The Study Of Effects On Melanoma Growth And Immune Function In B16F10 Melanoma Mice

Objective:Eosinophils originate from bone marrow stem cells and differentiate in bone marrow.Eosinophils have the abilitities of anti-parasitic and participation in hypersensitivity.But in recent years,some research reported eosinophilia has been observed in a variety of human cancers,eosinophils of infiltration in tumor tissue have anti-tumor effect.To explore the relationship between eosinophils and the tumor,we developed an experimental model consisting of murine B16 melanoma mice that were injected subcutaneous eotaxin,a potent chemoattractant and activator of eosinophils.Methoods:Melanoma mouse model was established by BALB/c mice,thirty mice was divided into three groups:eosinophilia group;model control group;blank control group.The eosinophil group mice was injected eotaxin (100ng/ml) in the inoculation melanoma cells site every other day(a total of 7 times);the model group mice was injected 0.9%NaCL 0.1ml in the inoculation melanoma cells site(a total of 7 times).In the 16th days the mice were killed:(1) All tumor tissues were processed for routine histological studies using hematoxylin and eosin(H&E)-stained sections of the tissues;tumor cell ultrastructure changes were observed by electron microscopy.(2) MTT assay was used to exminate T,B lymphocyte proliferation function and NK cell function in melanoma mice.(3) TNF-αand IL-10 were detected in serum and tumor tissue of melanoma mice with RIA.Results:1.Compared with model control group(tumor average weigh 1.152±0.086g), melanoma cell grew slower in eosinophil group(tumor average weigh 1.122±0.063g),eosinophil group and model control group tumor size have no significant differences(P＞0.05).2.Compared with blank control group,T,B lymphocyte proliferation of melanoma mice were enhanced significantly in eosinophilia group(P＜0.05).Compared with blank control group,NK cell cytotocity rate of melanoma mice in eosinophilia group is 32.7%,eosinophilia group and blank control group have significant differences(P＜0.05);eosinophilia group and model control group have no significant differences.3.HE staining:microscopic evaluation of the eosinophilia group tumors revealed substantially greater eosinophilia at the interface between tumor and connective tissue than in model control group and patchy necrotic tissue was observed in eosinophilia group tumor.4.Electron microscope results showed that eosinophilia group and model control group melanoma cell have different degrees of apoptosis,tumor cell mitochondria cristae is blurred or disappeared,ultrastructure of tumor cells have no obvious change.5.RIA results showed levels of TNF-αand IL-10 is higher in eosinophilia group(TNF-α:9.887±0.285ng/ml;IL-10:89.7±4.62ng/ml)tumor tissue homogenate than in model group.But levels of TNF-αand IL-10 have no significant differences in serum of the two groups.Conclusions:1.Eosinophils have no significant inhibition action on melanoma cell growth in melanom mice.2 Eosinophils on T,B lymphocyte proliferation and NK cell activity in melanoma mice have no significant effects.