NDV/FMW对耐维罗非尼的恶性黑色素瘤细胞免疫原性死亡相关分子表达的影响

目的 探讨新城疫病毒（Newcastle disease virus, NDV）毒株FMW（NDV/FMW）对恶性黑色素瘤维罗非尼（vemurafenib）耐药细胞株WM3248/vemurafenib免疫原性死亡相关分子表达的影响。方法 通过病毒滴度检测考察病毒复制情况；采用CCK8试剂盒检测细胞存活率；Western印迹法检测细胞凋亡蛋白、细胞和上清液中高迁移率族蛋白1（HMGB1）和热休克蛋白90（HSP90）变化；流式细胞术和激光共聚焦技术检测细胞膜上钙网蛋白（calreticulin, CRT）的表达；荧光素酶法检测细胞上清液中的三磷酸腺苷（ATP）的释放。结果 NDV/FMW在WM3248/vemurafenib细胞株内复制，降低耐药细胞存活率并增加细胞凋亡（P<0.001）；NDV/FMW诱导WM3248/vemurafenib细胞膜CRT表达增加（P<0.01），ATP分泌、HMGB1和HSP90释放（P<0.05）。结论 NDV/FMW促进WM3248/vemurafenib细胞株死亡，免疫原性死亡是其死亡机制之一。

Effects of NDV/FMW on the expression of immunogenic cell death-related molecules in vemurafenib-resistant malignant melanoma cells

Objective To investigate the effects of Newcastle disease virus (NDV) strain FMW (NDV/FMW) on the expression of immunogenic cell death-related molecules in vemurafenib-resistant malignant melanoma WM3248/vemurafenib cells. Methods Human melanoma WM3248 and WM3248/vemuragenib cells were treated with NDV/FMW. Multi-step viral growth curves was used to detect virus yield; cell viability was detected by CCK8; the changes of apoptosis-related proteins and the secretion of high mobility group protein 1 (HMGB1) and heat shock protein 90 (HSP90) in the supernatant were detected by Western blotting; flow cytometry and laser copolymerization were used to detect the expression of calreticulin (CRT) on cell membrane; the release of adenosine triphosphate (ATP) in the supernatant was tested by fluorescein enzymatic method. Results NDV/FMW was duplicated in WM3248/vemurafenib cells, inhibited cell viability and promoted apoptosis (P<0.001). NDV/FMW enhanced the accumulation of CRT (P<0.01) on the cell surface and promoted the secretion of ATP, HMGB1 and HSP90 in WM3248/vemurafenib cells (P<0.05). Conclusion NDV/FMW can induce the death of WM3248/vemurafenib cells, and the immunogenic cell death is one of the mechanisms of cell death.