髓样分化蛋白2通过调控EGFR信号通路影响三阴性乳腺癌紫杉醇耐药性

Myeloid differentiation protein 2 affects paclitaxel resistance in triple-negative breast cancer by regulating EGFR signaling pathway

目的探讨髓样分化蛋白2(myeloid differentiation protein-2,MD-2)通过EGFR信号通路对三阴性乳腺癌(triple negative breast cancer,TNBC)细胞紫杉醇耐药性的影响。方法采用免疫组织化学法检测TNBC患者癌组织和癌旁组织中MD-2的表达并分析MD-2表达与患者临床病理参数的关系。构建TNBC紫杉醇耐药细胞株,干预细胞中MD-2的表达。Transwell检测细胞侵袭,流式细胞术检测细胞凋亡。转录组测序筛选MD-2调控的信号通路,并通过Western blot验证。结果相较于癌旁组织,癌组织中MD-2表达显著增强。MD-2的高表达与临床分期、肿瘤大小、肿瘤复发转移等情况密切相关(χ^(2)=4.50,P=0.032;χ^(2)=2.55,P=0.011;χ^(2)=4.40,P=0.036)。细胞实验中,与正常乳腺细胞比较,TNBC细胞系中MD-2表达显著增强。与sh-NC组比较(100±11.52)(6.81±0.57),敲减MD-2能抑制紫杉醇耐药TNBC细胞的侵袭(61.44±6.78)(t=4.99,P=0.008)、促进凋亡(15.19±1.06)(t=12.06,P<0.001)。转录组测序以及Western blot检测结果显示,MD-2主要通过对EGFR信号通路进行调控进而影响TNBC细胞的生物学行为。结论MD-2可促进TNBC细胞侵袭及紫杉醇耐药,该作用可能通过影响EGFR信号通路实现,MD-2有望成为TNBC治疗中的有效靶点。

Objective To investigate the effects of myeloid differentiation protein-2(MD-2)on paclitaxel resistance cells in triple negative breast cancer(TNBC)through EGFR signaling pathway.Methods Immunohistochemical method was used to detect the expression of MD-2 in cancer tissue and adjacent tissue of TNBC patients,and the relationship between MD-2 expression and clinicopathological parameters of patients was analyzed.The TNBC paclitaxel-resistant cell line was constructed and MD-2 expression in cells was interfered.Cell invasion was detected by Transwell,and cell apoptosis was detected by flow cytometry.The signaling pathways regulated by MD-2 were screened by transcriptome sequencing and verified by Western blot.Results The expression of MD-2 was significantly enhanced in cancer tissues relative to adjacent tissues.High expression of MD-2 was closely related to clinical stage,tumor size,tumor recurrence and metastasis(χ^(2)=4.50,P=0.032;χ^(2)=2.55,P=0.011;χ^(2)=4.40,P=0.036).In cell experiments,compared with normal breast cells,the expression of MD-2 in TNBC cell lines was significantly enhanced.Compared with sh-NC group(100±11.52)(6.81±0.57),knockdown of MD-2 could inhibit the invasion(61.44±6.78)(t=4.99,P=0.008)but promote apoptosis(15.19±1.06)(t=12.06,P<0.001)of paclitaxel resistant TNBC cells.Transcriptome sequencing and Western blot results showed that MD-2 mainly affects the biological behavior of TNBC cells by regulating the EGFR signaling pathway.Conclusions MD-2 promoted TNBC cell invasion and paclitaxel resistance,which may be achieved by affecting the EGFR signaling pathway.MD-2 is expected to become an effective target in TNBC treatment.