The Study Of Expression Of Serum Insulin-like Growth Factor-I And Insulin-like Growth Factor Binding Protein-3and Its Clinical Significance In Non-Hodgkin’s Lymphoma Patient

Background:New diagnosed cases of malignant lymphoma recently increased year by year inChina. The morbility and mortality of lymphoma were ranked the top11-13positionof all malignant tumors, it has become one of common malignant tumors in China.Although we have taken development in the molecular diagnosis of NHL recentyears, there is no golden standard index in the prognosis of lymphoma.Seruminsulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3(IGFBP-3) had showed correlation with many tumors. The incidence of tumor isassociated with cell proliferation, uncontrolled apoptosis and the regulation of cellcycle. Insulin like growth factor axis plays a very important role in the regulation ofcell growth and apoptosis. On the other hand, the method of IGF-IR and otherreceptor-type tyrosine kinase multiple target inhibitors in combination withchemotherapy for patients with tumors became highlight. The research with IGF-Iand IGFBP-3regulation or pathway as a target will provide new strategies andapproaches of prevention and cure for NHL. There were seldom reports about therelationship of IGF-I and IGFBP-3with lymphoma until now. In our study, we would assay serum IGF-I level and serum IGFBP-3level in order to find out thecorrelation with diagnosis, treatment and prognosis of NHL. This study would helpus to know more about the diagnosis, treatment and prognosis of NHL, and providethe basis for targeted therapy of NHL in the future.Objective:We investigated the research of expression and clinical significance of serum IGF-Iand IGFBP-3in NHL. We analysed the result to find out whether it has any relationwith diagnosis, treatment and prognosis of NHL. Our study would help us hnowmore about IGF-I and IGFBP-3regulation or pathway, and provide the basis fortargeted therapy of NHL in clinical applications in the future.Patients and Methods:1. Patients(1)Experimental group: Fifty adult patients with diagnosed NHL between May2009and January2012in Guangzhou First Municipal Hospital and Nanfang Hospital,Southern Medical University were recruited in our study.(2)Control group: Fifty non tumor patients and health examination adults betweenMay2009and January2012in Guangzhou First Municipal Hospital were recruitedas control.2. Serum separation(1) Peripheral blood had been taken3ml from patients in non anticoagulant tubes,centrifuged3000rpm for10minutes to get the upper serum, stored at-25C.(2) Preservation: Preserve blood specimens at-25degrees C below. That are readyfor determining serum IGF-I level and serum IGFBP-3level.3. IGF-I, IGFBP-3detection: Chemical luminescence immunity analysismethod（Double antibody sandwich method ELISA）.(1) IMMULITE1000Automated Immunoassay Analyzer (Diagnostic ProductsCorp, USA) and IMMULITE1000IGF-I kit as well as IMMULITE1000IGFBP-3kit had been utilized to assayed serum IGF-I and IGFBP-3level.（2）The IMMULITE1000system utilized assay-specific murine monoclonal antibody (anti-IGF-I or anti-IGFBP-3McAb) coated plastic beads as the solidphase.The coated bead was housed in a proprietary plastic device,namely a TestUnit.After adding the quantitative tested serum into the Test Unit, incubated thesample with the alkaline phosphatase conjugated the second antibody (polyclonalrabit anti-IGF-I antibody or anti-IGFBP-3McAb) in60minutes or30minutesrespectively, the reaction mixture was separated from the bead by spinning the TestUnit at high sped on its vertical axis.(3) The entire fluid contents (the sample, excess reagent and wash solution) wastransferred to a coaxial waste chamber in the Test Unit. The bead was left with noresidual, unbound label.(4) The amount of bound label was then quantitated with a dioxetane substrate thatproduced light. Put the microporous plate into the analyzer. Through the apparatusinside the three-dimensional transmission system, the photon number of the hole isread from the photon counter in turn.(5) Light emission is measured by a Photomutiplier Tube (PMT) and the results werecalculated for each sample.The molecular concentration to be detected of the samplewill be quantitative analysised according to the standard model.3. Statistics methodsOne-way ANOVA was used for the comparision of more than two groups, and the qtest of the Student-Newman-keuls measurement was taken for the comparision of theaverage of samples. The independent-samples t test was used between the twogroups, and the One-SampleKolmogorov-Smirnov Test had been done. The twosides Spearman test had been taken for the hierarchical grouping data and the twosides Pearson test had been used for the Continuous measurement.Result:一. Comparison with NHLgroup and normal control group(1) Serum IGF-I level:The serum IGF-I level of lymphoma group is109.37±24.87ng/mL. The serum IGF-Ilevel of normal control group is182.86±26.86ng/mL. P<0.0001(Independentsamples T test). T=8.631. The difference has statistics significance. The serum IGF-Ilevel of lymphoma group is lower than normal control group (P<0.01). (2) Serum IGFBP-3level:The serum IGFBP-3level of lymphoma group is2.75±0.72ug/ml. The serumIGFBP-3level of normal control group is4.73±0.52ug/ml. P<0.0001(Independentsamples T test). T=9.002. The difference has statistics significance. The serumIGFBP-3level of lymphoma group is lower than normal control group (P<0.01).(3) IGF-I/IGFBP-3：The ratio of IGF-I/IGFBP-3of lymphoma group is (40.35±4.66)×10-3. The ratio ofIGF-I/IGFBP-3of normal control group is （38.72±4.15）×10-3. P=0.069(Independent samples T test). The difference has not statistics significance (P>0.01).二. Comparison with different groups of NHLpatients.(1) Serum IGF-I level:①The serum IGF-I level of aggressive lymphoma group is108.28±25.98ng/mL. Theserum IGF-I level of indolent lymphoma group is109.17±24.78ng/mL. P＞0.01(Independent samples T test). The difference has not statistics significance.②The difference of serum IGF-I levels of stageⅠ group、stageⅡ group and stageⅢgroup has not statistics significance（p>0.01）.But the serum IGF-I level of stageⅣgroup is lower than other stage groups（p<0.01）.③The difference of serum IGF-I levels of the low risk group、low-intermediate riskgroup and intermediate-high risk group has not statistics significance（p>0.01）.Butthe serum IGF-I level of the high risk group is lower than other risk group（sp<0.01）.(2) Serum IGFBP-3level:①The serum IGFBP-3level of aggressive lymphoma group is2.72±0.74ug/ml. Theserum IGFBP-3level of indolent lymphoma group is2.79±0.78ug/ml. P＞0.01(Independent samples T test). The difference has not statistics significance.②The difference of serum IGFBP-3levels of stageⅠ group、stageⅡ group andstageⅢ group has not statistics significance（p>0.01）.But the serum IGFBP-3level ofstageⅣ group is lower than other stage groups（p<0.01）.③The difference of serum IGFBP-3levels of the low risk group、low-intermediaterisk group and intermediate-high risk group has not statistics significance（p>0.01）.But the serum IGFBP-3level of the high risk group is lower than other risk groups（p<0.01）.(3) The ratio of IGF-I/IGFBP-3：①The ratio of IGF-I/IGFBP-3of aggressive lymphoma group is（40.40±5.59）×10-3.The ratio of IGF-I/IGFBP-3of indolent lymphoma group i（s40.56±4.86）×10-3.P=0.282(Independent samples T test). The difference has not statistics significance(P>0.01).②The difference of the ratios of IGF-I/IGFBP-3of stageⅠ group、stageⅡ group、stageⅢ group and stageⅣ group has not statistics significance（p>0.01）.③The difference of the ratios of IGF-I/IGFBP-3of the low risk group、low-intermediate risk group、intermediate-high risk group and high risk group hasnot statistics significance（p>0.01）.三. Comparison with the Indexs before chemotherapy and the Indexs afterchemotherapy.①Serum IGF-I level before chemotherapy:108.28±25.98ng/ml. Serum IGF-I levelafter chemotherapy:168.97±29.98ng/ml. P <0.0001(The two paired sample T test).The difference has statistics significance.②Serum IGFBP-3level before chemotherapy:2.72±0.74ug/ml. Serum IGFBP-3level after chemotherapy:4.18±0.61ug/ml. P <0.0001(The two paired sample Ttest). The difference has statistics significance.③the ratios of IGF-I/IGFBP-3before chemotherapy:（40.40±5.59）×10-3. the ratiosof IGF-I/IGFBP-3after chemotherapy（:40.52±4.67）×10-3. P=0.856(The two pairedsample T test). The difference has not statistics significance.Conclusion：1. Serum IGF-I level and Serum IGFBP-3level of lymphoma group were significantlylower than normal control group(P<0.01). That May be related to the occurrenceand development of Non-Hodgkin’s lymphoma. That contributes to the diagnosis ofNon-Hodgkin’s lymphoma.2. There was no difference in the ratio of IGF-I/IGFBP-3between lymphoma groupand normal control group (P>0.01). The ratio of IGF-I/IGFBP-3had no obvious correlation with Non-Hodgkin’s lymphoma.3.The difference of serum IGF-I level、Serum IGFBP-3level and the ratio ofIGF-I/IGFBP-3between aggressive lymphoma group and indolent lymphomagroup has not statistics significance.4. Serum IGF-I level and serum IGFBP-3level of stage Ⅳ group and the high riskgroup were significantly lower than other groups. Serum IGF-I level and serumIGFBP-3level reflect the development and evaluation of prognosis ofNon-Hodgkin’s lymphoma in a certain degree.5. Serum IGF-I level and Serum IGFBP-3level were higher after chemotherapy.Serum IGF-I level and Serum IGFBP-3level reflect the efficacy in a certain degree.That contributes to the evaluation of treatment efficacy and prognosis ofNon-Hodgkin’s lymphoma.6. There was no significant difference in the ratio of IGF-I/IGFBP-3between the twogroups(before chemotherapy、after chemotherapy). The ratio of IGF-I/IGFBP-3had no obvious correlation with treatment efficacy of Non-Hodgkin’s lymphoma.