OBJECTIVETo investigate the feasibility of plasmid nm23-h1 transfection on high metastatic potential adnoid cystic carcinoma (ACC-M) cell line mediated by cationic lipid.

METHODSACC-M cell were implanted in the maxillofacial region in each of 40 BALB/c nude mice. After the tumor growth to 1 cm in diameter, 0.1 ml Lipofctamine-nm23-hl plasmid complex were injected intratumorally in 10 mice, 3 days after the first injection, 10 mices injected for twice, 10 mice as plamid-blank control, another 10 mice were injected 0.2 ml complex, 2, 3, 7days after the injection, the mice were killed and the specimen for HE and immunohistological chemistry study.

RESULTSnm23-h1 expression initiated in the tumor cells 3 days after the complex injection, 7 days later, the expression level increased accompanying with extracellular matrix increase, twice injection and multiple channel injection would gain better nm23-h1 expression than once injection and single-channel injection respectively.

CONCLUSIONCationic lipid mediated nm23-h1 plamid transfecting adnoid cystic carcinoma can gain small range positive expression, but the results give little prospect for further clinical treatment in such a manner.

Animals ; Carcinoma, Adenoid Cystic ; Humans ; Lipids ; Mice ; Mice, Nude ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; Plasmids ; Transfection

OBJECTIVETo study the mechanism of sensitivity variation to cisplatin caused by nm23-H1.

METHODSThe samles was divided into two groups: Tca8113 group and Tca8113/nm23-H1 group. Using MTT and flow cytometer, the changes of cell mortality rate, apoptosis and mitochondrial membrane potential were detected. By VG PQ Excell, the changes of the intracellular platinum were detected.

RESULTSIn vitro the cell mortality rate and apoptosis were increased in Tca8113/nm23-H1 group, comparing with Tea8113 group. Mitochondrial membrane potential was decreased in Tca8113/nm23-H1 group. The intracellular platinum was increased significantly in Tca8113/nm23-H1 group. This effect could be inhibited by oubain which was an inhibitor of Na+/K+-ATP.

CONCLUSIONnm23-H1 can increase the sensitivity of cisplatin on Tca8113 cell line. The mitochondrial membrane potential was decreased by nm23-H1 so that intracellular platinum was increased and finally increased the apoptosis or necrosis.

Apoptosis ; Cell Line ; Cell Line, Tumor ; Cisplatin ; Humans ; In Vitro Techniques ; NM23 Nucleoside Diphosphate Kinases ; Transfection

OBJECTIVETo discuss the expression and clinical significance of VEGF-C and nm23-H1 in stage II and III colorectal carcinomas.

METHODSSP immunohistochemical staining was employed to determine the expression of vascular endothelial growth factor-C (VEGF-C) and nm23-H1 in the tumor tissues of 110 cases of stage II and III colorectal carcinomas and in the adjacent mucosal tissues of 53 cases as control, and analyze their correlation with cliniopathological features and prognosis.

RESULTSThe positive expression of VEGF-C in the carcinoma tissues was 71.8%, significantly higher than that in the adjacent mucosal tissues (22.6%, P < 0.001). The positive expression of nm23-H1 in the carcinoma tissues was 57.3%, significantly lower than that in the adjacent mucosal tissues (90.6%, P < 0.001). The expression of VEGF-C was significantly correlated with lymph node metastasis (P < 0.05), and the nm23-H1 expression was significantly correlated with lymph node metastasis and pathological type (P < 0.05). The expression of VEGF-C and nm23-H1 did not show a significant correlation with age, gender, primary tumor site, tumor size and depth of invasion (P > 0.05). The VEGF-C expression was negatively related with nm23-H1 expression in colorectal carcinoma (r = -0.361, P < 0.001). The median overall survival (MOS) and median disease free survival (MDFS) of 110 patients with colorectal carcinoma were 55 and 48 months, respectively. The colorectal patients with different VEGF-C and nm23-H1 expression showed significant differences in the 5-year OS rate and 5-year DFS rate (P < 0.001). The patients with negative VEGF-C expression and positive nm23-H1 expression had a better prognosis.

CONCLUSIONSThe joint detection of VEGF-C and nm23-H1 expression is very promising in prediction of the prognosis of patients with stage II and III colorectal carcinoma. However, whether it can be used as a marker in prognosis judgment needs further investigation.

Colorectal Neoplasms ; diagnosis ; metabolism ; Humans ; Lymphatic Metastasis ; diagnosis ; NM23 Nucleoside Diphosphate Kinases ; metabolism ; Prognosis ; Vascular Endothelial Growth Factor C ; metabolism

OBJECTIVETo study the effect of protein-cisplatin and nm23-H1 therapy on the tumor of nude mouse.

METHODSThe 15 BALB/C female mice were divided into three groups, control group, protein-cisplatin group and protein-cisplatin+nm23-H1 plasmid group. Tca8113 were injected into the mice subcutaneously with the concentration of 3.1 x 10(6) cells/mL. After two weeks, the mixture of lipofectin and nm23-H1 was injected around xenograft of nude mouse. After three days, the protein-cisplatin was injected around xenograft. The weight of mouse, the volume and the weight of xenograft were measured.

RESULTSThe weight of mouse was lightest in control group. The volume and weight of the transplanted tumor were lightest in nm23-H1 +protein-cisplatin group.

CONCLUSIONThe combination therapy of nm23-H1 and protein-cisplatin can effectively inhibites the growth of xenograft in nude mouse.

Animals ; Cisplatin ; Female ; Heterografts ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Transplantation ; Transfection

OBJECTIVETo construct a stable nm23-H1-knock-down cell model with K562 cell line and study its differentiation toward megakaryocyte.

METHODSEukaryotic expression vector pSilencer 4.1-CMV-sinm23 expressing siRNA targeting nm23-H1 was transfected into K562 cells with lipofectamine2000. Cells with stably nm23-H1 silence were screened out by G418. Real-time quantitative PCR, immunocytochemistry, western blot were used to confirm the nm23-H1-knock-down K562 model. Cell differentiation capacity was detected by NBT reduction assay. Surface antigen Gp IIb-IIIa (CD41) of knock-down cells treated with phorbol 12-myristate 13-acetate was analyzed by flow cytometry. Western blot was used to detect the ERK1/2 signal pathway after the stimulation of phorbol 12-myristate 13-acetate.

RESULTSEndogenous nm23-H1 was silenced by pSilencer 4.1-CMV-sinm23 and the silence efficiency was up to 75% and 70% in mRNA and protein levels respectively compared with the mock cells. Under phorbol 12-myristate 13-acetate treatment, the knock-down cells displayed a significantly increased differentiation ability toward megakaryocyte compared with control. The NBT reduction values were (0.31 +/- 0.07) and (0.23 +/- 0.05) respectively. Further results revealed that nm23-H1 gene regulating the megakaryocytic differentiation was due in part to the increased ERK1/2 phosphorylation.

CONCLUSIONSA stable nm23-H1-knock-down K562 cell model is successfully constructed. nm23-H1 involves in regulating the megakaryocytic differentiation of K562 cell line.

Cell Differentiation ; genetics ; Gene Knockdown Techniques ; Humans ; K562 Cells ; Megakaryocytes ; cytology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; RNA Interference

OBJECTIVETo investigate the relationship between expression of cell adhesion molecular CD44, epithelial cadherin (E-cad) and metastatic suppressor gene nm23-H1 as well as the clinicopathologic features including cell differentiation, invasion and metastasis of thyroid follicular-derived carcinoma.

METHODSForty two (42) thyroid follicular carcinomas (FTC) and 54 papillary carcinomas (PTC) were collected for studying the expression of CD44, E-cad and nm23-H1 using immunohistochemical staining.

RESULTSNeoplastic epithelium and infiltrating lymphocyte expressed CD44 in an intense plasma membrane pattern. CD44 expression rates in poorly differentiated FTC (80%) and PTC cases with metastasis (78%) were significantly higher than those of well-differentiated FTC cases (64%) and PTC without metastasis cases (59%) respectively. Thyroid carcinoma tissue was positive for E-cad and nm23-H1 in a cytoplasm pattern. Well-differentiated FTC presented a higher E-cad and nm23-H1 expression rate than poorly-differentiated FTC, but both had a lower expression rate than that of PTC (70% and 76%, P < 0.01). The expression rate and intensity of E-cad and nm23-H1 were lower and less in metastatic PTC than those in primary PTC. Expression rate of CD44 (72%) in thyroid follicular-derived carcinoma was higher than those of E-cad (54%, P < 0.01) and nm23-H1 (61%, P < 0.05). E-cad expression was adversely correlated with that of nm23-H1 (chi(2) = 15.75, P < 0.011, r = 0.522 2). There was a reverse relationship between expression of CD44 and E-cad or nm23 (P > 0.05).

CONCLUSIONSCell differentiation degree in FTC and metastasis in PTC have positive correlation with the expression of E-cad and nm23, but have a reverse correlation with the expression of CD44. There was a relationship between expression of CD44, E-cad, nm23 and the characteristics of the degree of differentiation, metastatic potential and the prognosis of thyroid follicular-derived carcinoma.

Adult ; Aged ; Cadherins ; analysis ; Carcinoma, Papillary, Follicular ; chemistry ; pathology ; Female ; Humans ; Hyaluronan Receptors ; analysis ; Immunohistochemistry ; Male ; Middle Aged ; Monomeric GTP-Binding Proteins ; analysis ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Invasiveness ; Nucleoside-Diphosphate Kinase ; Thyroid Neoplasms ; chemistry ; pathology ; Transcription Factors ; analysis

OBJECTIVETo transfect nm23-H1 into the BcaCD885 cell lines in order to get safe high-efficiency and low-toxicity, and to find out whether nm23-H1 could affect the invasion and metastases ability of BcaCD885 cell lines.

METHODSLipofect was used to transfect nm23-H1 into BcaCD885 cell lines; immunohistochemistry was used to detect the difference expression of nm23-H1 between transfected and non-transfected cell lines; then transwell-room and wash way were used to detect the difference of invasion and metastases ability between transfected and non-transfected cell lines.

RESULTSPCMV-NEO-BAM system gave the stability expression of nm23-H1; there was significant different NDPKA expression between transfected and non-transfected BcaCD885 cell lines; the invasion and metastases ability of transfected BcaCD885 cell lines decreased obviously.

CONCLUSIONnm23-H1 can inhibit the metastases of BcaCD885 cell lines significantly.

Cell Adhesion ; physiology ; Cell Movement ; physiology ; Genetic Vectors ; genetics ; Humans ; Monomeric GTP-Binding Proteins ; genetics ; metabolism ; Mouth Neoplasms ; genetics ; pathology ; physiopathology ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; Transcription Factors ; genetics ; metabolism ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo assess the significance of expression of sialylated carbohydrate antigens and nm23-H1 gene in metastasis and prognosis of breast cancer.

METHODSTissue specimens from 102 cases of primary breast cancer were stained with antibodies against sialyl Lewis A (SleA) and salyl Lewis X (SleX), and nm23-H1 proteins by immunohistochemical methods.

RESULTOf the 102 cases, the positive cases of SleA and SleX were 24.5% (25/102) and 59.89% (61/102),respectively; the reduced expression of nm23-H1 was showed in 37.3% (38/102) of the cases. The positive expression of SleX and the reduced expression of nm23-H1 gene were significantly associated with lymph node involvement. Among the 100 patients who underwent curative surgery, the disease-free survival rate was significantly correlated with nm23-H1 and SleX expression, respectively,but not with SleA expression. In multivariate analysis using Cox regression model, combination assay of nm23 H1 and SleX expression emerged as independent prognostic factors.

CONCLUSIONThese results suggest that nm23-H1 gene and SleX may be involved in the metastatic process in human breast cancer, and immunohistochemical detection of SleX and nm23-H1 may be used as a biologic marker of prognosis.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; chemistry ; genetics ; mortality ; Female ; Gangliosides ; analysis ; Humans ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; genetics ; Oligosaccharides ; analysis ; Prognosis ; Survival Rate

OBJECTIVETo identify molecular markers of lung squamous cell carcinoma by cDNA microarray technique.

METHODScDNA expression profiles were examined by microarrays of 6 surgical specimens of stage I lung squamous cell carcinomas. Those genes, either up-regulated or down-regulated in every specimen studied, were identified. The expression levels of nm23 and BRCA2 by the squamous cell carcinoma of the lung were further examined by immunohistochemical techniques.

RESULTSA total of 107 genes were identified, of which 26 were up-regulated and 81 were down-regulated in all six specimens. Immunohistochemical staining showed that, compared with normal lung tissues, the intensity of nm23 expression by the squamous cell carcinoma of lung was significantly increased while that of BRCA-2 was decreased.

CONCLUSIONcDNA microarrays can be used to identify gene expression profile of lung cancer, some of which may be used as markers of lung squamous cell carcinoma.

BRCA2 Protein ; metabolism ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Gene Expression Profiling ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Male ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; metabolism ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo study the expression of p16 and nm23 genes in salivary gland tumors and the relation of P16 and nm23 proteins with fumorigenesis of salivary gland tumors.

METHODSExpression of P16 and nm23 proteins was examined by SABC immunohistochemical method in 39 cases of paraffin blocks of normal salivary gland tissues and salivary gland tumors.

RESULTSP16 and nm23 protein positive staining were mainly found in the cytoplasm and cytoblast of all salivary gland tissues. Positive rate of P16 protein expression was 76.9% (10/13) and 40.9% (9/22) in benign and malignant salivary gland tumors, respectively. There was significant difference between P16 protein expression of benign and malignant tumors by chi 2 test (P < 0.05). mm23 protein positive staining was found in 84.6% (11/13) and 45.5% (10/22) of benign and malignant tumors respectively. The expression of nm23 protein in benign and malignant tumors was significantly different (P < 0.05). There was no correlation of the expression of P16 and nm23 in salivary gland tumors was found (P > 0.05).

CONCLUSIONp16 and nm23 genes may play an important role in different sides in salivary gland tumorigenesis and the reduce of the expression of p16 and nm23 genes may contribute to the generation of malignant salivary gland tumors.

Adenoma, Pleomorphic ; genetics ; metabolism ; Carcinoma, Mucoepidermoid ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Humans ; Immunohistochemistry ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; Protein Biosynthesis ; Proteins ; genetics ; Salivary Gland Neoplasms ; genetics ; metabolism ; Salivary Glands ; metabolism