Pro-oncogenic potential of NM23-H2 in hepatocellular carcinoma

NM23 is a family of structurally and functionally conserved proteins known as nucleoside diphosphate kinases (NDPK). There is abundant mRNA expression of NM23-H1, NM23-H2, or a read through transcript (NM23-LV) in the primary sites of hepatocellular carcinoma (HCC). Although the NM23-H1 protein is implicated as a metastasis suppressor, the role of NM23-H2 appears to be less understood. Thus, the aim of this study was to examine whether NM23-H2 is associated with hepatocarcinogenesis. The level of NM23-H2 expression in tumor tissues and the surrounding matrix appeared to be independent of etiology and tumor differentiation. Its subcellular localization was confined to mainly the cytoplasm and to a lesser extent in the nucleus. Ectopic expression of NM23-H2 in NIH3T3 fibroblasts and HLK3 hepatocytes showed a transformed morphology, enhanced focus formation, and allowed anchorage-independent growth. Finally, NIH3T3 fibroblasts and HLK3 hepatocytes stably expressing NM23-H2 produced tumors in athymic mice and showed c-Myc over-expression. In addition, NF-kappaB and cyclin D1 expression were also increased by NM23-H2. Lentiviral delivery of NM23-H2 shRNA inhibited tumor growth of xenotransplanted tumors produced from HLK3 cells stably expressing NM23-H2. Collectively, these results indicate that NM23-H2 may be pro-oncogenic in hepatocarcinogenesis.

Clinical application of serum GFAP,NDKA and PARK7 in patients with ischemic stroke / 中华检验医学杂志

Objective To explore the relationship of GFAP, NDKA and PARK7 serum concentrations of patients with IS, and their diagnose and prognosis value in IS. MethodsThe serum concentrations of GFAP, NDKA and PARK7 were detected in 37 IS patients, 28 ICH patients and and 30 healthy persons by ELISA. These indexes of patients were detected in 12 hours, 3 d and 14th day after onset of ischemic stroke. Their neurological injury status were also evaluated by MESSS at corresponding time points, and their activities of daily living were evaluated by BI at 14 d discharge from hospitaL At the same time, the diagnostic efficiency was analysed for IS using the three biomarkers and the combined detection. ResultsIn IS group, the serum concentrations of GFAP in 12 hours, 3rd and 14th day after onset were (5. 49 ±2. 25 )μg,/L, (5. 17 ± 2. 29) μg/L and (5. 96 ± 2.39 ) μg/L, respectively. The serum concentrations of NDKA were 9. 15(6.28 -12.79) μg/L, 9. 13(6.31 - 12.23) μg/L, 9.31(6.40 - 11.83) μg/L respectively,and the serum concentrations of PARK7 were (32. 71 ±6. 34 ) μg/L, (31.23 ±6. 04) μg/L, (32. 79 ±6. 94) μg/L respectively. The serum levels of GFAP, NDKA and PARK were respectively (4. 62 ± 1. 56)μg/L, 4. 24(3. 30 -5. 61 ) μg/L, ( 14. 25 +2. 65) μg/L in healthy control group. The levels in IS groups were remarkably increased compared with the healthy control group except the level of GFAP in the 3rd day (t = 1. 129, P＞0. 05). The levels in other time points were significantly different between patients group and healthy control. t value of GFAP were respectively 2. 642, 1. 870,P＜0. 05; Z value of NDKA were 6. 173, 6.100, 6.278,P ＜0. 01; t value of PARK7 were 14.964, 15.367,16.060, P ＜0. 01. The specificity and sensitivity of the individual detection for diagnosis of IS was 46. 7％ (14/30) and 81.1％( 30/37 ) for GFAP, 90. 0％ ( 27/30 ) and 78.4％ ( 29/37 ) for NDKA, 96. 7％ (29/30) and 97.3％ ( 36/37 )for PARK7. The specificity and sensitivity for combined detection of 3 biomarkers was 96. 7％ (29/30) and 100％ (37/37). The combined detection achieved better specificity and sensitivity. Moreover, the risk of IS with higher level GFAP was 1. 3 times that of the controls ( OR = 1. 300, P = 0. 044 ). The risk of higher NDKA was 1.7 times higher( OR = 1. 668, P = 0. 036 ). The risk of higher PARK7 was 1.8 times higher (OR = 1. 809, P =0. 005 ). The serum levels of GFAP were significantly different between IS and ICH in 12 h(t= 4.097, P=0.000). The serum concentrations of GFAP, NDKA and PARK7 were positively correlated with MESSS score at different time points. In IS, r value were 0. 534, 0. 482, 0. 357 , P ＜ 0. 05at less than 12 h; r value were 0.433, 0.487, 0. 299,P value were 0. 007, 0. 002, 0.073 at 3 d;r value were 0. 394, 0. 200, 0. 084,P value were 0.016, 0. 236, 0.620 at 14 d. And the serum levels of GFAP,NDKA and PARK7 were negatively correlated with BI score at 14th day, r value were -0. 430, -0. 321,-0.076,P value were 0.044,0.050,0.657. Conclusions The concentrations of GFAP, NDKA and PARK7 in serum are closely related with IS. The increased seruro levels of these indexes are risk factors in IS. The detection of these indexes could be helpful for the early diagnosis, timely treatment and prognosis assessment for IS.

NM23 protein expression in colorectal carcinoma using TMA (tissue microarray): association with metastases and survival / Expressão da proteína NM23 no carcinoma colorretal preparado em TMA (tissue microarray): associação com metástases e sobrevivência

CONTEXT: NM23, a metastasis suppressor gene, may be associated with prognosis in patients with colorectal carcinoma. OBJECTIVE: To analyze NM23 expression and its association with the presence of lymph node and liver metastases and survival in patients operated on for colorectal carcinoma. METHODS: One hundred thirty patients operated on for colorectal carcinoma were investigated. Tissue microarray blocks containing neoplastic tissue and tumor-adjacent non-neoplastic mucosa were obtained and analyzed by immunohistochemical staining using a monoclonal anti-NM23 antibody. Immunohistochemical expression was assessed using a semiquantitative scoring method, counting the percentage of stained cells. The results were compared regarding morphological and histological characteristics of the colorectal carcinoma, presence of lymph node and liver metastases, tumor staging, and patient survival. Statistical analysis was performed using the Mann-Whitney test, the Kruskal-Wallis test and Fisher's exact test. Survival analysis was performed using the Kaplan-Meier method and the log-rank test. RESULTS: NM23 expression was higher in colorectal carcinoma tissue than in adjacent non-neoplastic mucosa (P<0.0001). NM23 protein expression did not correlate with degree of cell differentiation (P = 0.57), vascular invasion (P = 0.85), lymphatic invasion (P = 0.41), perineural infiltration (P = 0.46), staging (P = 0.19), lymph node metastases (P = 0.08), or liver metastases (P = 0.59). Disease-free survival showed significant association (P = 0.01) with the intensity of NM23 protein immunohistochemical expression in colorectal carcinoma tissue, whereas overall survival showed no association with NM23 protein expression (P = 0.13). CONCLUSIONS: NM23 protein expression was higher in neoplastic colorectal carcinoma tissue than in adjacent non-neoplastic mucosa, showing no correlation with morphological aspects, presence of lymph node or liver metastases, colorectal carcinoma staging, or overall survival. Disease-free survival was higher in patients with increased NM23 expression.

CONTEXTO: O NM23, denominado de gene supressor de metástases, pode estar relacionado com o prognóstico em doentes com carcinoma colorretal. OBJETIVOS: Analisar a expressão do marcador tumoral NM23 relacionando-a com a presença de metástases linfonodais e hepáticas e com a sobrevivência dos doentes operados por carcinoma colorretal. MÉTODO: Cento e trinta doentes operados por carcinoma colorretal foram analisados. Blocos de "tissue microarray" foram obtidos com tecido neoplásico e com mucosa não neoplásica adjacente ao tumor e submetidos ao estudo imunoistoquímico com o anticorpo monoclonal NM23. A imunoexpressão foi avaliada por método semiquantitativo, com contagem do percentual de células coradas. Os resultados encontrados foram relacionados com as características morfológicas e histopatológicas do carcinoma colorretal, presença de metástases linfonodais e hepáticas, estádio e sobrevivência dos doentes. O estudo estatístico foi realizado com os testes de Mann-Whitney, Kruskal-Wallis e exato de Fisher. A análise da sobrevivência foi calculada pelo método de Kaplan-Meier e pelo teste de long-rank. RESULTADOS: A expressão do marcador NM23 foi maior no tecido do carcinoma colorretal do que na mucosa não-neoplásica adjacente (P<0,0001). A expressão da proteína NM23 não apresentou relação com o grau de diferenciação celular (P = 0,57), invasão vascular (P = 0,85), invasão linfática (P = 0,41), infiltração perineural (P = 0,46), estádio (P = 0,19), metástases linfonodais (P = 0,08) ou metástases hepáticas (P = 0,59). A sobrevivência livre de doença mostrou relação significante (P = 0,01) com a intensidade de imunoexpressão da proteína NM23 no tecido do carcinoma colorretal, e a sobrevivência global não mostrou relação com a expressão da proteína NM23 (P = 0,13). CONCLUSÕES: A expressão da proteína NM23 foi mais intensa no tecido neoplásico do carcinoma colorretal do que na mucosa não-neoplásica adjacente. A expressão da proteína NM23 não se relacionou com os aspectos morfológicos, presença de metástases linfonodais ou hepáticas, estádio do carcinoma colorretal ou com a sobrevivência global. A sobrevivência livre de doença foi maior nos doentes com expressão aumentada do gene supressor de metástases NM23.

The application of proteomic technology in differentiation and early diagnosis of myelocytic leukemia / 中华检验医学杂志

Objective To figure out the differentially expressed proteins using proteome technology in leukemia cells induced into different lineages and investigate the application value in early screening of leukemia.Methods With induction of ATRA and NSC67657, the differentiation models was constructed using HL60 cells which has the potentiality to be induced into different lineages by different inducers.Then the differentially expressed proteins in the process of differentiation was separated using two-dimensional electrophoresis and identified using MALDI-TOF MS.The expression of 3 proteins FE1A1, TLE1, NME3 were chosen to be verified in myeloid samples of 5 leukemia patients and 1 normal volunteer using RT-PCR and WB.Results WB showed that NME3 was differentially expressed after both granulocytic and monocytic differentiation( Normal A value = 0.227, NSC67657 A value= 0.079, ATRA A value = 0.064, P ＜ 0.01 ).However, only in 4 of 5 tested patients' myeloid samples, the NME3 protein expression were differentially expressed compared to the normal myeloid sample( Normal A value = 0.082,2 acute leumia transferred from chronic granulocytic leumia A value = 0.274,0.269, acute monocytic leukemia A value = 0.297, one patient with chronic granulocytic leukemia A value = 0.258.There was significant difference between normal and leukemia group, P ＜0.05 ).A value was 0.121 for another patents with chronic granulocytic leukemia The NME3 protein expression was not differentially expressed compared to the normal myeloid sample,P ＞0.05.Conclusions It is still a long way to go for proteome technology from basic research to clinical application.However, the identification of NME3 protein related to differentiation in leukemia patients' myeloid samples had set the foundation for the early diagnosis of leukemia using proteome technology.