Establishment of a human ovarian endometrioid carcinoma cell line by constitutive expression of cyclin-dependent kinase 4, cyclin D1 and telomerase reverse transcriptase

Abstract

Only a few human ovarian endometrioid carcinoma cell lines are currently available, partly due to the difficulty of establishing cell lines from low-grade cancers. Here, using a cell immortalization strategy consisting of i) inactivation of the p16 INK4a -pRb pathway by constitutive expression of mutant cyclin-dependent kinase 4 (R24C) (CDK4 R24C ) and cyclin D1, and ii) acquisition of telomerase reverse transcriptase (TERT) activity, we established a human ovarian endometrioid carcinoma cell line from a 46-year-old Japanese woman. That line, designated JFE-21, has proliferated continuously for over 6 months with a doubling time of ~ 55 h. JFE-21 cells exhibit polygonal shapes and proliferate without contact inhibition to form a monolayer in a jigsaw puzzle-like arrangement. Ultrastructurally, JFE-21 cells exhibit well-developed rough endoplasmic reticulum, mitochondria and lysosomes in the cytoplasm, with cells contacting each other via desmosomes. G-band karyotype analysis indicated that cells had a near-tetraploid karyotype. Immunofluorescence staining revealed that the expression profile of a series of ovarian carcinoma markers in JFE-21 cells was consistent with ovarian endometrioid carcinoma. Moreover, Sanger sequencing of DNA polymerase ε ( POLE ) gene and immunohistochemical analysis of mismatch repair (MMR) proteins revealed that JFE-21 cells were classified as the no specific molecular profile (NSMP) subtype. In addition, JFE-21 cells were sensitive to paclitaxel and carboplatin administered to the donor as therapy. These findings indicate that constitutive expression of CDK4 R24C , cyclin D1 and TERT genes may be an option to establish cell lines from low-grade cancers, including ovarian endometrioid carcinoma.

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