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Global proteomic and phosphoproteomic analysis of WT HEC-50B and MAPK14-/- HEC-50B cells cultured as 2D monolayers and 3D spheroids.

ABSTRACT: The molecular underpinnings of High Grade Endometrial Carcinoma (HGEC) metastatic growth and survival are poorly understood. Here we show that ascites-derived and primary tumor HGEC cell lines in 3D spheroid culture faithfully recapitulate key features of malignant peritoneal effusion and exhibit fundamentally distinct transcriptomic, proteomic and metabolomic landscapes when compared with conventional 2D monolayers. Using genetic screening platform we identify MAPK14 (which encodes the protein kinase p38a) as a specific requirement for HGEC in spheroid culture. MAPK14/p38a has broad roles in programing the phosphoproteome of HGEC spheroids, yet has negligible impact on monolayer cultures. Using global proteomic and phosphoproteomic analysis we identified several kinases specifically activated in 3D spheroids. The largest increases and decreases in phosphoprotein abundance caused by spheroid culture were all MAPK14-dependent. Therefore, MAPK14 plays a key role in specifying the global kinome of HGEC cells in spheroid culture.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Scott Lyons

LAB HEAD: Cyrus Vaziri

PROVIDER: PXD053011 | Pride | 2025-10-07

REPOSITORIES: Pride

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			Action	DRS
	20230505_OE480_PC1100-Vaziri_TMTpro-phospho_F.msf	Msf
	20230505_OE480_PC1100-Vaziri_TMTpro-phospho_F.pep.xml	Pepxml
	20230505_OE480_PC1100-Vaziri_TMTpro-phospho_F1-R1.raw	Raw
	20230505_OE480_PC1100-Vaziri_TMTpro-phospho_F1-R2.raw	Raw
	20230505_OE480_PC1100-Vaziri_TMTpro-phospho_F2-R1.raw	Raw

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