Butein增强mH2A靶向GRP78蛋白通过MAPK信号通路调控黑色素瘤的生物学行为

Butein promotes the role of mH2A in MAPK signaling pathway through targeting GRP78 in regulating biological behaviors of melanoma

目的:探讨组蛋白变异体macroH2A(mH2A)和Butein共同作用GRP78蛋白通过丝裂原激活蛋白激酶(mitogen-activation protein kinase,MAPK)信号通路调控黑色素瘤的生物学行为。方法:采用免疫组织化学检测黑色素瘤和癌旁正常组织中mH2A和葡萄糖调节蛋白78(glucose regulated protein of 78,GRP78)的表达情况;分析临床资料与mH2A和GRP78表达的关系;免疫共沉淀实验检测GRP78和mH2A蛋白的相互作用;Western印迹检测mH2A和GRP78的相关作用及Butein与mH2A之间的作用。Transwell试验检测mH2A的表达对黑色素瘤细胞侵袭能力的影响;划痕试验检测mH2A的表达对黑色素瘤细胞迁移能力的影响;Western印迹检测沉默mH2A后细胞外信号调节激酶1(ERK1)和MAPK/ERK激酶(MEK)蛋白的表达。结果:与癌旁正常组织相比,黑色素瘤组织中mH2A和GRP78表达明显降低(P<0.05);Butein可以增强mH2A的表达水平(P<0.05),mH2A可以调节GRP78蛋白的表达(P<0.05);沉默mH2A促进黑色素瘤A375细胞的侵袭迁移能力,上调MEK和ERK1蛋白的表达。结论:Butein可以增强mH2A靶向GRP78蛋白通过MAPK信号通路抑制黑色素瘤细胞的迁移和侵袭行为。

Objective： To investigate the effect of mH2A and Butein on mitogen-activation protein kinase （MAPK） signaling pathway through targeting glucose regulated protein of 78 （GRP78） in regulating the biological behavior of melanoma. Methods： Immunohistochemistry was used to detect the expressions of mH2A and GRP78 in melanoma and adjacent normal tissues. The relationship between mH2A and GRP78~ and the clinicopathological data was analyzed. The relationship between GRP78 and mH2A protein was detected by immunoprecipitation assay： The protein expressions ofmH2A and GRP78 was detected by Western blot. R-he invasion ability of melanoma after silencing mH2A was detected by Transwell invasion assay. The migration ability of melanoma after sliencing mH2A was detected by wound healing assays. The protein expressions of MEK and ERK1 after silencing mH2A was detected by Western blot. Results： The expressions of mH2A and GRP78 in melanoma tissues was significantly lower than that in adjacent normal tissues （P〈0.05）. Butein enhanced the expression of mH2A （P〈0.05）. The mH2A regulated the GRP78 protein （P〈0.05）. Silencing mH2A promoted the invasion and migration of melanoma A375 cells. The MEK and ERK1 protein expressions were up-regulated