Epigenetic Inactivation of EFEMP1 Is Associated with Tumor Suppressive Function in Endometrial Carcinoma

Yang, Tingting; Qiu, Haifeng; Bao, Wei; Li, Bilan; Lu, Cong; Du, Guiqiang; Luo, Xin; Wang, Lihua; Wan, Xiaoping

doi:10.1371/journal.pone.0067458

Cited by 34 publications

(41 citation statements)

References 36 publications

(39 reference statements)

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“…Sadr-Nabavi et al determined a similar outcome in sporadic breast cancer using RNA microarray expression analyses (10). Of note, our findings also correlate with demonstrations of EFEMP1 downregulation reported in instances of endometrial cancer by Yang et al (18). According to this study, the decrease in EFEMP1 expression is mainly regulated by promoter hypermethylation, a finding consistent with our preliminary results that suggest differential methylation patterns in leiomyomata.…”

Section: Discussionsupporting

confidence: 92%

Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 expression and regulation in uterine leiomyoma

Marsh

Chibber

et al. 2016

Fertility and Sterility

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Objective To determine the presence, differential expression and regulation of epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) in uterine leiomyomas. Design Laboratory in vivo and in vitro study using human leiomyoma and myometrial tissue and primary cells. Setting Academic medical center Samples Leiomyoma and myometrial tissue samples and cultured cells Intervention(s) 5-Aza-2′ deoxycytidine (5-Aza-dC) treatment. Main Outcome Measure Fold change difference between EFEMP1 and fibulin-3 expression in leiomyoma tissue and cells compared to matched myometrial samples, and fold-change difference in EFEMP1 expression with 5-Aza-dC treatment. Results In vivo, EFEMP1 expression was 3.19 fold higher in myometrial tissue versus leiomyoma tissue. EFEMP1 expression in vitro was 5.03 fold higher in myometrial cells versus leiomyoma cells. Western blot and IHC staining of tissue and cells confirmed similar findings in protein expression. Treatment of leiomyoma cells with 5-Aza-dC resulted in increased expression of EFEMP1 in vitro. Conclusion The EFEMP1 gene and its protein product, fibulin-3, are both significantly downregulated in leiomyoma as compared to myometrium when studied both in vivo and in vitro. The increase in EFEMP1 expression in leiomyoma cells with the 5-Aza-dC treatment suggest that differential methylation is, in part, responsible for the differences seen in gene expression.

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Section: Discussionsupporting

confidence: 92%

Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 expression and regulation in uterine leiomyoma

Marsh

Chibber

et al. 2016

Fertility and Sterility

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“…EFEMP1 abrogated angiogenic activities and sprouting in MB114 cells by decreasing the expression of MMP-2 and MMP-3 (13). EFEMP1 is associated with decreased MMP-2 and MMP-7 levels in lung cancer (14) and MMP-2 and MMP-9 levels in endometrial carcinoma (15), which are somewhat similar to our data. We found that downregulation of EFEMP1 increased the expression of MMP2 and MMP9, and MMP2 and MMP9 level in clinical samples showed the same tendency: the lower the EFEMP1 the higher the MMP2 and MMP9 expression was.…”

Section: Discussionsupporting

confidence: 90%

EFEMP1 inhibits migration of hepatocellular carcinoma by regulating MMP2 and MMP9 via ERK1/2 activity

et al. 2016

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The role of epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) inhibiting migration in hepatocellular carcinoma (HCC) remains unknown. Expression of EFEMP1 in HCC cell lines were quantified by western blotting and real-time PCR. The role of EFEMP1 in HCC cell migration was explored in vitro via siRNA and adding purified EFEMP1 protein. The associated molecule expression was detected by western blotting after downregulation of EFEMP1 and also tested by immunohistochemistry. Eight pairs of HCC non-HCC liver samples and 215 HCC samples were subjected to immunohistochemistry. EFEMP1 was highly expressed in 7,721 and HepG2 HCC cell lines while HuH7 HCC cell line expressed the lowest level of EFEMP1 compared with the others. Downregulating EFEMP1 by siRNA markedly increased the migration ability of HCC cells while adding purified EFEMP1 protein inhibited HCC cell migration. Downregulation of EFEMP1 increased the expression of ERK1/2, MMP2 and MMP9. Furthermore, U0126 (a highly selective and potent inhibitor of pERK1/2) could abrogate the migration ability enhanced by siRNA. Accordingly, MMP2 and MMP9 were inversely expressed with EFEMP1 expression by immunohistochemistry. EFEMP1 downregulated in HCC tissues, and lower EFEMP1 expression was significantly associated with HCC patients with ascites (P=0.050), vascular invasion (P=0.044), poorer differentiation (P=0.002) and higher clinical stage (P=0.003).

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“…The primers for amplification of IL‐6, total aromatase mRNA and aromatase promoter I.3, I.4, I.7 and II mRNA are shown in the Supporting Information. RNA extraction, real‐time RT‐PCR and western blot were performed as described previously . IHC was performed as described previously and evaluation of aromatase and IL‐6 staining was performed based on staining intensity proportion scoring systems used for breast carcinoma tissues (See Supporting Information).…”

Section: Coculture Experimentsmentioning

confidence: 99%

Activation of a positive feedback loop involving IL-6 and aromatase promotes intratumoral 17β-estradiol biosynthesis in endometrial carcinoma microenvironment

et al. 2014

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Tumor-stroma interactions contribute greatly to intratumoral estrogen biosynthesis in endometrial carcinoma, but the mechanisms involved remain largely unknown. Previous study demonstrated that intratumoral aromatase upregulation in stromal cells participated in this process, but the specific aromatase-regulators have not been reported. In the present study, we found that aromatase expression in intratumoral stroma, but not in tumor epithelium, correlated positively with interleukin 6 (IL-6) expression in cancer epithelial cells by immunohistochemistry, which was confirmed using laser capture microdissection/real-time reverse transcription-PCR. With stimulation by exogenous IL-6, aromarase expression was increased in stromal cells not but not in cancer cells. Aromatase mRNA levels in endometrial cancer cells were not influenced by cocultivation with intratumoral stromal cells. When cocultured with 17b-estradiol (E 2 )-treated cancer cells, aromatase mRNA in stromal cells was significantly elevated and increased IL-6 protein levels were detected in E 2 -treated culture medium. Next, we demonstrated that E 2 -induced IL-6 production was through cooperation between estrogen receptor a and nuclear factor-kappa B. Furthermore, an IL-6 receptor blocking antibody could attenuate the upregulation of aromatase expression in stromal cells and the E 2 concentration in coculture systems of cancer and stromal cells. The results were confirmed by an orthotopic nude endometrial carcinoma model in vivo. These studies elucidated the activation of a positive feedback loop, that is, IL-6 stimulated by E 2 in endometrial cancer cells induced aromatase expression in stromal cells, promoting enhanced intratumoral E 2 synthesis. Blocking of this tumor-stroma interaction may be a therapeutic strategy to overcome in situ estrogen biosynthesis in endometrial carcinoma.Endometrial carcinoma is the most common gynecologic malignancy, with an estimated 47,130 diagnosed cases and 8,010 deaths in 2012 in the United States. 1 Existing evidence indicates that unopposed estrogens contribute to the tumorigenesis and promotion of endometrial carcinoma. 2 The conversion of androstenedione and testosterone into estrogen is catalyzed by the cytochrome P450 aromatase enzyme. 3 Although postmenopausal women have low levels of circulating plasma estrogens, the intratumoral production of estrogens can lead to higher estrogen levels in tumors. 4 Several studies have indicated that, in normal and abnormal human endometrium, aromatase expression and activity are associated with malignancy and only aromatase expression in stromal cells, but not in epithelial cells, correlates positively with poor survival. 5,6 Additionally, an indirect coculture model confirmed that tumor-stroma communication contributed to upregulation of aromatase activity. 7 Therapeutic strategies that focus on tumor-stroma interactions require the identification of molecular targets that regulate intercellular interactions. To the best of our knowledge, however, no exact stimulators have...

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Epigenetic Inactivation of EFEMP1 Is Associated with Tumor Suppressive Function in Endometrial Carcinoma

Cited by 34 publications

References 36 publications

Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 expression and regulation in uterine leiomyoma

Epidermal growth factor–containing fibulin-like extracellular matrix protein 1 expression and regulation in uterine leiomyoma

EFEMP1 inhibits migration of hepatocellular carcinoma by regulating MMP2 and MMP9 via ERK1/2 activity

Activation of a positive feedback loop involving IL-6 and aromatase promotes intratumoral 17β-estradiol biosynthesis in endometrial carcinoma microenvironment

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