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 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 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 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

OBJECTIVEThe purpose of this study was to establish a stable, high-efficient and low-toxic way of transfecting nm23-H1 into Tca8113 line cells, and then to find out whether nm23-H1 could affect the invasion and metastases ability of Tca8113 line cells.

METHODSnm23-H1 was transfected into Tca8113 line cells with Lipofect. The different expressions of nm23-H1 between transfected and non-transfected line cells were detected by the method of immunohistochemistry. The difference of the invasion and metastases ability between transfected and non-transfected line cells was detected by transwell-room and wash techniques. The change of chemo-sensitivity was evaluated by MTT.

RESULTSUsing pCMV-NEO-BAM system to keep stable expression of nm23-H1, the significant difference of NDPKA expression between transfected and non-transfected Tca8113 line cells was discovered; The metastases ability of transfected Tca8113 line cells decreased significantly; The chemo-sensitivity of transfected Tca8113 line cells to CDDP increased significantly.

CONCLUSIONnm23-H1 can inhibit the metastases of Tca8113 line cells and increase the chemo-sensitivity to CDDP significantly.

Antineoplastic Agents ; pharmacology ; Carcinoma, Squamous Cell ; genetics ; pathology ; Cisplatin ; pharmacology ; Drug Resistance, Neoplasm ; Gene Expression Regulation, Neoplastic ; Humans ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Metastasis ; Nucleoside-Diphosphate Kinase ; Proteins ; genetics ; Tongue Neoplasms ; genetics ; pathology ; Transfection ; Tumor Cells, Cultured

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

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 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