabolism ; Survival Rate

OBJECTIVETo evaluate the relationship between combined multigene detection and response to adjuvant chemotherapy and prognosis in early-stage non-small cell lung cancer (NSCLC).

METHODSTissue microarray was prepared from samples of 86 cases of early-stage NSCLC who received adjuvant chemotherapy after radical surgery. The expressions of caspase-3, Fas, bax, bcl-2, survivin, PCNA, Ki67, MGMT, p53, p63, p73, p16, p27, VEGF, nm23, P-gp, MRP, LRP, GST-pi, Topo II, c-myc, cyclin-D1, Her-2, Cox-2, Ku70, Ku80, DNA-PKcs, ERCC1, MSH2, BCRP proteins were detected using immunohistochemical two-step method.

RESULTSThe positive rate of the 30 genes in lung cancer tissue were 27.9% - 91.9%, respectively. By univariate analysis, the expression of 8 genes was shown to be related with SCLC adjuvant chemotherapy. The cases with higher expression of survivin, P-gp, LRP, Ki67, p53, ERCC1 and lower expression of bax,VEGF had worse prognosis. By logistic regression analysis, the ERCC1, survivin, bax and VEGF were a marker group. Multivariate analysis showed the predict value of the response to adjuvant chemotherapy in early-stage NSCLC was 96.5%.

CONCLUSIONSurvivin, ERCC1, bax and VEGF are an ideal marker group to predict the effect of adjuvant chemotherapy in early-stage NSCLC.

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; pathology ; surgery ; Chemotherapy, Adjuvant ; DNA-Binding Proteins ; metabolism ; Endonucleases ; metabolism ; Female ; Follow-Up Studies ; Humans ; Inhibitor of Apoptosis Proteins ; Logistic Models ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Male ; Microtubule-Associated Proteins ; metabolism ; Middle Aged ; Neoplasm Staging ; Survival Rate ; Tissue Array Analysis ; Vascular Endothelial Growth Factor A ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the constitutive characteristics and the change trend of gynecologic malignant tumors in hospitalized patients in Guangxi Zhuang Autonomous Region over the recent 20 years.Methods Clinical data of 8009 in-patients who suffered from gynecologic malignant tumors in 23 hospitals from 1985 to 2004 in Guangxi Zhuang Autonomous Region were analyzed,with respect to the tumor types and change trend.Results(1)The leading 4 types of malignant tumors were cervical cancers, ovarian cancers,endometrial cancers,and malignant trophoblastic tumors according to the constitutive ratios of the tumors.The constitutive ratio of cervical cancer patients rose year by year,from 17.48% during the 1985-1989 period to 49.25% during the 2000-2004 period(P0.05).(2)The occurring age of cervical cancers became younger obviously,from≥60 years old dropped to