Inhibitory Effect Of Thalidomide On OCI-LY10 Cells And Its Mechanism

Background: The most common type of non-Hodgkin's lymphoma is diffuse large B-cell lymphoma(DLBCL)which has varying degrees of malignancy,heterogeneity,and poor prognosis.At the present,the first-line treatment of DLBCL is CHOP(cyclophosphamide,doxorubicin,vincristine,prednisone)combined with rituximab,but 40% of the patients still have relapsed or drug resistance.How to improve the efficacy and reduce drug resistance is still research hotspots for clinicians.Studies have found that thalidomide has anti-tumor effects and can regulate immunity.In recent years,it has found that thalidomide has anti-tumor potential by inhibiting angiogenesis and promoting apoptosis in vitro and in vivo.Angiogenesis and its dysfunction are markers of cancer and ischemic and inflammatory diseases,which contribute to disease progression[1].Vascular endothelial growth factor(VEGF)is a key factor in angiogenesis.It binds to two kinds of vascular endothelial growth receptors(VEGFR-1 and VEGFR-2)to promote angiogenesis.There are many targeted therapies toward VEGF in the clinic,which have certain curative effects.The BCL-2 family includes a large number of proteins associated with apoptosis.They are broadly classified into three subgroups,one anti-apoptotic and two pro-apoptotic,based on function and BH domain composition.BCL-2 protein belongs to the anti-apoptotic group and plays a central role in the mechanism of apoptosis2,inhibiting its function can promote apoptosis.Thalidomide has been found to inhibit gastric cancer,prostate cancer,pancreatic cancer and non-small cell lung cancer,and its effect may be related to VEGF(Vascular endothelial growth factor)and BCL-2(B cell lymphoma/leukemia-2)protein[3-5].However,the role of thalidomide in the treatment of DLBCL is not clear.Therefore,it is important to study its therapeutic effect in DLBCL and explore its mechanism for clinical treatment.Objective: By detecting the inhibitory effect of thalidomide on diffuse large B-cell lymphoma(DLBCL)cell line OCI-LY10 and its effect on the expression of VEGF and BCL-2,the article analyzes the mechanism to provide experimental evidence for clinical drug use.Methods: 1.The DLBCL OCI-LY10 cell line was cultured and divided into control group and different concentrations of thalidomide treatment group.The inhibition rate of each group was measured by CCK-8(cell counting kit-8)method.The levels of VEGF and BCL-2 protein in each group were determined by blot method.The effects of thalidomide on proliferation,VEGF and BCL-2 protein levels of OCI-LY10 cells were analyzed by statistical methods.2.Peripheral blood samples from patients with newly diagnosed DLBCL by pathology at the Second Hospital of Dalian Medical University from 2017.12-2018.12.Exclusion criteria: patients with infection or immunodeficiency and immunosuppression.The control group is from health checkups in our hospital.They have no cancer history before,and no infections and drug usage in the past four weeks.Significant difference in age didn't exist between the two groups.Serum VEGF levels were measured by Elisa,and differences between the two groups were analyzed by One-Way ANOVA.Result: 1.Thalidomide has a proliferation inhibitory effect on DLBCL OCI-LY10 cells(the concentration of each group is 100 ug/ml,150 ug/ml,200 ug/ml,250 ug/ml,the inhibition rates at 48 hours are 0.14±0.32,0.25±0.10,0.38±0.04,0.40±0.06 and at 72 hours are 0.07±0.05,0.25±0.38,0.44±0.16,0.67±0.12,respectively,P<0.05),which is concentration-dependent.2.Compared with the control group,the VEGF protein level of the experimental group(thalidomide treatment group)is significantly decreased(P<0.05),and the inhibitory effect is enhanced with the increase of drug concentration.3.There was no significant difference in BCL-2 protein level between the thalidomide treatment group and the control group(P>0.05).4.There was no significant difference in serum VEGF protein levels between patients with DLBCL and healthy subjects(P>0.05).Conclusion: 1.Thalidomide has a proliferation inhibitory effect on DLBCL OCI-LY10 cell line in a concentration-dependent manner,and its mechanism may be related to the inhibition of VEGF expression;2.Thalidomide has no effect on the expression of BCL-2 in OCI-LY10 cells.3.There is no significant difference in the serum VEGF protein levels between the serum of DLBCL patients and healthy subjects in this study.It is necessary to increase the number of patients.