Establishment of five immortalized human ovarian surface epithelial cell lines via SV40 T antigen or HPV E6/E7 expression

Abstract

BackgroundHuman ovarian surface epithelial (HOSE) cells are a critical cell source for ovarian cancer research; however, they are difficult to obtain and maintain under standard laboratory conditions in large quantities. The aim of this study was to generate immortalized HOSE (IHOSE) cells with maintained properties to the original cell source, thereby guaranteeing a sufficiently large cell quantity for ovarian cancer research.MethodsHOSE cells isolated from four non-cancer patients and five IHOSE cell lines were established by induction of HPV-E6/E7 expression or SV40 large T antigen using a lenti-viral system. Each of IHOSE cells was confirmed to be distinct by STR profiling. RNA-sequencing was used to compare gene expression profiles in HOSE, IHOSE and ovarian cancer cells.ResultsRNA-sequencing results revealed a stronger linear correlation in gene expression between IHOSE and HOSE cells (R2 = 0.9288) than between IHOSE or HOSE cells and ovarian cancer cells (R2 = 0.8562 and R2 = 0.7982, respectively). The gene expression pattern of 319 differentially expressed genes revealed minimal differences between HOSE and IHOSE cells, while a strong difference between ovarian cancer cells and HOSE or IHOSE cells was observed. Furthermore, the five IHOSE cell lines displayed morphological characteristics typical of epithelial cells but showed a lower level of EpCAM, CD133 and E-cadherin, as cancer stem marker, than ovarian cancer cells. Moreover, unlike cancer cells, IHOSE cells could not form colonies in the anchorage-independent soft agar growth assay.ConclusionThese findings demonstrate that five newly established IHOSE cell lines have characteristics of progenitor HOSE cells while exhibiting continuous growth, and thus, should be highly useful as control cells for ovarian cancer research.

Metformin Reduces NGF-Induced Tumour Promoter Effects in Epithelial Ovarian Cancer Cells

“…It is interesting to note that we worked here with three different cell lines, which represent different stages in epithelial ovarian cancer progression. HOSE cells are a well-known model of immortalized, but non-tumoral ovarian surface epithelial cells [ 7 , 65 , 66 ], while the A2780 cell line was obtained from primary ovarian adenocarcinoma of a patient without treatment [ 49 ], and finally the SKOV3 cell line was isolated from the ascites of a woman with EOC [ 50 ]. SKOV3 cells are resistant to several cytotoxic drugs and are highly migratory and invasive [ 51 , 67 ], similar to metastatic cells.…”

“…That may be because HOSE are not entirely normal, but rather immortalized epithelial ovarian cells [ 66 ]. HOSE cells are significantly different from EOC cells and they are considered as non-tumoral model or used as a model of early stages of tumoral development [ 7 , 65 , 66 ]. That said, it is highly likely that they have different sets of miRs compared to primary epithelial ovarian cells.…”

Establishment of an immortalized human subglottic epithelial cell line

“…SV40 T antigen has been shown to be one of the most reliable agents for the immortalization of cells in culture, and the mechanism has been well documented . Multiple other human epithelial subsites have been immortalized utilizing SV40, including esophageal, corneal, ovarian, and urinary tract . The technique has also been applied to nonepithelial cells, such as neural, epidermal, and endothelial cells, in humans and other mammals …”

ALDH1A2 Is a Candidate Tumor Suppressor Gene in Ovarian Cancer

“…Six ovarian cancer cell lines (YDOV-139, YDOV-157, YDOV-161, YDOV-13, YDOV-105, and YDOV-151) and four HOSE cell lines (HOSE 198, 209, 211, and 213), which had been previously established [24], were maintained in a 1:1 mixture of Medium 199 and MCDB 105 (Sigma, St. Louis, MO, USA) supplemented with 10% FBS (Gemini Bio-Products, Calabasas, CA, USA). Immortalized HOSE8695 (IHOSE8695) cells had been established previously [38] and were maintained in DMEM supplemented with 10% FBS (HyClone, UT, USA) and 1% antibiotics (Cellgro, Manassas, VA, USA). All cell lines were incubated at 37 °C in a humidified incubator with 5% CO 2 .…”