转染CD40配体的卵巢癌细胞株OVHM抗卵巢癌肝转移机制的研究  

Inhibitory effects of enhanced expression of CD40L in ovarian cancer OVHM cells on the liver metastases in mice

作　　者：张正茂[1] 刘洁[1] 张风华[3] 顾来梅 张超[2] 陈书诚[1] 康山 单保恩[2] 

机构地区：[1]河北医科大学第四医院妇产科,石家庄050011 [2]河北医科大学第四医院科研中心,石家庄050011 [3]河北省人民医院普外科

出　　处：《中华肿瘤杂志》2009年第7期505-509,共5页Chinese Journal of Oncology

基　　金：河北省自然科学基金（C2007001058）

摘　　要：目的探讨转染CD40配体（CD40L）的卵巢癌细胞株OVHM（CIMOL—OVUM）体内抗卵巢癌肝转移的可能机制。方法6～8周龄的C57BL／6N×C3H／He杂交一代（B6C3F1）雌性小鼠5只，经小鼠脾脏内接种OVHM，应用HE染色法验证小鼠肝脾转移模型是否建立成功。将OVHM细胞、空载体DNA—pMKITneo—OVHM细胞和CD40L—OVHM细胞接种于B6C3F1小鼠的脾脏，应用流式细胞术，分析荷瘤小鼠脾细胞CD11C分子的表达情况；应用四甲基偶氮唑蓝（MTT）法，检测荷瘤小鼠脾脏细胞毒性T淋巴细胞（CTL）的杀伤活性；应用酶联免疫吸附试验（ELISA），检测荷瘤小鼠外周血血清中干扰素（IFN）-γ、肿瘤坏死因子（TNF）-α、白细胞介素（IL）-12、IL-4和IL—10的含量，并且观察小鼠脾脏和肝脏的成瘤情况及小鼠生存时间。结果经病理学证实，卵巢癌肝脾转移动物模型建立成功。CD40L—OVHM组中小鼠脾细胞CD11C阳性细胞率明显高于OVHM组和转染空载体DNA—pMKITneo—OVHM组。CD40L—OVHM组小鼠脾脏内CTL的特异性杀伤活性明显增强，小鼠外周血血清中IFN-γ、TNF-α和IL-12的含量明显升高，而IL-4和IL-10的含量则明显降低，与OVHM组和转染空载体DNA—pMKITneo—OVHM组相比，差异均有统计学意义（P〈0．05）。CIMOL—OVHM组小鼠的肝脏和脾脏重量均明显低于OVHM组和转染空载体DNA-pMKITneo—OVHM组，并且小鼠的生存期也明显延长（P〈0．05）。结论转染CD40L cDNA的卵巢癌细胞可促进脾脏树突状细胞（DC）的成熟和分化，增强脾脏CTL的特异性杀伤活性，诱导Th1型细胞因子的分泌，抑制Th2型细胞因子的分泌，这可能是CIMOL基因体内抗卵巢癌肝转移的机制之一。Objective To examine the in vivo anti-metastatic effect of enhanced expression of CD40L cDNA in murine ovarian cancer OVHM cells （ CD40L-OVHM ） injected into the spleen on liver metastasis in mice. Methods OVHM cells were inoculated into the spleen of 6 N 8 week-old female B6C3F1 （C57BL/6N x C3H/He） mice. The established liver metastasis was identified by histopathology （ HE staining）. OVHM cells, DNA-pMKITneo-OVHM cells or CD40L-OVHM cells were inoculated into the spleen of female B6C3F1 mice and the expressions of CDllc in splenic cells were detected by flow cytometry. The specific cytotoxicity of splenic cells was detected by MTT assay, and the serum cytokines of IFN-γ, TNF-α, IL-12, IL-4 and IL-10 of the mice were measured by enzyme linked immunoabsorbent assay. The liver metastases and the survival time of the mice were also recorded. Results The mouse models with liver metastasis by injecting tumor cells into the spleen of mice were established. The expression of CD11 c and the specific killing rate in CD40L-OVHM cells group was significantly higher than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group. The expressions of IFN-γ, TNF-α and IL-12 in the CD40L-OVHM cells group were much more increased than OVHM cells group and DNA-pMKITneo- OVHM cells group, but the expressions of IL-4 and IL-10 in the CD40L-OVHM cells group were decreased significantly （P 〈 0.05 ）. The average weights of livers and spleens of mice in CD40L-OVHM cells group were significantly lower than those of DNA-pMKITneo-OVHM cells group and OVHM cells group. The survival time of mice in CD40L-OVHM cells group was also significantly longer than that in the OVHM cells group and DNA-pMKITneo-OVHM cells group （P 〈 0.05）. Conclusion The data directly demonstrate that the expression of CD40L in ovarian cancer cells （CIMOL-OVHM） can enhance the proliferation and differentiation of dendritic cells in the spleen and induce specific cytotoxic effect of T cells in the spleen, and may regulate the immune

关 键 词：CD40配体 小鼠 转基因 卵巢肿瘤 受体 细胞因子 

分 类 号：R686[医药卫生—骨科学]