| Leukemia is a heterogeneous hematopoietic malignancy resulting from the blocked differentiation and apoptosis of hematopoietic stem cells and progenitor cells, and malignant cloning hyperplasia. At present, the morbidity of leukemia in China is 3-4 per 100,000 population. Regarding the different mortality rates for malignant neoplasms, leukemia ranks fifth, sixth, and first among males, females, and those aged≤35 years, respectively. The research shows that, as in solid tumors, the occurrence and development of neoplastic hematologic disorders is closely related to angiogenesis and signal transduction. At present, scholars are gradually turning their attention to this area of research. During neoplasia, abnormal angiogenesis plays a vital role. Tumor growth depends on angiogenesis. Tumor cells have the potential to induce angiogenesis. Such potential can only be activated by the stimulation of a growth factor. Basic fibroblast growth factor (bFGF; also known as bFGF-2) is one of the strongest angiogenic factors, and is quite an important positive regulatory factor. In recent years, some research has indicated that the bFGF level in the body fluid of leukemia patients is obviously higher than that of the control group, and that the bone marrow shows obvious angiogenesis. Various in vivo and in vitro experiments have revealed that bFGF accelerates the proliferation and activity of vascular endothelial cells. Researchers have found that leukemia cells exhibit abnormal expression of bFGF and/or bFGF receptor (bFGFR). bFGF can directly stimulate leukemia cells through autocrine or paracrine mechanisms. Further, it can accelerate the proliferation of tumor cells by strengthening the signal transduction level of tyrosine kinase. The new vessels provide continuous nourishment and eliminate any metabolites for the tumor. Thus, bFGF plays an important role in the occurrence, metastasis and prognosis of the tumor. Vascular proliferation is an important pathophysiological process. Vascular proliferation of the bone marrow is related to a poor prognosis in AML, ALL and CML. In recent years, research into the relationship between the vascular proliferation of the bone marrow and vascular endothelial growth factor (VEGF) in patients with AML and CML has revealed that, compared with the control group, the micro vessel density of the bone marrow increases significantly in different leukemia patients, and that it has an obvious relationship with VEGF. Of these patients, the microvessel density and VEGF level of the patients with CML were found to increase significantly. Cyclooxygenase-2 (COX-2) can accelerate the occurrence and development of the tumor, and can lead to an increase in its expression in the leukemia. COX-2 has a tentative association with VEGF expression and vascular proliferation in leukemia—an area of research that needs further exploration and verification. COX-2 induces arachidonic acid to produce its main metabolite, PGE2, which plays a role in the angiogenesis of tumors and induces angiogenesis, such as VEGF and bFGF synthesis. The proliferation, differentiation, maturation, and apoptosis of the hemocyte are controlled by a complex system involving positive or negative cytokines that exert a stimulatory or inhibitory effect, respectively. At present, transforming growth factor-β1 (TGF-β1) is known to exert the strongest negative effect on hemocyte proliferation. Many kinds of normal hemocytes (e.g. monocytes, lymphocytes, blood platelets) can generate TGF-β1, which can not only stimulate growth of the fibroblast and repair of the trauma, but can also influence hematopoiesis and inhibit the proliferation of tumor cells.Currently, most studies focus on a single factor, or two or three factors, among COX-2, bFGF, TGF-β1 and VEGF, and discuss the clinical significance. However, new theories are scarce and of limited scope. This aims of this study were to:examine the change in the COX-2, bFGF, TGF-β1 and VEGF gene expression of the leukemia cell, and the level of COX-2, bFGF, TGFβ1 and VEGF in the serum of leukemia patients; study the relative importance of the four factors; analyze the significance of each test index in the clinical treatment of leukemia; and discuss the relationship between the occurrence/development of leukemia and angiogenesis.Detection and clinical significance of serum cyclooxygenase2、basic fibroblast growth factor、transforming growth factor-β1and vascular endothelial growth factor in acute leukemiaObjective To measure the serum levels of COX-2,bFGF,TGF-β1 and VEGF in patients at different stages of acute leukemia, and discuss the diagnosis, treatment and prognosis of the disease.Method The serum levels of COX-2、bFGF、TGF-β1 and VEGF in 90 acute leukemia patients and 40 healthy adults were determined by ELISA, and the results were analyzed at different stages of the disease. The nuclear cells were counted in a bone marrow smear of 90 patients with acute leukemia.ResultsCompared with the normal control group, the serum levels of COX-2 were found to be elevated before treatment in the acute leukemia patients(P<0.001, t/χ2=6.082). Also, compared with the normal control group, the plasma levels of TGF-β1 were found to be decreased before treatment in the acute leukemia patients (P<0.001, t/χ2=860.000). Moreover, the bFGF and VEGF levels were found to be higher in the acute leukemia group than in the normal control group (P<0.001, t/x2=820.000; P<0.001, t/χ2=4133.000). However, no differences were found in the levels of COX-2、bFGF、VEGF and TGF-β1 between the AML group and the ALL group (P=0.586,χ2=1077.000; P=0.646,χ2= 2906.500; P=0.986,χ2= 1135.500; P=0.729 χ2=2919.000). Also, compared with the normal control group, no differences were found in the levels of COX-2, bFGF、VEGF and TGF-β1 in the patients with acute leukemia after complete remission (CR) (P=0.806, χ2=3756.000;P=0.868,χ2= 2173.500; P=0.905,χ2= 2181.000; P= 0.895, χ2= 3774.000), yet before treatment, differences were found between the levels of COX-2, bFGF、VEGF and TGF-β1 in the patients with acute leukemia after complete remission (CR) (P<0.001, F=61.802; P<0.001, F=1342.108; P<0.001, F= 458.935; P<0.001, F=1349.146). There was a positive correlation between the total number of primary and naive cells and the serum levels of COX-2、bFGF and VEGF in the patients with acute leukemia (R=0. 0.303, P=0.005;R=0.435, P<0.001;R=0.293, P=0.006). In addition, a negative correlation was found between the total number of primary and naive cells and the serum level of TGF-β1 in these patients (R=-0.379,P<0.001). Moreover, a negative correlation was found between the serum levels of COX-2、bFGF, VEGF and TGF-β1 in the patients with acute leukemia (R=-0.401, P<0.001; R=-0.578, P=0.001;R=-0.388, P<0.001). All data were analyzed using SPSS20.0 statistical analysis software.ConclusionOur study showed the serum levels of angiogenesis-promoting factors COX-2, VEGF and bFGF to be elevated before treatment in the acute leukemia patients, but after complete remission, the serum levels of these angiogenic factors were found to be significantly decreased. Further, the serum levels of TGF-β1 were found to decrease before treatment in the acute leukemia patients, but after complete remission, the serum levels were found to be significantly elevated. Angiogenesis may play a role in the leukemogenic process. Therefore, monitoring the levels of serum COX-2、bFGF、TGF-β1 and VEGF at different stages of acute leukemia can provide clinical evidence of disease progression.Detection and clinical significance of serum cyclooxygenase2、basic fibroblast growth factor、transforming growth factor-β1and vascular endothelial growth factor in chronic myeloid leukemiaObjective To measure the serum levels of COX-2、bFGF、TGF-β1 and VEGF in patients at different stages of chronic myeloid leukemia, and discuss the diagnosis、treatment and prognosis of the disease.Method The serum levels of COX-2、bFGF、TGF-β1 and VEGF in 50 chronic myeloid leukemia patients and 40 healthy adults were determined by ELISA, and the results were analyzed at different stages of the disease.ResultsCompared with the normal control group, the plasma levels of COX-2 were found to be elevated before treatment in the chronic myeloid patients (P <0.001,t/χ2=34.862). Also, compared with the normal control group, plasma levels of TGF-β1 were found to be decreased before treatment in the chronic myeloid leukemia patients (P<0.001, t/χ2=1289.000). Moreover, the bFGF and VEGF levels were found to be higher in the acute leukemia group than in the normal control group (P<0.001, t/χ2=820.000; P<0.001, t/χ2=1402.000). Yet when the chronic phase group was compared with the accelerated phase group, differences were found in the levels of COX-2、bFGF、TGF-β1 and VEGF (P<0.001,t/χ2=-6.117; P<0.001, t/χ2= 575.000; P<0.001, t/χ2= 445.000; P<0.001, t/χ2=65.000), Also, when the chronic phase group was compared with the blast crisis group, differences were again found in the levels of COX-2、bFGF、 TGF-β1 and VEGF (P<0.001,t/χ2=-6.974; P<0.001, t/χ2=485.000; P<0.001, t/χ2=465.000; P<0.001, t/χ2=55.000). Compared with the normal control group, no differences were found in the serum levels of COX-2 and bFGF in the patients with chronic myeloid leukemia after complete remission (CR) (P= 0.950, t/χ2=-0.063; P=0.983,t/χ2= 1518.000). Moreover, compared with the normal control group, no differences were found in the levels of TGF-β1 and VEGF in the patients with chronic myeloid leukemia after complete remission (CR) (P= 0.584, t/χ2= 1468.500; P=0.742, t/χ2=1299.000). Compared with the no remission group, differences were found in the serum levels of COX-2、 bFGF、VEGF and TGF-β1 in the patients with chronic myeloid leukemia after complete remission(CR)(P<0.001, t/χ2=-28.355; P<0.001, t/χ2= 630.000; P<0.001, t/χ2= 137.000; P>0.001, t/χ2= 123.000). Finally, a negative correlation was found in the serum levels of COX-2、bFGF and VEGF and TGF-β1 in the patients with chronic myeloid leukemia (R=-0.315, P=0.026;R=-0.330, P=0.020; R=-0.632, P<0.001).All data were analyzed using SPSS20.0 statistical analysis software.ConclusionOur study showed the serum levels of angiogenesis-promoting factors COX-2, VEGF and bFGF to be elevated before treatment in acute leukemia patients, but after complete remission, the serum levels of these angiogenic factors were found to be significantly decreased. Further, the serum levels of TGF-β1 were found to decrease before treatment in the acute leukemia patients, but after complete remission the serum levels were found to be significantly elevated. Angiogenesis may play a role in the leukemogenic process. Therefore, monitoring the levels of serum COX-2、 bFGF、TGF-β1 and VEGF at different stages of acute leukemia can provide clinical evidence of disease progression.Effect in gene expression of COX、bFGF、TGFβ 1and VEGF of resveratrol on k562 cells of myeloid leukemiaObjective To investigate the mechanism influencing the growth of leukemic cells.Methods Different concentrations of resveratrol and imatinib treated K562 cells were examined by MTT assay, Trypan blue staining, Hoechst 33258 staining and flow cytometry apoptosis. The K562 cells were divided into a control group, an IM monotherapy group (0.1μmol/L,0.2μmol/L), a resveratrol (Res) monotherapy group (50μmol/L, 100μmol/L,200μmol/L) and an IM Joint Res group (IM O.1μmol/L+Res 50μmol/L; IM 0.1μmol/L+Res 100μmol/L; IM 0.1μmol/L+ Res 200μmol/L; IM 0.2μmol/L+Res 50μmol/L; IM 0.2μmol/L+Res 100μmol/ L; IM 0.2μmol/L+Res 200μmol/L). Fluorescence quantitative PCR and Wlstern blot analysis were used to determine the levels of COX-2, and the change in bFGF, TGFβ1 and VEGF, after administration of the resveratrol and imatinib.Results1. The results showed that the concentrations of the toxic combination of K562 cells were higher than the resveratrol-alone group and the imatinib-alone group, and that the mortality rate was highest when the K562 cells were treated with resveratrol +0.2μmol/L cisplatin with 200μmol/mL. With increasing time, treatment with resveratro1 +imatinib significantly increased the rate of cell mortality; the cell death rate was highest at 72 h after treatment.2. The results showed that the cytotoxic effect of K562 cells in combination groups of any concentrations was greater than the effect of both the resveratrol-alone group and imatinib-alone group. The highest mortality rate for the K562 cells was observed in the group comprising 200μmol/mL resveratrol+0.2μmol/L imatinib. However, the rate of K562 cell mortality depended on the period of time since treatment with resveratrol+imatinib. The mortality rate was highest at 72 h after dosing.3. The use of imatinib for CML cell lines resveratrol and caspase-3 were detected, along with resveratrol or imatinib concentration, which caspase-3 activity increased.4. Flow cytometry showed that resveratrol alone, and combined with imatinib, had an apoptotic effect on the human leukemia K562 cells, and with increasing concentrations of resveratrol, the mortality rate of the K562 cells gradually increased. Increasing imatinib concentrations also led to a gradual increase in the mortality rate of Nepal K562 cells.5. The Hoechst 33258 staining assay showed that resveratrol alone, and combined with imatinib, had an apoptotic effect on human leukemia K562 cells, and with increasing concentrations of resveratrol, the mortality rate of the K562 cells gradually increased. Increasing imatinib concentrations also led to a gradual increase in the mortality rate of Nepal K562 cells.6. PCR and Western blot analysis showed that, compared with the control group, the level of COX-2, and the expression of bFGF and VEGF, in the 0.2μmol/L imatinib group decreased significantly, while the level of TGF-β1 mRNA expression significantly increased. The difference was statistically significant (P<0.05). However, with 200μmol/mL resveratrol+0.2μmol/L imatinib acting on the K562 cells, the expression of TGF-pi mRNA expression was significantly increased, while the expression of COX-2, bFGF and VEGF was significantly reduced, and a statistically significant difference was observed when the control group was compared with the 0.2μmol/L imatinib group (P<0.05). All data were analyzed using SPSS20.0 statistical analysis software.Conclusion:The resveratrol and imatinib treated leukemia K652 cell line promotes apoptosis, which may be associated with a significantly increased apoptotic rate.The resveratrol, which inhibits angiogenesis, increases the expression of TGF-β1 mRNA and protein, reduces the expression of COX-2, bFGF, VEGF mRNA and protein, and then accelerates the apoptotic rate of the K562 cells, which indirectly reflects the role of angiogenesis in leukemia. |
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