Nucleoside diphosphate kinase (Ndk) is ubiquitous and highly conserved multifunctional key enzyme in nucleotide metabolism. It generates nucleoside triphosphates (NTPs) by transfer of gamma-phosphates from nucleoside triphosphates such as ATP or GTP to nucleoside diphosphate. The formation of an autophosphorylated enzyme intermediate is involved in that mechanism. The phosphate is usually supplied by ATP and Ndk activity in different subcellular compartments. Ndk may regulate the crucial balance between ATP and GTP or other nucleoside triphosphates. Ndk is playing an important role in bacterial pathogenesis and emerging evidences recognize multiple roles of Ndk in host-microbe interaction. Here, I review some examples of the role of Ndk in intra- and extracellular microorganism.
Adenosine Triphosphate ; Guanosine Triphosphate ; Nucleoside-Diphosphate Kinase

OBJECTIVETo study the molecular mechanisms of nm23-H1 for regulating PKC signal pathway before and after transfection with nm23-H1 gene.

METHODSUsing Western-blot, Boyden-chamber, MTT and laser scanning confocal microscopy (LSCM) techniques to detect the distribution of PKC in cytosol and plasma membrane, changes of invasion and proliferation activity, PKC translocation status and changes of intracellular Ca(2+) concentration among different human pulmonary carcinoma cells with transfected or untransfected nm23-H1 gene, and changes of the three cell lines after treatment with Calphostin C, a PKC inhibitor.

RESULTS(1) The expression of PKCalpha, PKCbeta II on L9981 and L9981-pLXSN cell membrane, which was in activated status, was remarkably higher than those in L9981-nm23-H1 cell line (P < 0.001). The expression of PKCalpha, PKCbeta II in cytosol in L9981 and L9981-pLXSN cell lines, which was in inactivated status, was lower than those in L9981-nm23-H1 cell line (P < 0.001). It means that the PKC signal pathway was activated in L9981 and L9981-pLXSN cell lines. (2) PKCalpha and PKCbeta II mainly located in nuclei and perinuclear area in L9981 and L9981-pLXSN cells, which were in active status, and the Ca(2+) concentration in these cells was obviously higher than that in L9981-nm23-H1 cell line (P < 0.01). In L9981-nm23-H1 cell line, which was transfected with nm23-H1 gene, PKCalpha and PKCbeta II mainly located in soluble cytosolic section, in an inactive status. (3) The invasion and proliferation ability of L9981 and L9981-pLXSN lung cancer cells was higher than that of L9981-nm23-H1 cell line (P < 0.001). There was no statistically significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05). (4) After treated with PKC inhibitor Calphstin C, the expression of PKC and PKCbeta II in membrane in L9981 and L9981-pLXSN cell lines was down-regulated (P < 0.001), PKCalpha and PKCbeta II were mainly located in cytosolic area, mainly in an inactive status, and the Ca(2+) concentration was found to be decreased in all the three cell lines. The invasion and proliferation ability of the three lung cancer cell lines were obviously decreasing (P < 0.001). However, the invasion and proliferation ability of L9981-nm23-H1 lung cancer cell line was still lower than that of L9981 and L9981-pLXSN lung cancer cell lines (P < 0.001). There was also no significant difference between L9981 and L9981-pLXSN cell lines (P > 0.05).

CONCLUSIONThe results of this study suggest that nm23-H1 gene might inhibit the invasion and metastasis of lung cancer cells by down-regulating PKC signaling pathway. The Ca(2+) in cells might be involved in this process.

Calcium ; metabolism ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cell Proliferation ; drug effects ; Cytosol ; metabolism ; Down-Regulation ; Humans ; Lung Neoplasms ; enzymology ; metabolism ; pathology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; Naphthalenes ; pharmacology ; Neoplasm Invasiveness ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Protein Kinase C beta ; Protein Kinase C-alpha ; metabolism ; Signal Transduction ; Transfection

OBJECTIVETo study lymph node micrometastases (LNMM), expression of nm23-H(1), MMP(9), TIMP(2) proteins, and their relationship and clinical significance in patients with stage Dukes B colorectal cancer.

METHODSThirty patients with stage Dukes B colorectal cancer were studied. LNMM in these patients was detected by immunohistochemical anti-cytokeratin 20 (CK20) staining. The expression of nm23-H(1), MMP(9) and TIMP(2) proteins in primary tumors was examined by Strept-avidin-biotin complex method. Clinical-pathological data and survival of each patient were recorded and analyzed.

RESULTS(1) The positive dyeing of CK20 was observed in 26.7% for cases and in 7.8% for lymph nodes of 30 patients with stage Dukes B colorectal cancer. (2) Different expression of nm23-H(1) and MMP(9) proteins in the patients between stage Dukes B and stage Dukes CD was observed (P < 0.05). The decreased nm23-H(1) expression, and/or the increased MMP(9) expression in primary stage Dukes B tumors were significantly associated with LNMM (P < 0.05). Sensitivity and specificity for detection of LNMM by using nm23-H(1) or MMP(9) were respectively 62.5% and 81.8% or 75.0% and 69.8%. If by combining nm23-H(1) with MMP(9), specificity for detection of LNMM became 90.9%. The expression of TIMP(2) protein was not related with stage Dukes and LNMM. (3) The percent of tumor recurrence and/or metastasis for the stage Dukes B patients with LNMM was significantly higher than that for the patients without LNMM (P < 0.05), but the survival percent for the patients with LNMM was significantly lower than that for the patients without LNMM. The outcome for the patients with nm23-H(1) (-) LNMM (+) or MMP(9) (+) LNMM (+) was significantly worse than that for patients with nm23-H(1) (+) LNMM (-) or MMP(9) (+) LNMM (-) (P < 0.05).

CONCLUSIONSLNMM is detected by immunohistochemical anti-CK20 staining. The expression of nm23-H(1) and MMP(9) in primary stage Dukes B tumors was significantly associated with LNMM. The outcome in the LNMM patients with nm23-H(1) (-) and/or MMP(9) (+) were worse. Combining examination of CK20 for lymph nodes with expression of nm23-H(1) and MMP(9) for primary tumors is of important clinical significance for staging of Dukes, selection of adjuvant treatment and evaluation of prognosis in patients with colorectal cancer.

Colorectal Neoplasms ; metabolism ; pathology ; therapy ; Humans ; Keratins ; metabolism ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Matrix Metalloproteinase 9 ; metabolism ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Staging ; Nucleoside-Diphosphate Kinase ; metabolism ; Prognosis ; Tissue Inhibitor of Metalloproteinases ; metabolism

The metastasis-suppressing role of the nm23 gene in the metastatic spread of malignant tumor is still debated. We examined the nm23-H1 protein expression and gene mutation in non-Hodgkin's lymphomas to compare with the clinicopathologic parameters. The expression of nm23-H1 protein was immunohistochemically examined in 150 cases of non-Hodgkin's lymphomas; 85 diffuse large B cell lymphomas (DL-BCL), 18 marginal zone B cell lymphomas (MZL), 3 mantle cell lymphomas, 25 peripheral T cell lymphomas, not otherwise specified (TCLNOS), and 19 NK/T cell lymphomas (NK/T). Eighty-one cases (58 DLBCL, 6 MZL, 4 TCLNOS, and 13 NK/T) were studied for nm23-H1 gene mutation in exon 1 to 5. The high expression of nm23-H1 protein was associated with the high IPI score (p=0.019) and the low survival rate of the patients (p=0.0039). The gene mutation of nm23-H1 was detected in 10.3% of DLBCL and 30.7% of NK/T; but none in MZL and TCLNOS. The mutation was found in exon 1 in 5 cases, exon 2 in two cases, exon 4 in one case and both exon 1 and 2 in two cases. Our results suggest that the expression of nm23-H1 protein can be used as a poor prognostic marker in non-Hodgkin's lymphomas, and the mutational change of gene may operate in the lymphomagenesis.
Tissue Array Analysis ; Survival Analysis ; Prognosis ; Polymorphism, Single-Stranded Conformational ; Nucleoside-Diphosphate Kinase/*genetics/metabolism ; Mutation/*genetics ; Middle Aged ; Male ; Lymphoma, T-Cell/genetics/metabolism/pathology ; Lymphoma, Non-Hodgkin/genetics/metabolism/*pathology ; Lymphoma, Mantle-Cell/genetics/metabolism/pathology ; Lymphoma, Large-Cell, Diffuse/genetics/metabolism/pathology ; Lymphoma, B-Cell/genetics/metabolism/pathology ; Immunohistochemistry ; Humans ; Female ; DNA Mutational Analysis ; Base Sequence

Adult ; Aged ; Carcinoma, Non-Small-Cell Lung ; diagnostic imaging ; genetics ; mortality ; Female ; Genes, Tumor Suppressor ; Humans ; Lung Neoplasms ; diagnostic imaging ; genetics ; mortality ; Male ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; genetics ; Prognosis ; Tomography, X-Ray Computed

BACKGROUND: Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. METHOD: M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and IFN-gamma after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. RESULT: Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and IFN-gamma responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. CONCLUSION: Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.
Adenylate Kinase* ; Animals ; Clone Cells ; Enzyme-Linked Immunosorbent Assay ; Heat-Shock Proteins* ; HSP70 Heat-Shock Proteins* ; Immunoglobulin G ; Lung ; Mice* ; Mycobacterium bovis ; Mycobacterium tuberculosis* ; Mycobacterium* ; Nucleoside-Diphosphate Kinase* ; Polymerase Chain Reaction ; Recombinant Proteins* ; Spleen ; Tuberculosis ; Vaccination

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

OBJECTIVE: To investigate the expressions of Survivin protein and nm23 protein and the relationship among the expressions and axillary lymph node metastasis in breast cancer. METHODS: The expression of Survivin and nm23 in 80 cases of breast cancer tissues were detected by immunohistochemistry SP method, and their correlation with axillary lymph node metastasis and 5-year disease free survival (DFS) were analysed. RESULTS: Survivin protein positive expression rate was 68.75% (55/80) in breast cancer tissues, which had positive correlation with the axillary lymph nodes metastasis but negative correlation with 5 years FS (P < 0.05); nm23 protein expression had negative correlation with the axillary lymph nodes metastasis but positive to 5 years FS (P < 0.05). Survivin and nm23 proteins expression had no obvious correlation with the breast cancer pathology type, patient age and clinical stage (P > 0.05). CONCLUSION The anti-apoptosis effect of Survivin protein and the anti-metastasis effect of nm23 protein may be important in the occurrence and advancement of breast cancer, suggesting that it may be a new indicator of prognostic and judgement in breast cancer.
Adult ; Aged ; Axilla ; Biomarkers, Tumor ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mastectomy ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Proteins ; biosynthesis ; genetics ; Nucleoside-Diphosphate Kinase ; biosynthesis ; genetics ; Prognosis ; Retrospective Studies ; Survivin

OBJECTIVE: To explore the relationship between the expressions of PTEN and the metastasis of the gallbladder cancer. METHODS: The expression of PTEN and nm23 were detected by immunohistochemical staining in 32 cases of gallbladder cancer with metastasis and the staining intensity was scored semi-quantitatively, compared with the cases without metastasis. RESULTS: The intensity score of PTEN and nm23 in gallbladder cancer with metastasis was 8.9947+/-4.5590 and 10.2003+/-3.9031, respectively, which was lower than that in those without metastasis (12.9433+/-4.7618 and 15.8436+/-5.6917 respectively, P < 0.01 ). The expression of PTEN was correlative with that of nm23 ( Pearson = 0.370, P < 0.05). CONCLUSION The lower expressions of PTEN and nm23 are related to the metastasis of gallbladder cancer.
Female ; Gallbladder Neoplasms ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; secondary ; Lymphatic Metastasis ; Male ; NM23 Nucleoside Diphosphate Kinases ; Nucleoside-Diphosphate Kinase ; biosynthesis ; genetics ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Tumor Suppressor Proteins ; biosynthesis ; genetics

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