<p>Expression of NR1H3 in endometrial carcinoma and its effect on the proliferation of Ishikawa cells in vitro</p>

Impaired mismatch repair (MMR) is reportedly crucial in the early stages of endometrial carcinogenesis. Although estrogen exposure is considered an important risk factor for endometrial carcinoma, the relationship between estrogen and MMR activity remains undetermined. The present study was undertaken to elucidate the effect of estrogen on MMR activity in normal and malignant endometrial cells. The expression of MMR proteins, hMLH1 and hMSH2, and its correlation with estrogen was examined using immunohistochemical and immunofluorescent techniques. The effect of estradiol (E2) on the expression of hMLH1/hMSH2 protein/mRNA and in vitro MMR activity using two types of heteroduplex (G/T mismatches, 2-base insertion-deletion loops) was examined in cultured normal endometrial glandular cells and estrogen receptor-positive endometrial carcinoma Ishikawa cells. Immunohistochemical expression of hMLH1 and hMSH2 in normal endometrial glands was positively correlated with the serum E2 levels. The expression of hMLH1/hMSH2 protein and mRNA was increased in normal endometrial glandular and Ishikawa cells by E2 treatment. In vitro MMR activity was up-regulated by E2 in both types of cell and heteroduplex. Immunofluorescent analysis demonstrated that E2 enhanced proliferation and hMLH1/hMSH2 expression in both cells; however, proliferating cells without hMLH1/hMSH2 expressions implying high-risk cells were more frequently observed under low E2 concentrations. Collectively, the E2-induced up-regulation of MMR activity in endometrial cells suggests that high estrogen levels act as an intrinsic defense against endometrial carcinogenesis, whereas the imbalance between cell growth and MMR under low E2 environment as seen at postmenopause is vulnerable to carcinogenesis.

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Hormonal modulation of ishikawa cells during three-dimensional growth in vitro☆

Journal of the Society for Gynecologic Investigation ◽

10.1016/s1071-5576(98)00015-x ◽

1998 ◽

Vol 5 (4) ◽

pp. 217-223 ◽

Cited By ~ 4

Author(s):

D PINELLI ◽

J DRAKE ◽

M WILLIAMS ◽

D CAVANAGH ◽

J BECKER

Keyword(s):

Three Dimensional ◽

Ishikawa Cells ◽

Hormonal Modulation ◽

Dimensional Growth

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Bu Shen Yang Xue Prescription Has Treating Effect on Endometrial Cancer through FSH/PI3K/AKT/Gankyrin/HIF-α/cyclinD 1 Pathway in Ishikawa Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/8412984 ◽

2018 ◽

Vol 2018 ◽

pp. 1-10

Author(s):

Yue-qun Chen ◽

Hua-li Fei ◽

Hong-li Zhu

Keyword(s):

Endometrial Cancer ◽

Endometrial Carcinoma ◽

Cancer Cell ◽

Nude Mice ◽

Follicle Stimulating Hormone ◽

Control Group ◽

Model Group ◽

Migration Ability ◽

Ishikawa Cells ◽

And Migration

Background. The formulation of Bu Shen Yang Xue (BSYX) has been clinically used in treating gynecologic disease in China, especially for the development of the endometrium. Endometrial carcinoma is the most common malignant tumor of the female genital tract in developed countries. And few studies have been reported on the antitumor activity of BSYX. Therefore, this study aimed to investigate the effect of BSYX on endometrial cancer and make an initial discussion of the underlining mechanisms in Ishikawa cells. Methods and Results. Firstly, 60 SPF female nude mice were randomly divided into control group, model group, BSYX group, and positive group. The models of subcutaneous tumor xenograft of nude mice were established by injection of human endometrial carcinoma cell line Ishikawa tumor cell suspension. Compared with model group, BSYX reduced effectively tumor volume and changed pathological feature in mice tumor issue. Meanwhile, proteins from tumor issues were detected by western blot analysis. The protein levels of follicle-stimulating hormone receptor (FSHR), p-Akt/Akt, Gankyrin, and cyclinD1 in the model group were higher than those in control group but the expression in BSYX group was lower than that in the model group. The hypoxia inducible factor alpha (HIF-α) protein level in the model group was lower than those in control group and upregulated in BSYX group. In addition, Ishikawa cells were cultured and then exposed to follicle-stimulating hormone (FSH), LY294002, a highly selective PI3K inhibitor and serum containing BSYX, respectively. LY294002 and BSYX markedly decreased the cancer cell viability and migration ability and increased the apoptosis rate. FSH promoted the cancer cell ability and migration ability. LY294002 and BSYX evidently downregulated the proteins levels of FSHR, p-Akt/Akt, Gankyrin, and cyclinD1 and upregulated the expression of HIF-α protein, and FSH was on the opposite. Conclusions. Taken together, our results showed that the formulation of BSYX had antitumor effect on endometrial cancer in vivo and in vitro and was related with FSH/PI3K/AKT/Gankyrin/HIF-α/cyclinD1 transduction pathway.

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Characterisation of Osteopontin in an In Vitro Model of Embryo Implantation

Cells ◽

10.3390/cells8050432 ◽

2019 ◽

Vol 8 (5) ◽

pp. 432 ◽

Cited By ~ 3

Author(s):

Stéphane C Berneau ◽

Peter T Ruane ◽

Daniel R Brison ◽

Susan J Kimber ◽

Melissa Westwood ◽

...

Keyword(s):

Secretory Pathway ◽

Embryo Implantation ◽

Giant Cells ◽

Ishikawa Cell ◽

Altered Expression ◽

Ishikawa Cells ◽

Cell Layers ◽

Endometrial Epithelium ◽

Vitro Model

At the onset of pregnancy, embryo implantation is initiated by interactions between the endometrial epithelium and the outer trophectoderm cells of the blastocyst. Osteopontin (OPN) is expressed in the endometrium and is implicated in attachment and signalling roles at the embryo–epithelium interface. We have characterised OPN in the human endometrial epithelial Ishikawa cell line using three different monoclonal antibodies, revealing at least nine distinct molecular weight forms and a novel secretory pathway localisation in the apical domain induced by cell organisation into a confluent epithelial layer. Mouse blastocysts co-cultured with Ishikawa cell layers served to model embryo apposition, attachment and initial invasion at implantation. Exogenous OPN attenuated initial, weak embryo attachment to Ishikawa cells but did not affect the attainment of stable attachment. Notably, exogenous OPN inhibited embryonic invasion of the underlying cell layer, and this corresponded with altered expression of transcription factors associated with differentiation from trophectoderm (Gata2) to invasive trophoblast giant cells (Hand1). These data demonstrate the complexity of endometrial OPN forms and suggest that OPN regulates embryonic invasion at implantation by signalling to the trophectoderm.

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HEC-1 Cells: Establishment of an In Vitro Experimental System in Endometrial Carcinoma

Cell and Molecular Biology of Endometrial Carcinoma ◽

10.1007/978-4-431-53981-0_1 ◽

2003 ◽

pp. 3-34 ◽

Cited By ~ 6

Author(s):

Hiroyuki Kuramoto ◽

Mieko Hamano ◽

Manami Imai ◽

Takesi Fujisawa ◽

Yuko Kamata ◽

...

Keyword(s):

Endometrial Carcinoma ◽

Experimental System

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CircATRNL1 promotes epithelial–mesenchymal transition in endometriosis by upregulating Yes-associated protein 1 in vitro

Cell Death and Disease ◽

10.1038/s41419-020-02784-4 ◽

2020 ◽

Vol 11 (7) ◽

Author(s):

Dandan Wang ◽

Yajuan Luo ◽

Guangwei Wang ◽

Qing Yang

Keyword(s):

Epithelial Mesenchymal Transition ◽

Quantitative Polymerase Chain Reaction ◽

Polymerase Chain Reaction Analysis ◽

In Vitro Assays ◽

Migration And Invasion ◽

Circular Rnas ◽

Mesenchymal Transition ◽

Inhibitory Function ◽

Ishikawa Cells

Abstract Endometriosis is a common and benign gynecological disorder but exhibits malignant features. However, the underlying pathogenesis and pathophysiology of endometriosis remain unclear. Circular RNAs have been demonstrated to participate in the occurrence and progression of multiple diseases. This study was aimed to explore the roles of circATRNL1 in endometriosis in vitro. Based on the results of reverse transcription-quantitative polymerase chain reaction analysis, we found significant upregulation of circATRNL1 and Yes-associated protein 1 (YAP1), while downregulation of miR-141-3p and miR-200a-3p in ectopic tissues compared to eutopic tissues. The immunohistochemistry and western blot analysis showed differentially expressed epithelial–mesenchymal transition (EMT) markers between EuEM and EcEM tissues. The in vitro assays indicated that overexpression of circATRNL1 could promote the proliferation, migration, and invasion of Ishikawa cells, and induce EMT process, while circATRNL1 silencing showed the opposite effect. The mechanical investigation indicated that circATRNL1 upregulated YAP1 by sponging miR-141-3p and miR-200a-3p. Gain-of-function assays validated the inhibitory function of miR-141-3p and miR-200a-3p in endometriosis. The results of rescue assays confirmed the function of circATRNL1–miR-141-3p/miR-200a-3p–YAP1 axis on Ishikawa cells. Our findings demonstrate that abnormal upregulation of circATRNL1 regulates cell proliferation and motility and promotes EMT process via the miR-141-3p/miR-200a-3p–YAP1 axis in vitro, which could contribute to the progression of endometriosis.

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