【作者】 张正茂； 张超； 单保恩； 程建新；

【Author】 Zhang Zheng-Mao Zhang Chao Shan Bao-En Cheng Jian-Xin Department of Obsterics and Gynecology, the fourth hospital of Hebei medical university & Hebei provincial tumor hospital, Shijiazhuang, 050011, China

【机构】 河北医科大学第四医院暨河北省肿瘤医院妇产科；

【摘要】 目的:CD40与CD40L间的结合反应可以启动机体的免疫应答,在宿主抗肿瘤免疫反应中起着重要作用。然而, 一些肿瘤细胞,如表达CD40L的卵巢癌细胞,与宿主相互作用的分子机制仍不十分清楚。本研究旨在探讨转染CD40L cDNA的卵巢癌细胞株OVHM(CD40L／OVHM)的生物学特性及抗肿瘤效应。方法:用脂质体转染法将CD40L cDNA 转入卵巢癌细胞株OVHM,MTT法检测CD40L／OVHM和 OVHM细胞的增殖反应,流式细胞技术检测CD40L／OVHM 和OVHM细胞中CD40、CD40L、MHC-I、MHC—II、CD80、 CD86的表达水平。ELISA法测定皮下接种OVHM或CD40L／ OVHM细胞的小鼠脾细胞和经放射线照射的OVHM细胞共培养上清中IFN-γ的释放量。结果:转染和未转染CD40L 的OVHM细胞体外增殖反应性无显著性差异(p>0．05)。转染CD40L cDNA的卵巢癌细胞CD40L／OVHM与亲本 OVHM相比,共刺激分子CD80、CD86的表达明显增强(P <0．05);将接种CD40L／OVHM的B6C3F1小鼠脾细胞和经放射线照射的OVHM细胞体外混合培养,分泌IFN-γ的水平(1802±187．7pg／ml)明显高于接种OVHM细胞和未接种肿瘤细胞的小鼠脾细胞所分泌IFN-γ的量(20．3±7．9pg／ ml、100．3±19．9 pg／ml)。将CD40L／OVHM细胞接种于 B6C3F 1小鼠后,肿瘤的生长速度与接种OVHM细胞的 B6C3F1小鼠相比明显减慢,体积较小,且无腹腔转移结节, 大部分小鼠无肿瘤细胞生长(P<0．05);将CD40L／OVHM 细胞和OVHM细胞接种BALB／c裸鼠,其肿瘤的生长速度、大小无明显差异(P>0．05)。结论:转染CD40L基因后,可提高卵巢癌细胞的抗原性,引起较强的特异性CTL反应。更多还原

【Abstract】 Objective: Evidence is accumulating that indicates the importance of CD40-CD40L receptor-ligand pair in the initiation and regulation of the human immune response. Whether triggering CD40-CD40L signaling pathway in tumorgensis can cause antitumor effects in ovarian cancer is not clear. To Investige the biological and antitumor effects of ovarian cancer cells line transfected with CD40L cDNA. Methods: The CD40L cDNA was transfected into OVHM cells by lipofectamine 2000, cells proliferation was detected by MTT assay. Immunofluo-rescence technique and flow cytometry were used to evaluated the expression of CD40, CD40L, MHC- I, MHC-II, CD80, CD86.The CD40L/OVHM and OVHM cells were inoculated in the C57BL/6N × C3H/He F1 ( B6C3F1) mice and Balb/c nude mice simultaneously and contralaterally, the tumor volumes was measured. The production of IFN- γ which came from the spleen cells respectively from the CD40L/OVHM- and OVHM cells-inoculated mice cultured with CD40L/OVHM and OVHM cells were measured with an enzyme-linked immunosorbent assay. Restults: Expression of CD40L on OVHM/CD40L cells was much higher than that of OVHM cells, Moreover the expression of costimulatory molecules CD80, CD86 on CD40L/OVHM was enhanced and have a favored the antiutumor immune response. In vitro the proliferation rate of parent and CD40L expressed OVHM cells are not different. A delay in tumor growth was observed when CD40L-expressing OVHM cells and OVHM cells were injected simultaneously and contralaterally. But the tumor growth of OVHM and CD40L/OVHM in Balb/c mice was not different. It is also shown that in vivo immunization of immnuocompetent mice ( B6C3F1) with CD40L/OVHM improved the generation of cytotoxic lymphocytes against parent ovarian cancer ceUs. Conclusion: The forced expression of CD40L cDNA in ovarian tumor cells line OVHM can increase its immunogenicity, and produce the antitumor effects against OVHM-incubated mice.更多还原