目的：观察过表达Gasdermin蛋白E（gasdermin E, GSDME）基因的结直肠癌Lovo细胞经奥沙利铂处理后发生细胞焦 亡的现象。方法：通过qPCR检测结直肠癌细胞株Lovo、正常结直肠上皮细胞HCOEPIC中GSDME基因的表达水平，构建过表 达野生型和突变型GSDME的GSDME-WT和GSDME-D270A重组质粒，包装为相应慢病毒后感染Lovo细胞构建GSDME稳定 高表达细胞株，Western blotting检测WT、 D270A和空载体组中GSDME表达水平。用不同浓度奥沙利铂（0、 4、 8、16、32、64 μg/ ml）作用于WT、 D270A组Lovo 细胞和 HCOEPIC 细胞，显微镜下观察细胞的形态变化。结果：GSDME在HCOEPIC 细胞中表 达显著高于Lovo细胞（P<0.01）。成功构建GSDME-WT和GSDME-D270A高表达重组质粒和相应的Lovo细胞株，与空载体组 相比，WT和D270A组Lovo细胞中GSDME表达水平明显升高（均P<0.05）。镜下观察发现， 64 μg/ml奥沙利铂处理各组细胞9和 12 h时，WT组的Lovo细胞和HCOEPIC细胞体积逐渐增大并向一侧“吹泡”，表现出明显的细胞焦亡现象，细胞焦亡率显著高于 无奥沙利铂处理的对照组[Lovo 细胞：（7.405±1.010） % vs（3.441±0.401）% ， P<0.05；HCOEPIC 细胞： (7.203±1.020)% vs (4.201±0.302)%， P<0.05]。结论：奥沙利铂促进过表达GSDME基因的结直肠癌Lovo细胞发生细胞焦亡。

Objective: To observe the pyrolysis of colorectal cancer Lovo cells overexpressing Gasdermin E (GSDME) after the treatment with oxaliplatin. Methods: The expression level of GSDME gene in colorectal cancer Lovo cells and normal colorectal epithelial HCOEPIC cells was detected by qPCR. The GSDME-WT (wild-type GSDME) and GSDME-D270A (mutant GSDME) recombinant plasmids were constructed. The plasmids were packaged as lentivirus and then transfected into Lovo cells to construct Lovo cell line with stable and high expression of GSDME. Western blotting was used to detect the expression level of GSDME in cells of WT, D270A and empty vector groups. Different concentrations of oxaliplatin (0, 4, 8, 16, 32, 64 μg/ml) were applied to treat Lovo cells and HCOEPIC cells in WT and D270Agroups, and the morphological changes of the cells were observed under a microscope. Results: The expression of GSDME in HCOEPIC cells was significantly higher than that in Lovo cells (P<0.01). GSDME-WT and GSDME-D270A plasmids with high GSDME expression and the corresponding Lovo cell lines were successfully constructed. Compared with the empty vector group, the expression level of GSDME in Lovo cells of WT and D270A groups were significantly increased (all P<0.05). Observation under the microscope showed that after being treated with 64 μg/ml oxaliplatin for 9 and 12 hours, the volume of Lovo cells and HCOEPIC cells in WT group gradually increased and“blistered”to one side and showed obvious pyrolysis phenomenon. The pyrolysis rate of cells in WT group was significantly higher than that of the control group without oxaliplatin treatment (Lovo cells: [7.405± 1.010]% vs [3.441±0.401]%, P<0.05; HCOEPIC cells: [7.203±1.020]% vs [4.201±0.302]%, P<0.05). Conclusion: Oxaliplatin pro-motes the pyrolysis of colorectal cancer Lovo cells overexpressing GSDME gene.

天津市卫生计生委、天津市中医药管理局重点项目（No.2017057）；国家重点实验室开放研究课题项目（No.2018094）；天津市科技支 撑重点研发计划项目（No.19YFZCSY00420）