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

Ha Yeon Shin, Wookyeom Yang, Eun Ju Lee, Gwan Hee Han, Hanbyoul Cho, Doo Byung Chay, Jae-Hoon Kim

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Background Human 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. Methods HOSE 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. Results RNA-sequencing results revealed a stronger linear correlation in gene expression between IHOSE and HOSE cells (R 2 = 0.9288) than between IHOSE or HOSE cells and ovarian cancer cells (R 2 = 0.8562 and R 2 = 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. Conclusion These 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.

Original languageEnglish
Article numbere0205297
JournalPloS one
Volume13
Issue number10
DOIs
Publication statusPublished - 2018 Oct 1

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Polyomavirus Transforming Antigens
ovarian neoplasms
epithelial cells
Epithelial Cells
cell lines
antigens
Cells
Cell Line
Ovarian Neoplasms
Gene expression
cells
gene expression
RNA Sequence Analysis
sequence analysis
RNA
cadherins
Viral Tumor Antigens
Cadherins
Research
Gene Expression

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Shin, Ha Yeon ; Yang, Wookyeom ; Lee, Eun Ju ; Han, Gwan Hee ; Cho, Hanbyoul ; Chay, Doo Byung ; Kim, Jae-Hoon. / Establishment of five immortalized human ovarian surface epithelial cell lines via SV40 T antigen or HPV E6/E7 expression. In: PloS one. 2018 ; Vol. 13, No. 10.
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title = "Establishment of five immortalized human ovarian surface epithelial cell lines via SV40 T antigen or HPV E6/E7 expression",
abstract = "Background Human 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. Methods HOSE 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. Results RNA-sequencing results revealed a stronger linear correlation in gene expression between IHOSE and HOSE cells (R 2 = 0.9288) than between IHOSE or HOSE cells and ovarian cancer cells (R 2 = 0.8562 and R 2 = 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. Conclusion These 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.",
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Establishment of five immortalized human ovarian surface epithelial cell lines via SV40 T antigen or HPV E6/E7 expression. / Shin, Ha Yeon; Yang, Wookyeom; Lee, Eun Ju; Han, Gwan Hee; Cho, Hanbyoul; Chay, Doo Byung; Kim, Jae-Hoon.

In: PloS one, Vol. 13, No. 10, e0205297, 01.10.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Establishment of five immortalized human ovarian surface epithelial cell lines via SV40 T antigen or HPV E6/E7 expression

AU - Shin, Ha Yeon

AU - Yang, Wookyeom

AU - Lee, Eun Ju

AU - Han, Gwan Hee

AU - Cho, Hanbyoul

AU - Chay, Doo Byung

AU - Kim, Jae-Hoon

PY - 2018/10/1

Y1 - 2018/10/1

N2 - Background Human 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. Methods HOSE 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. Results RNA-sequencing results revealed a stronger linear correlation in gene expression between IHOSE and HOSE cells (R 2 = 0.9288) than between IHOSE or HOSE cells and ovarian cancer cells (R 2 = 0.8562 and R 2 = 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. Conclusion These 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.

AB - Background Human 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. Methods HOSE 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. Results RNA-sequencing results revealed a stronger linear correlation in gene expression between IHOSE and HOSE cells (R 2 = 0.9288) than between IHOSE or HOSE cells and ovarian cancer cells (R 2 = 0.8562 and R 2 = 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. Conclusion These 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.

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