Zang Guang-Hui, Li Rui, Zhou Rong-Sheng, Hao Lin, He Hou-Guang, Zhang Wen-Da, Dong Yang, Han Cong-Hui
Department of Urology, Xuzhou Central Hospital, Xuzhou, Jiangsu 221009, P.R. China.
Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430014, P.R. China.
Oncol Lett. 2018 Feb;15(2):2273-2277. doi: 10.3892/ol.2017.7589. Epub 2017 Dec 12.
The present study explored the effects of disodium cantharidinate (DC) on the peripheral blood-derived dendritic cells of patients with bladder carcinoma. The peripheral blood mononuclear cells from the 15 cases of urinary bladder carcinoma of middle and advanced stage were separated, and dendritic cells were prepared. The morphological changes of dendritic cells were observed. Flow cytometry was used to detect the expression levels of CD1a and CD83 on dendritic cell surface. MTT assay was utilized to measure the proliferation ability of allogeneic lymphocyte stimulated by DC. Annexin V-FITC/propidium iodide (PI) double staining flow cytometry method was carried out to detect cell apoptosis after treatment with DC. The changes in caspase-3 and PARP expression levels were investigated by western blot method. The high-dose DC resulted in a significant increase in the expressions of dendritic cell phenotyptic molecules CDla and CD83 as compared to control group. In addition, the proliferation index of allogenic lymphocyte stimulated by DC was significantly higher than that of control group. Moreover, MTT assay showed significant inhibition of the growth of BIU-87 cells. After 24 h of DC treatment, double staining flow cytometry confirmed the ability of DC to induce cell apoptosis. Further, western blot method showed a significant elevation of caspase-3 and PARP protein expression after DC treatment. In conclusion, DC treatment could induce dendritic cell maturation of patient with carcinoma of urinary bladder and promote its functional changes. Furthermore, DC was able to inhibit the proliferation of cell BIU-87 and also has the ability to induce cell apoptosis.
本研究探讨了斑蝥酸钠（DC）对膀胱癌患者外周血来源树突状细胞的影响。分离了15例中晚期膀胱癌患者的外周血单个核细胞，并制备树突状细胞。观察树突状细胞的形态变化。采用流式细胞术检测树突状细胞表面CD1a和CD83的表达水平。利用MTT法检测DC刺激的同种异体淋巴细胞的增殖能力。采用Annexin V-FITC/碘化丙啶（PI）双染流式细胞术检测DC处理后的细胞凋亡情况。通过蛋白质印迹法研究caspase-3和PARP表达水平的变化。与对照组相比，高剂量DC导致树突状细胞表型分子CD1a和CD83的表达显著增加。此外，DC刺激的同种异体淋巴细胞的增殖指数显著高于对照组。而且，MTT法显示对BIU-87细胞的生长有显著抑制作用。DC处理24小时后，双染流式细胞术证实了DC诱导细胞凋亡的能力。此外，蛋白质印迹法显示DC处理后caspase-3和PARP蛋白表达显著升高。总之，DC处理可诱导膀胱癌患者树突状细胞成熟并促进其功能改变。此外，DC能够抑制BIU-87细胞的增殖，并且还具有诱导细胞凋亡的能力。
