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Resistance to cisplatin and paclitaxel does not affect the sensitivity of human ovarian cancer cells to antiprogestin-induced cytotoxicity.

Gamarra-Luques CD, Hapon MB, Goyeneche AA, Telleria CM - J Ovarian Res (2014)

Bottom Line: The cells that did not die and repopulate the culture after the chemotherapies were termed Platinum-Taxane-EScape cells (PTES).Parental cells were compared against their PTES derivatives in their responses to further platinum-taxane treatments.Cells subjected to six pulse-selection cycles of cisplatin-paclitaxel gave rise to sibling derivatives that displayed ~2-7 fold reduction in their sensitivities to further chemotherapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South, Dakota, 414 East Clark Street, Vermillion, SD 57069, USA ; Present Address: Institute of Medicine and Experimental Biology of Cuyo, National Council for Scientific and Technical Research (CONICET), Mendoza, Argentina.

ABSTRACT

Background: Antiprogestin compounds have been shown to be effective in blocking the growth of ovarian cancer cells of different genetic backgrounds. Herein we studied the anti-ovarian cancer effect of a series of antiprogestins sharing the chemical backbone of the most characterized antiprogestin, mifepristone, but with unique modifications in position C-17 of the steroid ring. We assessed the effect of mifepristone-like antiprogestins on the growth of ovarian cancer cells sensitive to the standard combination therapy cisplatin-paclitaxel or made double-resistant upon six cycles of pulse-selection with the drugs used at clinically relevant concentrations and exposure times.

Methods: IGROV-1 and SKOV-3 cells were pulsed with 20 μM cisplatin for 1 h followed by 100 nM paclitaxel for 3 h once a week for six weeks. The cells that did not die and repopulate the culture after the chemotherapies were termed Platinum-Taxane-EScape cells (PTES). Parental cells were compared against their PTES derivatives in their responses to further platinum-taxane treatments. Moreover, both ovarian cancer cells and their PTES siblings were exposed to escalating doses of the various antiprogestin derivatives. We assessed cell growth, viability and sub-G1 DNA content using microcapillary cytometry. Cyclin-dependent kinase inhibitors p21(cip1) and p27(kip1) and cleavage of downstream caspase-3 substrate PARP were used to assess whether cell fate, as a consequence of treatment, was limited to cytostasis or progressed to lethality.

Results: Cells subjected to six pulse-selection cycles of cisplatin-paclitaxel gave rise to sibling derivatives that displayed ~2-7 fold reduction in their sensitivities to further chemotherapy. However, regardless of the sensitivity the cells developed to the combination cisplatin-paclitaxel, they displayed similar sensitivity to the antiprogestins, which blocked their growth in a dose-related manner, with lower concentrations causing cytostasis, and higher concentrations causing lethality.

Conclusions: Antiprogestins carrying a backbone similar to mifepristone are cytotoxic to ovarian cancer cells in a manner that does not depend on the sensitivity the cells have to the standard ovarian cancer chemotherapeutics, cisplatin and paclitaxel. Thus, antiprogestin therapy could be used to treat ovarian cancer cells showing resistance to both platinum and taxanes.

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Related in: MedlinePlus

Generation of ovarian cancer cells resistant to CDDP and PTX. (A) Graphical representation of the procedure performed to generate cells with lower sensitivity to both CDDP and PTX. Lighter cells represent growing cells whereas darker cells are cells that survive therapy. Cells showing nuclear fragmentation represent those dying in response to chemotherapy. Phase contrast images at lower or higher magnifications of the morphologies displayed by IGROV-1 and the IGROV-1 PTES (B) and that of SKOV-3 and SKOV-3 PTES siblings (C). Scale bar, 100 μm.
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Figure 2: Generation of ovarian cancer cells resistant to CDDP and PTX. (A) Graphical representation of the procedure performed to generate cells with lower sensitivity to both CDDP and PTX. Lighter cells represent growing cells whereas darker cells are cells that survive therapy. Cells showing nuclear fragmentation represent those dying in response to chemotherapy. Phase contrast images at lower or higher magnifications of the morphologies displayed by IGROV-1 and the IGROV-1 PTES (B) and that of SKOV-3 and SKOV-3 PTES siblings (C). Scale bar, 100 μm.

Mentions: Ovarian carcinoma IGROV-1 and SKOV-3 cells were plated into T75 cm2 culture flasks. When the culture reached 90% confluence, the cells received one chemotherapeutic challenge consisting of 20 μM CDDP for 1 h followed by 100 nM PTX for 3 h, which was repeated weekly for six weeks. Upon the repopulation following the last chemotherapeutic challenge, the cells were considered as Platinum-Taxane-EScape cells (PTES), and were trypsinized and stored in liquid nitrogen for subsequent uses. Figure 2A displays a schematic summary of the experimental procedure implemented.


Resistance to cisplatin and paclitaxel does not affect the sensitivity of human ovarian cancer cells to antiprogestin-induced cytotoxicity.

Gamarra-Luques CD, Hapon MB, Goyeneche AA, Telleria CM - J Ovarian Res (2014)

Generation of ovarian cancer cells resistant to CDDP and PTX. (A) Graphical representation of the procedure performed to generate cells with lower sensitivity to both CDDP and PTX. Lighter cells represent growing cells whereas darker cells are cells that survive therapy. Cells showing nuclear fragmentation represent those dying in response to chemotherapy. Phase contrast images at lower or higher magnifications of the morphologies displayed by IGROV-1 and the IGROV-1 PTES (B) and that of SKOV-3 and SKOV-3 PTES siblings (C). Scale bar, 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4007005&req=5

Figure 2: Generation of ovarian cancer cells resistant to CDDP and PTX. (A) Graphical representation of the procedure performed to generate cells with lower sensitivity to both CDDP and PTX. Lighter cells represent growing cells whereas darker cells are cells that survive therapy. Cells showing nuclear fragmentation represent those dying in response to chemotherapy. Phase contrast images at lower or higher magnifications of the morphologies displayed by IGROV-1 and the IGROV-1 PTES (B) and that of SKOV-3 and SKOV-3 PTES siblings (C). Scale bar, 100 μm.
Mentions: Ovarian carcinoma IGROV-1 and SKOV-3 cells were plated into T75 cm2 culture flasks. When the culture reached 90% confluence, the cells received one chemotherapeutic challenge consisting of 20 μM CDDP for 1 h followed by 100 nM PTX for 3 h, which was repeated weekly for six weeks. Upon the repopulation following the last chemotherapeutic challenge, the cells were considered as Platinum-Taxane-EScape cells (PTES), and were trypsinized and stored in liquid nitrogen for subsequent uses. Figure 2A displays a schematic summary of the experimental procedure implemented.

Bottom Line: The cells that did not die and repopulate the culture after the chemotherapies were termed Platinum-Taxane-EScape cells (PTES).Parental cells were compared against their PTES derivatives in their responses to further platinum-taxane treatments.Cells subjected to six pulse-selection cycles of cisplatin-paclitaxel gave rise to sibling derivatives that displayed ~2-7 fold reduction in their sensitivities to further chemotherapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South, Dakota, 414 East Clark Street, Vermillion, SD 57069, USA ; Present Address: Institute of Medicine and Experimental Biology of Cuyo, National Council for Scientific and Technical Research (CONICET), Mendoza, Argentina.

ABSTRACT

Background: Antiprogestin compounds have been shown to be effective in blocking the growth of ovarian cancer cells of different genetic backgrounds. Herein we studied the anti-ovarian cancer effect of a series of antiprogestins sharing the chemical backbone of the most characterized antiprogestin, mifepristone, but with unique modifications in position C-17 of the steroid ring. We assessed the effect of mifepristone-like antiprogestins on the growth of ovarian cancer cells sensitive to the standard combination therapy cisplatin-paclitaxel or made double-resistant upon six cycles of pulse-selection with the drugs used at clinically relevant concentrations and exposure times.

Methods: IGROV-1 and SKOV-3 cells were pulsed with 20 μM cisplatin for 1 h followed by 100 nM paclitaxel for 3 h once a week for six weeks. The cells that did not die and repopulate the culture after the chemotherapies were termed Platinum-Taxane-EScape cells (PTES). Parental cells were compared against their PTES derivatives in their responses to further platinum-taxane treatments. Moreover, both ovarian cancer cells and their PTES siblings were exposed to escalating doses of the various antiprogestin derivatives. We assessed cell growth, viability and sub-G1 DNA content using microcapillary cytometry. Cyclin-dependent kinase inhibitors p21(cip1) and p27(kip1) and cleavage of downstream caspase-3 substrate PARP were used to assess whether cell fate, as a consequence of treatment, was limited to cytostasis or progressed to lethality.

Results: Cells subjected to six pulse-selection cycles of cisplatin-paclitaxel gave rise to sibling derivatives that displayed ~2-7 fold reduction in their sensitivities to further chemotherapy. However, regardless of the sensitivity the cells developed to the combination cisplatin-paclitaxel, they displayed similar sensitivity to the antiprogestins, which blocked their growth in a dose-related manner, with lower concentrations causing cytostasis, and higher concentrations causing lethality.

Conclusions: Antiprogestins carrying a backbone similar to mifepristone are cytotoxic to ovarian cancer cells in a manner that does not depend on the sensitivity the cells have to the standard ovarian cancer chemotherapeutics, cisplatin and paclitaxel. Thus, antiprogestin therapy could be used to treat ovarian cancer cells showing resistance to both platinum and taxanes.

Show MeSH
Related in: MedlinePlus