Disruption of the retinoblastoma pathway by small interfering RNA and ectopic expression of the catalytic subunit of telomerase lead to immortalization of human ovarian surface epithelial cells.
The risk of developing ovarian cancer is about 1% over a lifetime, but it is the most deadly gynecologic cancer, in part due to lack of diagnostic markers for early-stage disease and cell model system for studying early neoplastic changes. Most existing immortal human ovarian surface epithelial cells were achieved by using viral protein such as SV40 T/t antigen or E6/E7, which inactivate multiple cellular pathways. In the current study, we used a small interfering RNA (siRNA) against the retinoblastoma gene (pRb) and ectopic expression of human telomerase reverse transcriptase (hTERT) to immortalize the primary ovarian epithelial cell line OSE137 and two additional human ovarian surface epithelial cells. The immortalized OSE137 showed increased telomerase activity, lengthened telomeres, increased G2/M phase, altered cell-cycle regulatory proteins but nontumorigenic. As both Rb and hTERT pathways are commonly altered in human ovarian cancer and these genetic changes are faithfully modeled in these cells without using viral protein, these immortal cells represent an authentic in vitro model system with which to study the initiation and progression of human ovarian cancer.
Yang
G
G
Department of Pathology, The University of Texas MD Anderson Cancer Center, Holcombe Boulevard, Houston, TX 77030, USA.
Rosen
D G
DG
Colacino
J A
JA
Mercado-Uribe
I
I
Liu
J
J
D001483:Base Sequence; D020134:Catalytic Domain; D002461:Cell Line, Transformed; D004847:Epithelial Cells; D005260:Female; D006801:Humans; D007621:Karyotyping; D008954:Models, Biological; D008969:Molecular Sequence Data; D010051:Ovarian Neoplasms / Q000188:drug therapy / Q000378:metabolism; D010053:Ovary; D034741:RNA, Small Interfering / Q000494:pharmacology*; D016160:Retinoblastoma Protein / Q000235:genetics / Q000378:metabolism*; D019098:Telomerase / Q000378:metabolism*
D034741:RNA, Small Interfering; D016160:Retinoblastoma Protein; C509186:TERT protein, human; D019098:Telomerase