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负载干细胞抗原的DC联合CIK细胞对乳腺癌荷瘤鼠肿瘤杀伤研究

庞春淼,吕艳,孙雯雯,司玉玲   

  1. 天津市第四中心医院
  • 收稿日期:2013-10-24 修回日期:2014-02-10 出版日期:2014-06-15 发布日期:2014-06-15
  • 通讯作者: 庞春淼

  • Received:2013-10-24 Revised:2014-02-10 Published:2014-06-15 Online:2014-06-15

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摘要： 目的：研究不同乳腺癌细胞抗原负载的DC-CIK细胞对于乳腺癌荷瘤鼠体内肿瘤的杀伤效果及其与Wnt通路的关系。方法：分离乳腺癌细胞系MCF-7/ADR中的干细胞并制备冻融抗原，以此冲击由正常人外周血提取的单个核细胞诱导培养的DC-CIK细胞，注入已建立的MCF-7/ADR乳腺癌荷瘤鼠模型中，并且与普通抗原冲击的DC-CIK细胞，未经抗原冲击DC-CIK细胞，单纯CIK细胞，及生理盐水组建立实验组及对照组，观察此5组中小鼠肿瘤细胞生长情况，通过原位末端标记法（TUNEL）检测各组肿瘤组织凋亡情况，免疫组化法检测bcl-2，Bax及β-catenin表达情况。结果：实验组和对照组小鼠肿瘤体积在治疗后存在差异，同组小鼠肿瘤体积体积在治疗前后也存在差异，对照组小鼠肿瘤体积最大（3.6245±0.09264）cm3，经干细胞抗原冲击的DC-CIK治疗组肿瘤体积最小（1.2342±0.13116） cm3，其tunel染色阳性率最高，bax表达阳性最强，bcl-2，β-catenin表达较其他组最弱。结论：经乳腺癌干细胞抗原冲击的DC-CIK细胞对乳腺癌荷瘤鼠的肿瘤组织较强的杀伤效果，其机制可能是Wnt/β-catenin信号通路中的β-catenin表达降低，并引起bax表达增高，bcl-2表达降低，从而引起肿瘤细胞凋亡。

关键词: 乳腺癌干细胞, DC, CIK, Bcl-2, Bax, β-catenin

Abstract: objective: To investigate the tumor-inhibitory effect and apoptotic pathway of cytokine-induced killer cells (CIK) co-cultured with dendritic cells (DC) loaded breast cancer stem cell antigen on the same tumor-bearing mice. Methods: Separated breast cancer stem cells from a cell line of MCF-7/ADR and then extract freeze-thaw antigen of the stem cell.DC and CIK cells were respectively derived from peripheral blood mononuclear cells (PBMC) of healthy individuals. CIK was co-cultured with DC pulsed or unpulsed by the above antigen lyses.This DC-CIK will injected to tumor-bearing mice which was injected by MCF-7/ADR cell before, and contrast with the common breast cancer antigen pulsed of DC-CIK(SCAP-DC-CIK), without antigen pulsed DC-CIK(AP-DC-CIK), CIK with DC and normal saline (NS).The antitumor effects were evaluated and compared in 5 groups through the tumor size, TUNEL and examining BCL-2, Bax andβ-catenin. Results: The tumor volume of each group was of difference in significance.The tumor volume of treatment group is small than NS group. TUNEL shows that the tumor of SCAP-DC-CIK group positive rate is highest and with the strongest positive bax expression and the weakest BCL-2,β-catenin expression . Conclusion: SCAP-DC-CIK have higher tumor-inhibitory effect than other treatment group.The mechanism may be by reduced expression of β- catenin to effect Wnt/β- catenin signaling pathway, and lead to an increased of bax and reduced of BCL - 2 expression to induce the apoptosis of tumor cell.

Key words: Breast cancer stem cell, DC, CIK, Bcl-2, Bax, β- catenin