死亡诱骗受体3对乳腺癌预后及乳腺癌细胞的体外功能的影响

Effect of death decoy receptor 3 on prognosis of breast cancer and function of breast cancer cells in vitro

目的观察死亡诱骗受体3(DcR3)对乳腺癌预后的影响和在乳腺癌细胞侵袭功能中的作用。方法通过实时定量聚合酶链反应(PCR)(real-time Q-PCR)进行定量检测包含115例乳腺组织样本的DcR3表达,对应患者随访并分析,中位随访时间10年。通过免疫细胞化学法和RNA提取和逆转录聚合酶链反应(RT-PCR)检测DcR3在乳腺癌MCF7、MDA-MB-231细胞系中的表达,建立DcR3敲除的细胞亚系并验证。通过侵袭和迁移模型观察减少DcR3表达后产生的影响。结果患者分为预后良好组(n=81)和预后不良组(n=26)。预后不良组(133 350±49 646)拷贝/50 ng RNA较预后良好组(5 393±1 428)拷贝/50 ng RNA,P=0.020)DcR3的表达显著增高;病理分级2级的肿瘤较病理分级1级的肿瘤DcR3表达显著升高[(82 844±34 068)拷贝/50 ng RNA(n=39)]与[(5 371±3 500)拷贝/50 ng RNA,n=20,P=0.029]。成功敲除MCF7细胞系和MDA-MB-231细胞系的DcR3基因,DcR3的敲除降低了MCF7细胞的侵袭和迁移能力(P=0.009,P=0.001),但MDA-MB-231细胞系在这两方面的差异无统计学意义(P=0.475,P=0.102)。结论DcR3促进了乳腺癌细胞的侵袭性,在乳腺癌的转移中起到了一定作用,DcR3检测有助于乳腺癌预后判断。

Objective To study the effect of death decoy receptor 3 on the prognosis of breast cancer and the invasive function of breast cancer cells in vitro. Methods Expression of DcR3 were assessed qualitatively by Q-PCR to analyze the correlation in 115 mammary tissue samples with a 10-year median follow-up. The expression of DcR3 was examined in MCF7 and MDA-MB-231 cell lines using immunocytochemical staining and RT-PCR. DcR3 knock-down cell sub-lines were constructed. The effects of reduced DcR3 expression were observed by establishing invasion and migration models. Results Patients were divided into the good prognosis group (n=81) and the poor prognosis group (n=26). The expression of DcR3 in the poor prognosis group (133 350+49 646 copies/50 ng RNA)was significantly higher than that in the good prognosis group (5 393+1 428 copies/50 ng RNA, P=0.020). DcR3 transcripts were found to be increased significantly in grade 2 cancers compared to well differentiated grade 1(82 844±34 068 copies/50 ng RNA, n=39,) vs (5 371±3 500 copies/50 ng RNA, n=20, P=0.029).The DcR3 gene of MCF7 cell line and MDA-MB-231 cell line were successfully knocked out and verified that DcR3 knockout. And the invasion and migration of MCF7 cells were inhibited (P=0.009, P=0.001). However, no significant difference was found in these two aspects of the MDA-MB-231 cell line (P=0.475, P=0.102). Conclusion DcR3 promotes the capacity of invasion of breast cancer cells and plays an important role in the metastasis of breast cancer. DcR3 detection is helpful to the judgment about prognosis of breast cancer.