PTEN基因不同表达黑色素瘤模型小鼠T淋巴细胞亚群检测及意义

Detection and significance of T lymphocyte subsets in melanoma model mice with different expression of PTEN gene

目的通过检测PTEN基因不同表达黑色素瘤模型小鼠T淋巴细胞亚群的比例及数量变化,探索PTEN在黑色素瘤免疫逃逸中的机制。方法将72只同系C57BL/6小鼠随机分为三组,每组24只:A组注入0.2ml的RPMI-1640培养液;B组注入0.2ml浓度为4×10^6/ml的(PTEN+/+-B16F10);C组注入0.2ml浓度为4×10^6/ml的(PTEN-/--B16F10)。接种后第7日开始每隔5天每组选6只小鼠制备脾脏单细胞悬液,分析CD3^+T、CD4^+T、CD8^+T相对数量及CD28^+/CD8+的比例。结果A组CD3^+T、CD4^+T在D7、D12、D17组内比较无差异,分别与D22比较差异有显著性。CD8^+T、CD28^+/CD8+比例及数量基本平稳。B组、C组各时间点组内比较差异有显著性。B组下降幅度分别为:79.8%、82.6%、65.37%、58.69%。C组下降幅度分别为:89.62%、89.24%、84.85%、70.53%。组间比较D7、D12,C组<B组无差异,D17、D22,C组<B组差异有显著性。结论PTEN基因缺失表达促进了黑色素肿瘤的生长,通过降低小鼠体内T细胞亚群的相对数量,降低了机体的抗肿瘤能力,参与黑色素瘤对机体的免疫逃逸,从而促进了黑色素瘤的发生及发展。

Objective To explore the mechanism of PTEN in the immune escape of melanoma by detecting the proportion and quantity of T-lymphocyte subsets in mice with different expression of PTEN gene.Methods Seventy-two C57BL/6 healthy mice were brought into experiment and divided into three groups,24 mice in each group.Group A injected 0.2ml RPMI-1640 culture medium;group B injected 0.2ml 4×10^6/ml(PTEN+/+-B16F10);group C injected 0.2ml 4×10^6/ml(PTEN-/-B16F10).Six mice were randomly selected every five days since D7,the spleens were removed and cell suspensions were prepared,CD3^+T,CD4^+T,CD8^+T,CD28^+/CD8+value were analyzed.Results There was no significant difference among D7,D12 and D17,and there was significant difference between D22(P<0.05).The proportion of CD8^+T and CD28^+/CD8+were basically stable with time.Compared with group B and C,there was significant difference.In group B,the decrease rate of CD3^+T was 79.8%,CD4^+T was 82.6%,CD8^+T was 65.37%,and CD28^+/CD8+was 58.69%.In group C,the decrease rate of CD3^+T was 89.62%,CD4^+T was 89.24%,CD8^+T was 84.85%,and CD28^+/CD8+was 70.53%.On D7 and D12,group C<group B.On D17 and D22,group C<group B.Conclusion PTEN gene deletion promotes the growth of melanoma.PTEN gene deletion reduced the anti-tumor ability of the body by reducing the relative number of T cell subsets,which participated in the immune escape of melanoma,and thus promoted the occurrence and development of melanoma.