紫草素诱导卵巢癌细胞表达钙网织蛋白促进 DC 成熟的研究

[摘要]目的探讨经紫草素诱导凋亡的卵巢癌细胞促进树突状细胞（DC）成熟的作用及其机制。方法分别用不同浓度紫草素以及不舍紫草素的培养基处理卵巢癌HO-8910细胞系。48h后检测各组HO-8910细胞的凋亡率和膜表面钙网织蛋白的表达（流式细胞技术）；同时将各组HO-8910细胞，在加入和未加入钙网织蛋白抑制性结合肽的条件下分别与人外周血来源的树突状细胞混合培养（未经过紫草素处理的H0—8910细胞与树突状细胞的混合培养设为对照组），48h后检测各组与树突状细胞成熟相关的膜分子CD86的表达情况（流式细胞技术）。结果与未经药物处理的HO-8910细胞相比，经紫草素处理的HO-8910细胞凋亡率和膜表面钙网织蛋白的表达显著增加，并且凋亡率和钙网织蛋白水平呈紫草素浓度依赖性。经与紫草素处理过的HO-8910细胞混合培养后，树突状细胞表达CD86较对照组显著上调，同时钙网织蛋白抑制性结合肽能显著抑制CD86的上调。结论经紫草素诱导凋亡的卵巢癌HO-8910细胞能促进树突状细胞的成熟，其机制与诱导凋亡细胞膜表面暴露钙网织蛋白有关。提示在卵巢癌的化疗中使用紫草素，能加强凋亡肿瘤细胞的免疫原性，从而刺激机体产生特异性的抗肿瘤免疫。

A role for shikonin-induced calreticulin on ovarian cancer cell surface in the maturation of dendritic cells

Objective It is to investigate the role of shikonin-induced apoptocic ovarian cancer cell in the immunologic mat- uration of dendritic cell （DC） and explore the related mechanisms. Methods HO -8910 cells were treated with shikonin at different concentrations for 48h. The apoptotic cells and the cell surface calreticulin on HO - 8910 cells were determined by flow cytometry. And then the pre-treated and non-pretreated HO -8910 cells were co-cultured with human peripheral blood derived DC in the presence or absence of calreticulin inhibitory peptides tor 48h respectively. DC co-cultured with HO - 8910 ceils without any medicine pretreatment was used as a negative control and the DC alone was used as a blank control. Then cell surface level of CD86 on DC were detected and analyzed by flow cytometry. Results Shikonin-treated HO -8910 cells exhibi- ted significantly higher levels of apoptosis rate and cell surface calreticulin when compared with that of non-treated HO -8910 cells （P 〈 0.01 ） in a concentration dependent manner. Further, the cell surface level of CD86 on DC was significantly elevat- ed after co-culturing with shikonin-treated HO -8910 （：ells （P 〈 0.01 ） and the CRT inhibiting peptides significantly decreased the up-regulation level of CD86. Conclusion Shikonin-induced apoptotic HO -8910 cells increased the maturation level of DC through a cell surface calreticulin-mediated interaction. It suggests that the shikonin induced immunogenic cell death of ovarian cancer cells （：an elicit protective tumor-specific immune respmlses in immunogenic chemotherapy.