Extracellular Matrix Composition and Hypoxia Regulate the Expression of HLA-G and Integrins in a Human Trophoblast Cell Line1

Kilburn, Brian A.; Wang, Jun; Duniec-Dmuchkowski, Zophia M.; Leach, Richard E.; Romero, Roberto; Armant, D. Randall

doi:10.1095/biolreprod62.3.739

Cited by 181 publications

(140 citation statements)

References 28 publications

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“…Altered expressions of HLA-I and HLA-E molecules by parental and RCC-pE cells, and the results of blocking experiments, suggest that VHL is responsible for the susceptibility of these cells to NK-mediated lysis. The previously reported expression of HLA-G by RCC6 (Ibrahim et al, 2001) is decreased by transfection with the wild-type VHL gene, in agreement with the link between HIF-1 stabilization and HLA-G expression by melanoma cells (Mouillot et al, 2007) and trophoblast (Kilburn et al, 2000). Transfection with the wild-type VHL had no effect on the expression of ligands for NKG2D and DNAM-1 activating NK receptors.…”

Section: Discussionsupporting

confidence: 76%

Mutations of the von Hippel–Lindau gene confer increased susceptibility to natural killer cells of clear-cell renal cell carcinoma

et al. 2011

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The tumor suppressor gene von Hippel-Lindau (VHL) is involved in the development of sporadic clear-cell renal cell carcinoma (RCC). VHL interferes with angiogenesis and also controls cell adhesion and invasion. Therapies that target VHL-controlled genes are currently being evaluated in RCC patients. RCC is a immunogenic tumor and treatment with interleukin-2 (IL2) or interferon (IFN)-a results in regression in some patients. We used two renal tumor cell lines (RCC6 and RCC4) carrying VHL loss-of-function mutations to investigate the role of mutant VHL in susceptibility to natural killer (NK) cellmediated lysis. The RCC6 and RCC4 cell lines were transfected with the wild-type gene to restore the function of VHL. The presence of the gene in RCC cells downregulated hypoxia-inducible factor (HIF)-1a and subsequently decreased vascular endothelial growth factor (VEGF) production. Relative to control transfectants and parental cells, pVHL-transfected cell lines activated resting and IL2-activated NK cells less strongly, as assessed by IFNc secretion, NK degranulation and cell lysis. NKG2A, a human leukocyte antigen (HLA)-Ispecific inhibitory NK receptor, controls the lysis of tumor targets. We show that HLA-I expression in RCC-pVHL cells is stronger than that in parental and controls cells, although the expression of activating receptor NK ligands remains unchanged. Blocking NKG2A/HLA-I interactions substantially increased lysis of RCC-pVHL, but had little effect on the lysis of VHL-mutated RCC cell lines. In addition, in response to IFNa, the exponential growth of RCC-pVHL was inhibited more than that of RCC-pE cells, indicating that VHL mutations may be involved in IFNa resistance. These results indicate that a decreased expression of HLA-I molecules in mutated VHL renal tumor cells sensitizes them to NK-mediated lysis. These results suggest that combined immunotherapy with antiangiogenic drugs may be beneficial for patients with mutated VHL.

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Section: Discussionsupporting

confidence: 76%

Mutations of the von Hippel–Lindau gene confer increased susceptibility to natural killer cells of clear-cell renal cell carcinoma

et al. 2011

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“…This possibility is supported by recent report of Malmberg et al [12] that HLA-G protein was detected only in freshly isolated ovarian carcinoma cells, but not in cells after long-term culture. The mechanism(s) by which HLA-G expression is modulated in melanoma cells have been investigated, such as stress [13], hypoxia and IFNs [14,15], leukemia inhibitory factor and glucocorticoids [16,17]. Others such as epigenetic factors have also been explored in melanoma cells.…”

Section: Introductionmentioning

confidence: 99%

Induction of HLA-G expression in a melanoma cell line OCM-1A following the treatment with 5-aza-2′-deoxycytidine

et al. 2005

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The non-classical HLA class I antigen HLA-G is an immune modulator which inhibits the functions of T cells, NK cells, and the Dendritic cells (DC). As a result, HLA-G expression in malignant cells may provide them with a mechanism to escape the immune surveillance. In melanoma, HLA-G antigen expression has been found in 30% of surgically removed lesions but in less than 1% of established cell lines. One possible mechanism underlying the differential HLA-G expression in vivo and in vitro is that the HLA-G gene is epigenetically repressed in melanoma cells in vitro. To test this hypothesis, we treated the HLA-G negative melanoma cell line OCM-1A with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-AC) and analyzed whether HLA-G expression can be restored. Our data strongly suggest that HLA-G is silenced as a result of CpG hypermethylation within a 5' regulatory region encompassing 220 bp upstream of the start codon. After treatment, HLA-G mRNA expression was dramatically increased. Western blot and flow cytometry showed that HLA-G protein was induced. Interestingly, HLA-G cell surface expression on the 5-AC treated OCM-1A cells is much less than that on the HLA-G positive JEG-3 cells while a similar amount of total HLA-G was observed. Possible mechanisms for the difference were analyzed in the study such as cell cold-treatment, peptide loading and antigen processing machinery components (APM) as well as β 2 microglobulin (β 2 -m) expression. Data revealed that the APM component calreticulin might be involved in the lower HLA-G surface expression on OCM-1A cells. Taken together, our results indicated that DNA methylation is an important epigenetic mechanism by which HLA-G antigen expression is modulated in melanoma cells in vitro. Furthermore, to the first time, we hypothesized that the deficiency of calreticulin might be involved in the low HLA-G surface expression on the 5-AC treated OCM-1A cells.

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“…HTR-8/SVneo cell proliferation increases at 2% O 2 (12,13) as in primary cultured cytotrophoblast and villous explants under similar O 2 conditions (30,31). The inhibition of HBEGF secretion or its binding to HER 1 and HER 4 during exposure to 2% O 2 did not prevent the increase in cell proliferation (12).…”

Section: Commentmentioning

confidence: 96%

“…HTR-8/SVneo cytotrophoblast cells were plated and grown in serum containing medium, which was replaced with serum-free medium 24 hours prior to all experiments, as previously described (13). The cell line was routinely phenotyped to assure cytokeratin-7 expression, hCG secretion, invasion of Matrigel™ basement membrane and HLA-G expression by invading cells.…”

Section: Cell Culture and O 2 Regulationmentioning

confidence: 99%

“…This cell line displays several characteristic features of first trimester cytotrophoblast, including expression of HLA-G, integrin switching from α6β4 to α1β1 during basement membrane invasion, and increased proliferation under physiologic hypoxic conditions (13). We previously demonstrated that HBEGF upregulation is critical for the survival of human cytotrophoblast cells during exposure to low concentrations of O 2 (12), such as those encountered in utero during the first trimester of gestation (15).…”

Section: Commentmentioning

confidence: 99%

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Diminished survival of human cytotrophoblast cells exposed to hypoxia/reoxygenation injury and associated reduction of heparin-binding epidermal growth factor-like growth factor

Leach

Kilburn

Petkova

et al. 2008

American Journal of Obstetrics and Gynecology

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Objective-The anti-apoptotic action of HBEGF and its regulation by O 2 constitutes a key factor for trophoblast survival. The hypothesis that cytotrophoblast survival is compromised by exposure to hypoxia-reoxygenation (H/R) injury, which may contribute to preeclampsia and some missed abortions, prompted us to investigate HBEGF regulation and its role as a survival factor during H/ R in cytotrophoblast cells Study Design-A transformed human first trimester cytotrophoblast cell line HTR-8/SVneo was exposed to H/R (2% O 2 followed by 20% O 2 ) and assessed for HBEGF expression and cell death.Results-Cellular HBEGF declined significantly within 30 minutes of reoxygenation after culture at 2% O 2 . H/R significantly reduced proliferation and increased cell death when compared to trophoblast cells cultured continuously at 2% or 20% O 2 . Restoration of cell survival also was achieved by adding recombinant HBEGF during reoxygenation. HBEGF inhibited apoptosis through its binding to either HER1 or HER4, its cognate receptors.Conclusion-These results provide evidence that cytotrophoblast exposure to H/R induces apoptosis and decreased cell proliferation. HBEGF accumulation is diminished under these conditions, while restoration of HBEGF signaling improves trophoblast survival.

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Extracellular Matrix Composition and Hypoxia Regulate the Expression of HLA-G and Integrins in a Human Trophoblast Cell Line1

Cited by 181 publications

References 28 publications

Mutations of the von Hippel–Lindau gene confer increased susceptibility to natural killer cells of clear-cell renal cell carcinoma

Mutations of the von Hippel–Lindau gene confer increased susceptibility to natural killer cells of clear-cell renal cell carcinoma

Induction of HLA-G expression in a melanoma cell line OCM-1A following the treatment with 5-aza-2′-deoxycytidine

Diminished survival of human cytotrophoblast cells exposed to hypoxia/reoxygenation injury and associated reduction of heparin-binding epidermal growth factor-like growth factor

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