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 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 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 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 explore nm23 gene mRNA expression and its clinical significance in acute leukemias (AML).

METHODSThe levels of nm23-H1 and nm23-H2 transcripts in 22 patients with acute myeloid leukemia (AML), 9 AML in complete remission (AML-CR), 12 acute lymphoblastic leukemia (ALL) and 4 chronic myeloid leukemia in chronic phase (CML-CP) were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSThe expression of nm23-H1 in AL especially in AML-M4 and AML-M5 was significantly higher than that in normal blood cells. An analysis of correlation between nm23 expression and clinicopathological parameters showed that increased nm23-H1 mRNA levels were associated with some poor-prognostic factors such as extramedullary infiltration, high white blood cell count (WBC), high lactate dehydrogenase (LDH) activity and high CD(7) expression, while inversely correlated with t(8; 21) and t(15; 17) which had a good-prognostic effect. The expression of nm23-H1 in AML patients in CR was significantly decreased compared with those untreated.

CONCLUSIONnm23-H1 was overexpressed in AL, especially in AML-M4 and AML-M5. High expression of nm23-H1 may be a poor prognostic factor.

Adult ; Female ; Gene Expression ; Humans ; Leukemia, Myeloid, Acute ; genetics ; pathology ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

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 regulation and effect of the expression of nm23-H1 gene in different processes of regional lymph node metastases of oral squamous cell carcinomas(OSCC).

METHODSBy immunohistochemical analysis in 200 paraffin-embedded tissues of OSCC and Western Blot in 9 fresh tissues of OSCC using a monoclonal antibody, the expression of nm23-H1/NDPK-A were detected in dividing groups accompanied with the clinical and pathological data of cervical lymph node metastases and modes of invasion.

RESULTSThe rates of negative expression of nm23-H1/NDPK-A had significant differences between metastatic cases(34/81) and non-metastatic cases(15/119). The nm23-H1/NDPK-A negative group showed higher frequency of lymph node metasteses (P < 0.01). In the different metastatic processes, There were significantly differential expressions of nm23-H1/NDPK-A among stage N0, N1 and N2(P < 0.01), distinct involved quantities of lymph nodes (P < 0.01), dissimilar metastatic involved levels(P < 0.05), definite modes of invasion(P < 0.01) and different cell differentiations(P < 0.01). There were much higher negative expressions in those cases of N1 stage(23/38), only one lymph node to be involved(23/39), metastasized to submandibular nodes or/and superior deep cervical nodes(17/28 and 12/29, respectively), IV c types of invasion(33/54) and poor differentiations(9/19).

CONCLUSIONThe results indicated that nm23-H1/NDPK-A played an more important role in switching the initial metastases formation than in influencing the later metastases spread because the negative expressions of nm23-H1/NDPK-A in solid tumors of OSCC would induce the formation of high metastatic cellular subpopulations. It was also confirmed that nm23-H1 gene might be a metastatic suppressor and may be useful in predicting the initial lymph node metastases in OSCC.

Carcinoma, Squamous Cell ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Lymphatic Metastasis ; Mouth Neoplasms ; genetics ; pathology ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; Protein Biosynthesis ; Proteins ; genetics