Fig 1.
H&E and ER IHC staining of VOA1066 samples.
Both the primary tumor and xenografted subcutaneous and intraperitoneal tumors of VOA1066 showed minimal focal or no staining of Mullerian epithelial markers ER, CK7, EMA and PAX8.
Table 1.
STR analysis results of the parental tumor and VOA1066 cell line.
Fig 2.
The proliferation and morphology of VOA1066 cells in vitro and in vivo.
(A) A representative image of VOA1066 cells on petri dish. (B) Growth curves of VOA1066 in different culture media. (C) Subcutaneous tumor growth curve of VOA1066 cells. (D) Representative images of intraperitoneal xenografted tumors and their metastasis of VOA1066 cells.
Fig 3.
Analysis of 20 metaphases of VOA1066 cells revealed that VOA1066 cells have a stable diploid genome (A) with a translocation between chromosomes 3 and 14: 46,XX, t(3;14)(p25;q21) (B).
Fig 4.
Mutations and protein expression of ARID1A and ARID1B genes in VOA1066.
(A) Whole exome sequencing revealed shared somatic mutations between primary tumor and cell line, including two inactivating mutations of ARID1A, two inactivating mutations of ARID1B and a RNase IIIb domain hotspot mutation of DICER1. (B) IHC staining of ARID1A and ARID1B VOA1066 samples. (C) Western blotting analysis of SWI/SNF proteins in VOA1066 and HEC50 cells. (D) Representative image of SMARCA4 staining in VOA1066 primary tumor. (E) Co-immunoprecipitation analysis of SWI/SNF protein interactions in VOA1066 and HEC50 cells. (F) Western blotting analysis of PTEN and pAKT-473 levels in VOA1066 and HEC50 cells.
Fig 5.
Mutations of DICER1 genes in DDEC.
(A) Sanger sequencing validated the DICER1 p.D1810Y mutation in both DNA and RNA samples of VOA1066 cells. (B) MicroRNA analyses in VOA1066 cells and SVOG3e cells, a DICER1-wildtype granulosa cell line. (C) Mutational analysis of DICER1 RNase IIIb domain in additional 33 DDEC cases. (D) IHC analysis of SWI/SNF proteins (SMARCA4, SMARCB1, ARID1A, ARID1B) in 6 cases of endometrial cancer with DICER1 RNase IIIb domain hotspot mutations at Vancouver General Hospital (VGH) and mutational analyses of 17 cases of TCGA endometrial cancers with DICER1 RNase IIIb domain hotspot mutations.