【摘要】 目的 探讨不同死亡形式的卵巢癌细胞对巨噬细胞协同刺激分子CD80和CD86表达的影响及其可能机制。方法 以紫外线诱导卵巢癌细胞SKOV3凋亡，反复冻融诱导SKOV3细胞坏死。以薄膜-超声法制备含磷脂酰丝氨酸(PS)脂质体及磷脂酰胆碱(PC)脂质体。分别与脂多糖(LPS)作用下的人外周血单核细胞来源的巨噬细胞共同培养后，荧光显微镜下检测巨噬细胞对死亡细胞的吞噬及细胞免疫化学方法检测巨噬细胞CD80和CD86的表达。结果 巨噬细胞对凋亡与坏死的卵巢癌细胞SKOV3都能积极的吞噬，吞噬的比率分别为(35.3±6.9)%和 (31.2±4.1)%。但坏死卵巢癌细胞能显著增强巨噬细胞CD86的表达(P=0.013)，而凋亡卵巢癌细胞却不能增强巨噬细胞CD86表达(P=0.141)。坏死和凋亡卵巢癌细胞对CD80的表达均无显著影响(P=0.377,P=0.914)。PS及PC脂质体对巨噬细胞CD80(P=0.619;P=0.923)和CD86(P=0.414;P=0.816)的表达均无明显影响。结论 不同死亡形式的卵巢癌细胞对巨噬细胞协同刺激分子表达的影响不同，坏死的卵巢癌细胞上调巨噬细胞CD86表达，而凋亡的卵巢癌细胞无此作用。死亡细胞对巨噬细胞CD86表达的影响可能通过非PS途径起作用。

    【关键词】 卵巢癌；细胞死亡；巨噬细胞；协同刺激分子；磷脂酰丝氨酸

    Apoptotic or necrotic of ovarian cancer cell on macrophages CD80 and CD86 expression

    CHEN Xiao-jun,LOU Hai-ya,ZHOU Cai-yun.

    Department of Oncology,Gynaecology and Obstetrics Hospital Affiliated to Zhejiang University Medical College,Hangzhou 10006,China

    【Abstract】 Objective To investigate the differential effect of apoptotic versus necrotic ovarian cancer cells on macrophages CD80 and CD86 expression and its possible mechanisms.Methods The ovarian cancer cell line SKOV3 was induced to apoptosis by ultraviolet rays and necrosis by rapidly freezing-thawing twice.Phosphatidylserine(PS)-liposomes and hosphatidylcholine(PC)-liposomes were prepared by vortexing and sonication.The two types of dead cells were respectively cultured with human monocyte-derived macrophages(HMDM)and LPS.After 24 hours,the phagocytosis assay was performed under fluorescence microscope.The two types of dead cells and two types of liposomes were respectively cultured with HMDM and LPS.After 18～20 hours,the expression of CD80 and CD86 were detected by immunocytochemistry.Results Both apoptotic and necrotic SKOV3 cells could be abundantly phagocyted by macrophages activated by LPS.The percent macrophages positive for uptake of apoptotic or necrotic SKOV3 cells was(35.3±6.9)%,(31.2±4.1)% respectively.However,the necrotic SKOV3 cells,but not apoptotic SKOV3 cells,upregulated the expression of CD86 molecule on the macrophages significantly.Both dead cells did not change CD80 expression on macrophages.The PS-liposome and PC-liposome had no significant effect CD80 and CD86 expression on macrophages elicited by LPS.Conclusion Macrophages activated by LPS can phagocyte apoptotic or necrotic SKOV3 cells,but the two types of dead SKOV3 cells have different effects on macrophages co-stimulating molecules xpression.Phosphatidylserine may not participate in regulation of CD80 and CD86 expression on macrophages by dead tumor cells. ......