Koppold Bernd, Sauer Georg, Buening Hildegard, Hallek Michael, Kreienberg Rolf, Deissler Helmut, Kurzeder Christian
Department of Obstetrics and Gynecology, University of Ulm Medical School, Frauensteige 14, 89075, Ulm, Germany.
J Cancer Res Clin Oncol. 2006 Dec;132(12):787-94. doi: 10.1007/s00432-006-0127-3. Epub 2006 Jun 23.
Supplementing conventional treatment with gene therapy to induce an immune response might be beneficial to cancer patients. In this study, we evaluated the efficiency of transduction of breast cancer cells with recombinant adeno-associated virus (rAAV) and effects of cytotoxic agents used in chemotherapy. Furthermore, the capacity of tumor cells expressing transgenic CD40 ligand (CD40L) to stimulate dendritic cells was measured.
Breast cancer cell lines were infected with a rAAV encoding the enhanced green fluorescent protein (EGFP) or murine CD40L and transgene expression was analyzed by flow cytometry. Stimulation of isolated human dendritic cells by CD40L-expressing tumor cells was quantified by measuring secreted interleukin 12.
Infection with an EGFP-encoding rAAV resulted in variable fractions (14-93%, mean 42%) of transgene-expressing cells. Pre-incubation of MM 157, MM 231, and MCF7 cells with epirubicin or carboplatin substantially increased AAV-mediated transgene expression. rAAV/CD40L was used to generate CD40L-transgenic tumor cells, which specifically activated immature dendritic cells, as confirmed by blocking with an antibody binding to CD40L.
The efficiency of rAAV-mediated gene transfer into breast cancer cells is significantly higher than previously reported and can be further enhanced by co-administration of chemotherapeutic agents. We also confirmed that breast cancer cells can activate human dendritic cells after infection with a CD40L-encoding rAAV.
用基因疗法补充传统治疗以诱导免疫反应可能对癌症患者有益。在本研究中，我们评估了重组腺相关病毒（rAAV）转导乳腺癌细胞的效率以及化疗中使用的细胞毒性药物的作用。此外，还检测了表达转基因CD40配体（CD40L）的肿瘤细胞刺激树突状细胞的能力。
用编码增强型绿色荧光蛋白（EGFP）或小鼠CD40L的rAAV感染乳腺癌细胞系，并通过流式细胞术分析转基因表达。通过测量分泌的白细胞介素12来量化表达CD40L的肿瘤细胞对分离的人树突状细胞的刺激。
用编码EGFP的rAAV感染导致转基因表达细胞的比例各不相同（14 - 93%，平均42%）。表柔比星或卡铂对MM 157、MM 231和MCF7细胞进行预孵育可显著增加AAV介导的转基因表达。rAAV/CD40L用于产生CD40L转基因肿瘤细胞，经与CD40L结合的抗体阻断证实，该细胞可特异性激活未成熟树突状细胞。
rAAV介导的基因转移到乳腺癌细胞中的效率显著高于先前报道，并且通过联合使用化疗药物可进一步提高。我们还证实，用编码CD40L的rAAV感染后，乳腺癌细胞可激活人树突状细胞。
