In vitro growth regulation of endometrial carcinoma cells by tamoxifen and medroxyprogesterone acetate

doi:10.1016/0090-8258(88)90030-3

Gynecologic Oncology

Volume 30, Issue 2, June 1988, Pages 239-250

https://doi.org/10.1016/0090-8258(88)90030-3 Get rights and content

Abstract

The growth inhibitory effects of medroxyprogesterone acetate (MPA) and tamoxifen (TAM) were tested on three long-established endometrial carcinoma cell lines (HEC-1, KLE, and RL95-2) and on UM-EC-1, a new endometrial carcinoma cell line established in our laboratory. MPA and TAM were used in growth experiments either alone, simultaneously, or sequentially. The MCF-7 breast cancer cell line was used as a control. None of the endometrial carcinoma cell lines showed significant sensitivity to 0.1–10 μM MPA. In contrast, 10 days exposure to 5 μM TAM induced 83 and 70% growth inhibition in HEC-1 and KLE cultures, whereas the growth of UM-EC-1 was inhibited by 99.7% and RL95-2 cultures by 100%. TAM-induced growth inhibition was reversible since all cell lines resumed logarithmic growth when TAM was removed from the culture medium. Addition of 17-β-estradiol (E₂) to the culture medium did not accelerate recovery, and reversal of TAM-induced growth inhibition was not seen when TAM and E₂ were added simultaneously. This is consistent with our finding that, except for MCF-7, these cell lines did not show detectable estrogen receptor (ER) activity in assays performed at the time of these experiments. When treated sequentially with TAM and MPA, all cell lines resumed logarithmic growth when medium containing TAM was replaced with medium containing MPA. Simultaneous exposure to 5 μM MPA and 5 μM TAM resulted in a slight additive growth inhibitory effects only in KLE cultures. Our results show that MPA does not have growth inhibitory effects in these endometrial carcinoma cell cultures, whereas TAM exerts a potent growth inhibitory effect that is not reversed by estrogen and may thus be mediated through a mechanism different from blockade of ER. In vitro results with the UM-EC-1 cell line correlated with the clinical response of the cell line donor. Her disease progressed during postoperative MPA therapy, but subsequently she responded to TAM therapy.

References (46)

E.I. Kohorn
Gestagens and endometrial carcinoma
Gynecol. Oncol.
(1976)
M.A. Quinn et al.
Endometrial carcinoma: Steroid receptors and response to medroxyprogesterone acetate
Gynecol. Oncol.
(1985)
C.E. Ehrlich et al.
Cytoplasmic progesterone and estradiol receptors in normal, hyperplastic and carcinomatous endometria: Therapeutic implications
Amer. J. Obstet. Gynecol.
(1981)
J. Bonte et al.
Tamoxifen as a possible chemotherapeutic agent in endometrial adenocarcinoma
Gynecol. Oncol.
(1981)
V. Isomaa et al.
Regulation of cytosol and nuclear progesterone receptors in rabbit uterus by estrogen, antiestrogen and progesterone administration
Biochim. Biophys. Acta
(1979)
J.A. Carlson et al.
Tamoxifen and endometrial carcinoma: Alterations in estrogen and progesterone receptors in untreated patients and combination hormonal therapy in advanced neoplasia
Amer. J. Obstet. Gynecol.
(1984)
P.E. Schwartz et al.
Tamoxifen-induced increase in cytosol progestin receptor levels in a case of metastatic endometrial cancer
Gynecol. Oncol.
(1983)
H. Kuramoto et al.
Establishment of a cell line of human endometrial adenocarcinoma in vitro
Amer. J. Obstet. Gynecol.
(1972)
G.S. Richardson et al.
KLE: A cell line with defective estrogen receptor derived from undifferentiated endometrial cancer
Gynecol. Oncol.
(1984)
S. Grenman et al.
In vitro response of cervical cancer cell lines CaSki, HeLa, and ME-180 to the antiestrogen tamoxifen
Gynecol. Oncol.
(1988)

P.G. Satyaswaroop et al.

Metabolism and effects of progesterone in the human endometrial adenocarcinoma cell line HEC-1

Steroids

(1980)

C.F. Holinka et al.

Responses to estradiol in a human endometrial adenocarcinoma cell line (Ishikawa)

J. Steroid Biochem.

(1986)

M.H. Briggs et al.

The treatment of cancer by progestogens

Hosp. Med.

(1967)

M. Gr̈onroos et al.

Steroid receptors and response of endometrial cancer to hormones

Ann. Chir. Gynecol.

(1987)

W.T. Creasman et al.

Clinical correlates of estrogen- and progesterone-binding proteins in human endometrial adenocarcinoma

Obstet. Gynecol.

(1982)

A.J.I. Kauppila et al.

Prediction of clinical outcome with estrogen and progestin receptor concentrations and their relationship to clinical and histopathological variables in endometrial cancer

Cancer Res.

(1986)

M. Neumannova et al.

Short-term effects of tamoxifen, medroxprogesterone acetate and their combination on receptor kinetics and 17-β-hydroxysteroid dehydrogenase in human endometrium

Obstet. Gynecol.

(1985)

O. Janne et al.

Female sex steroid receptors in normal, hyperplastic and carcinomatous endometrium: The relationship to serum steroid hormones and gonadotropins and changes during medroxprogesterone acetate administration

Int. J. Cancer

(1979)

P. Biondani et al.

Endometrial cancer: Steroid receptors and medroxyprogesterone acetate treatment

K.D. Swenerton et al.

Treatment of advanced endometrial carcinoma with tamoxifen

N. Engl. J. Med.

(1979)

A. Kauppila et al.

Case report: Endometrial carcinoma insensitive to progestin and cytotoxic chemotherapy may respond to tamoxifen

Acta Obstet. Gynecol. Scand.

(1981)

M. Slavik et al.

Phase II clinical study of tamoxifen in advanced endometrial adenocarcinoma: A gynecologic oncology group study

Cancer Treat. Rep.

(1984)

K.D. Swenerton et al.

Efficacy of tamoxifen in endometrial cancer

Cited by (29)

Aberrant upregulation of CDK1 contributes to medroxyprogesterone acetate (MPA) resistance in cancer-associated fibroblasts of the endometrium
2022, Biochemical and Biophysical Research Communications
Citation Excerpt :
Hence, abnormalities along PR signaling axis may lead to the development of progesterone resistance [15]. Most studies focused on the epithelial tumor cells when investigating the effect of progesterone in cancer therapy [12,16–18], while largely ignoring the role of fibroblasts. In the normal endometrium, fibroblasts promote epithelial cell development and differentiation [19,20].
The response to medroxyprogesterone acetate (MPA) decreases as endometrial disease progresses from the benign to malignancy. In a mouse model, progesterone receptor (PR) expression in normal fibroblasts is accountable for the MPA's inhibitory effects in cancer cells. However, it is still unclear, if and how, fibroblasts from human tumors respond to MPA. In this study, three benign-associated fibroblasts (BAFs) and four cancer-associated fibroblasts (CAFs) were isolated from human benign and cancerous endometrial tissues, respectively, to examine MPA activation on PR signaling. PR-B protein expression were heterogeneously expressed in both CAFs and BAFs, despite a lower mRNA expression in the former. In a luciferase reporter assay, MPA treatment stimulated some PR DNA-binding activity in BAFs but not in CAFs. Yet, activation of PR target gene was generally more pronounced in MPA-treated CAFs compared to BAFs. Cyclin-dependent kinase 1 (CDK1) was exclusively upregulated by 10 nM MPA in CAFs (5.1-fold vs. 1.1-fold in BAFs, P < 0.05), leading to a higher CDK1 protein expression. Subsequently in a dose-response study, CAFs showed an average of ∼20% higher cell viability when compared to BAFs, indicative of drug resistance to MPA. MPA resistance was also observed in EC-CAFs co-culture, when MPA-treated cells showed greater tumor spheroid formation than in EC-BAFs co-culture (2-fold, P < 0.01). The increased cell viability observed in CAFs was reversed with mifepristone (RU486), a PR antagonist which suppressed MPA-induced CDK1 expression. This indicates that MPA-induced abnormal upregulation of CDK1 may contribute to the enhanced CAFs cell proliferation, suggesting a new mechanism of MPA resistance within endometrial cancer microenvironment.
Establishment and characterization of a poorly differentiated lethal human endometrial carcinoma cell line (NOU-1) with karyotype 46,XX
2002, Cancer Genetics and Cytogenetics
Immortal gynecologic cell lines, especially those of endometrial origin are diverse with multiple chromosomal alterations, making the study of origin and molecular genetic characteristics of such neoplasms difficult, if not impossible. We have established a new epithelial, poorly differentiated endometrial adenocarcinoma cell line (NOU-1) with a 46,XX chromosome complement. To confirm the tumor characteristics, we injected this cell line subcutaneously into the nude mouse. The tumor grown in vivo had the identical histology and the same 46,XX chromosome complement as the original tumor excised from the patient. The cells from the original tumor and those in the nude mouse shared the same characteristics with respect to histopathology, immunohistochemistry, steroid hormone receptor content, and growth characteristics. We report on the only established cell line of a highly aggressive, poorly differentiated endometrial adenocarcinoma with 46,XX chromosome complement where both estrogen and progesterone receptors are absent.
Triage of abnormal postmenopausal bleeding: A comparison of endometrial biopsy and transvaginal sonohysterography versus fractional curettage with hysteroscopy
1998, American Journal of Obstetrics and Gynecology
OBJECTIVE: We sought to compare the combined diagnostic reliability of sonohysterography and endometrial biopsy with fractional curettage with hysteroscopy in the initial evaluation of postmenopausal women with abnormal uterine bleeding. STUDY DESIGN: This year-long, prospective, controlled, clinical investigation was initiated Sept. 1, 1995. All postmenopausal women with abnormal uterine bleeding were offered inclusion, with 104 enrolled. An endometrial biopsy was performed at the time of initial evaluation. Routine transvaginal ultrasonography was then used to measure the uterus, ovaries, and endometrial stripe thickness, followed immediately by sonohysterography to evaluate the symmetry of endometrial wall thickness and delineate any intraluminal masses. Definitive histopathologic sampling was obtained by fractional curettage with hysteroscopy and statistically compared with the diagnoses arrived at by endometrial biopsy and sonohysterography. RESULTS: The combination of endometrial biopsy and transvaginal sonohysterography positively correlated with the surgical findings >95% of the time, with a sensitivity and specificity of 94% and 96%, respectively (confidence interval 91% to 99%). No patients with endometrial hyperplasia or cancer were misdiagnosed. CONCLUSIONS: Sonohysterography combined with endometrial biopsy is a reliable office tool for evaluating postmenopausal women with abnormal uterine bleeding. Medical management of those patients identified as having no endometrial abnormalities can be considered with confidence, while saving the cost and surgical risk of fractional curettage. Patients with intraluminal masses should be referred for surgical management in a timely fashion. (Am J Obstet Gynecol 1998;178:956-61.)
Gynaecological aspects of tamoxifen treatment in breast cancer patients
1997, Cancer Treatment Reviews
A retrospective study of tamoxifen and endometrial cancer in breast cancer patients
1995, Gynecologic Oncology
Objective. To assess the relationship between low-dose tamoxifen, usage and endometrial cancer in breast cancer patients. Methods. In this case-control study, the records of the 1017 patients treated at Wilford Hall Medical Center for primary breast cancer between 1978 and 1989 were reviewed. Dose and duration of tamoxifen therapy were recorded as well as the presence of a uterus. Potential confounding variables including diabetes mellitus, hypertension, age, weight, tobacco use, and family history of breast or gynecologic cancer were recorded. Results. Of the 1017 patients in the study, 56 had inadequate records and 375 had undergone a prior hysterectomy and these patients were excluded from the study, leaving 586 eligible patients. Of these 586 women with primary breast cancer and no previous hysterectomy, 108 patients had received tamoxifen, 10 mg twice a day for greater than 1 year. Four of the 108 patients subsequently developed endometrial adenocarcinoma. Four hundred seventy-eight breast cancer patients did not receive tamoxifen and 4 later developed endometrial adenocarcinoma. The odds ratio for the development of endometrial cancer when exposed to tamoxifen was 15.2 with a 95% confidence interval of 2.8–84.4. The patients exposed and not exposed to tamoxifen were compared for potential confounding variables and after controlling for the potential confounders, tamoxifen was found to be an independent risk factor for the development of endometrial cancer. Conclusions. Low-dose tamoxifen when given for greater than 1 year is associated with secondary endometrial cancer in patients with primary breast cancer.
Characterization and radiosensitivity of UT-EC-2A and UT-EC-2B, two new highly radiosensitive endometrial cancer cell lines derived from a primary and metastatic tumor of the same patient
1995, Gynecologic Oncology
UT-EC-2A was established from a patient with moderately differentiated Stage III endometrial adenocarcinoma with squamous metaplasia. UT-EC-2B was established from the same patient 17 months later from a metastasis in the left supraclavicular fossa. The origin of these cell lines was confirmed by DNA identity testing. Nude mice tumors produced by UT-EC-2A and UT-EC-2B cells recapitulated the histology of the original human tumors. Flow cytometric DNA contents of both primary and metastatic human tumors as well as corresponding nude mice tumors were diploid. The S-phase fractions of both cell lines were ⩾30%. The UT-EC-2A cell line was cytogenetically normal. The UT-EC-2B cell line had quite simple karyotype at low passage with an extra i(18p) and a deletion 21q, but at higher passages an additional three-way translocation 5;14;19 was observed. Radiosensitivity of the cell lines was tested with the 96-well plate clonogenic assay. The areas under the survival curves corresponding to the mean inactivation doses of UT-EC-2A and UT-EC-2B were 0.65 ± 0.10 and 0.60 ± 0.06 Gy, respectively. Measured survival at 2.0 Gy (SF2) was 0.042 for UT-EC-2A, 0.044 for UT-EC-2B, and 0.2 for skin fibroblasts. These cell lines are among the most radiosensitive human cancer cell lines described in the literature. Studying the characteristics of primary and metastatic cells derived from the same patient provides an opportunity to evaluate tumor progression.

View all citing articles on Scopus

^☆: Supported by USPHS Grant CA 28564 from the National Cancer Institute and a grant from the Finnish Cancer Society.

^☆☆: Presented at the 15th Annual Meeting of the Western Association of Gynecologic Oncologists (WAGO) May 13–16, 1987, in Tucson, AZ.

³: Present address: Department of Obstetrics and Gynecology at the University of Turku, Turku, Finland.

View full text

Regular articleIn vitro growth regulation of endometrial carcinoma cells by tamoxifen and medroxyprogesterone acetate☆,☆☆

Abstract

Gynecol. Oncol.

Gynecol. Oncol.

Amer. J. Obstet. Gynecol.

Gynecol. Oncol.

Biochim. Biophys. Acta

Amer. J. Obstet. Gynecol.

Gynecol. Oncol.

Amer. J. Obstet. Gynecol.

Gynecol. Oncol.

Gynecol. Oncol.

Steroids

J. Steroid Biochem.

The treatment of cancer by progestogens

Hosp. Med.

Steroid receptors and response of endometrial cancer to hormones

Ann. Chir. Gynecol.

Clinical correlates of estrogen- and progesterone-binding proteins in human endometrial adenocarcinoma

Obstet. Gynecol.

Prediction of clinical outcome with estrogen and progestin receptor concentrations and their relationship to clinical and histopathological variables in endometrial cancer

Cancer Res.

Short-term effects of tamoxifen, medroxprogesterone acetate and their combination on receptor kinetics and 17-β-hydroxysteroid dehydrogenase in human endometrium

Obstet. Gynecol.

Female sex steroid receptors in normal, hyperplastic and carcinomatous endometrium: The relationship to serum steroid hormones and gonadotropins and changes during medroxprogesterone acetate administration

Int. J. Cancer

Endometrial cancer: Steroid receptors and medroxyprogesterone acetate treatment

Treatment of advanced endometrial carcinoma with tamoxifen

N. Engl. J. Med.

Case report: Endometrial carcinoma insensitive to progestin and cytotoxic chemotherapy may respond to tamoxifen

Acta Obstet. Gynecol. Scand.

Phase II clinical study of tamoxifen in advanced endometrial adenocarcinoma: A gynecologic oncology group study

Cancer Treat. Rep.

Efficacy of tamoxifen in endometrial cancer

Regular article
In vitro growth regulation of endometrial carcinoma cells by tamoxifen and medroxyprogesterone acetate☆,☆☆