| ObjectiveThis study focused on the expression level of serum vascular endothelial growth factor(sVEGF) and serum basic fibroblast growth factor(sbFGF) in non Hodgkin lymphoma(NHL) patients and the relationship between them and clinical features; approach the affects of curcumin on proliferation and VEGFs′expression in Raji cells. The study provides a theory evidence for therapy of anti-vessel neogenesis in clinic.MethodsWe measured the sVEGF and sbFGF in NHL patients by ELISA, and collect clinical materials and analyze the data with SPSS. After Raji cells were treated with curcumin at different concentrations(5umol/L,10umol/L,25umol/L,50umol/L) and different time (24h, 48h,72h), cell proliferation was tested by WST-1 assay; the morphological changes of cells were observed by inverted phase contrast microscope; the apoptosis of Raji cells was studied by Annexin V/PI double-labeled flow cytometry; the change of VEGF contents in cultured supernatants of Raji cells treated with different concentratins of curcumin were measured by ELISA.Results1. The sVEGF levels in NHL patients(1) The sVEGF levels in untreated 36 NHL patients is 443.59±203.48pg/ml, significantly higher than that of healthy controls(90.50±21.42pg/ml)(P<0.01).(2) The sVEGF levels in 17 complete remission(CR) patients is 96.34±38.84pg/ml, 11 partial remission(PR) patients is 109.54±46.37pg/ml, and 8 palindromia(PD) patients is 553.96±250.08pg/ml. The sVEGF levels of untreated NHL patients was significantly higher than that of patients in CR (P<0.01), and was also higher than that of patients in PR(P<0.05); the sVEGF levels with PD was significantly higher than that of healthy controls(P<0.01), and was also higher than that of untreated patients, but no significance between them(P=0.526).(3) The sVEGF levels in 9 CR patients in the first cycle chemotherapy is 152.11±57.98 pg/ml, significantly lower than the sVEGF levels in 27 not CR patients in the first cycle chemotherapy (540.75±124.45 pg/ml)( P<0.05).(4) The sVEGF levels in NHL patients with advanced Ann Arbor stage(P<0.05),high IPI scores(P<0.05) were significantly higher. Compared with those with normal LDH levels, sVEGF levels in NHL patients with higher LDH were elevated(P<0.05); Compared with those with normalβ2-Mg levels, sVEGF levels inβ2-Mg patients with higherβ2-Mg were elevated(P<0.05). But there were no difference about sVEGF levels in untreated NHL patients with different gender, age, B symptoms and number of extranodal site involved(P>0.05).2. The sbFGF levels in NHL patients(1) The sbFGF levels in untreated 36 NHL patients is 314.47±132.01pg/ml, significantly higher than that of healthy controls(78.55±36.35pg/ml) (P<0.05).(2) The sbFGF levels in 17 CR patients is 82.28±34.85pg/ml, in 11 PR patients is 126.21±61.02pg/ml, and in 8 PD patients is 338.40±161.12pg/ml. The sbFGF levels of untreated NHL patients was significantly higher than that of patients in CR and in PR(P<0.05); the sbFGF levels with PD was significantly higher than that of healthy controls(P<0.01), and was also higher than that of untreated patients, but no significance between them(P=0.246).(3) The sbFGF levels in 9 CR patients in the first cycle chemotherapy is 151.14±35.09 pg/ml, significantly lower than the sbFGF levels in 27 not CR patients in the first cycle chemotherapy (346.39±121.40pg/ml)( P<0.05).(4) The sbFGF levels in NHL patients with advanced Ann Arbor stage(P<0.05),high IPI scores(P<0.05) were significantly higher. Compared with those with normal LDH levels, sbFGF levels in NHL patients with higher LDH were elevated(P<0.05); Compared with those with normalβ2-Mg levels, sbFGF levels inβ2-Mg patients with higherβ2-Mg were elevated(P<0.05). But there were no difference about sbFGF levels in untreated NHL patients with different gender, age, B symptoms and number of extranodal site involved(P>0.05).3. The correlation between sVEGF and sbFGFThere is positive correlation between sVEGF level and sbFGF level in NHL patients(r=0.548,P<0.05).4. The change of VEGF contents in cultured supernatants of Raji cells The change of VEGF contents in cultured supernatants of Raji cells treated with different concentratins of curcumin(10μmol/L, 25μmol/L) were measured by ELISA, the levels of VEGF were 369.73±14.22pg/ml, 178.24±21.18pg/ml, and significantly lower than that of the control group′s (707.92±55.92pg/ml).5. The results of WST-1There was no obviously refrain proliferation of Raji cells in 5umol/L curcumin; but in certain range of concentration (10μmol/L, 25μmol/L, 50μmol/L ), curcumin could obviously refrain proliferation of Raji cells in a dose and time dependent manner.6. The morphological changes of Raji cellsThe morphological changes of cells were observed by inverted phase contrast microscope, we can found that the Raji cells turned into disorder from conglobation, the cell population obviously was decreased and the cell volume shrinked, the refraction weakened, the cell membrance disrupted, and we can see cell debris.7. The apoptosis of Raji cellsThrough flow cytometry, we can found that the apoptosis rate of Raji cells was increased as the concentration of curcumin and action time were increased.Conclusions1. Serum VEGF,bFGF levels of NHL patients were higher than that of healthy adults, and were associated with advanced Ann Arbor stage, high IPI scores, LDH, andβ2-Mg. Angiogenesis perhaps involved in the development of NHL, and might be used to monitor patients conditions,evaluate prognosis with other indexes.2. Curcumin can inhibit VEGF secretion of Raji cells , its relied on concentrations. These results suggest that curcumin can anti-vessel neogenesis.3. Curcumin can significantly inhibit Raji cells proliferation in certain range of concentration, its relied on concentrations and times. |
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