Role of peptidylarginine deiminase 2 (PAD2) in mammary carcinoma cell migration

Abstract We investigated the role of autocrine production of human (h) GH in the attachment and spreading of mammary carcinoma cells in vitro. We used a previously described model system for the study of the autocrine/paracrine role of GH in which the hGH gene (MCF-hGH) or a translation-deficient hGH gene (MCF-MUT) was stably transfected into MCF-7 cells. No differences in attachment to a collagen matrix between MCF-hGH and MCF-MUT cells were observed in either serum-free medium (SFM) or medium containing exogenous hGH, 5% serum, or 10% serum. In contrast, MCF-hGH cells spread more rapidly on a collagen matrix than did MCF-MUT cells. Exogenous hGH and 10% serum interacted with autocrine production of hGH in an additive manner to increase cell spreading. MCF-hGH cells formed filipodia and stress fibers earlier than MCF-MUT cells during the process of cell spreading and possessed marked differences in morphology after spreading. MCF-MUT cells displayed uniform and symmetrical formation of stress fibers, whereas MCF-hGH cells displayed irregular and elongated stress fiber formation. The level of cytoplasmic phosphotyrosine was increased in MCF-hGH compared with MCF-MUT cells during spreading and displayed colocalization with Janus kinase 2 (JAK2). Basal JAK2 tyrosine phosphorylation was increased, and it increased further on spreading in MCF-hGH cells compared with MCF-MUT cells. Transient transfection of JAK2 complementary DNA resulted in interaction with autocrine hGH to increase the rate of cell spreading in MCF-hGH cells compared with MCF-MUT cells. Treatment with a selective JAK2 tyrosine kinase inhibitor (AG 490) reduced the rate of MCF-hGH cell spreading to the rate of MCF-MUT cell spreading. Thus, we conclude that autocrine production of hGH enhances the rate of mammary carcinoma cell spreading in a JAK2-dependent manner.

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Aquaporin-1 Activity of Plasma Membrane Affects MCF-7 Mammary Carcinoma Cell Migration

2009 3rd International Conference on Bioinformatics and Biomedical Engineering ◽

10.1109/icbbe.2009.5162404 ◽

2009 ◽

Author(s):

Yong Jiang ◽

Xin Zhang ◽

Rui Su ◽

Nan Wang

Keyword(s):

Plasma Membrane ◽

Cell Migration ◽

Mammary Carcinoma ◽

Carcinoma Cell ◽

Mammary Carcinoma Cell ◽

Aquaporin 1 ◽

Mcf 7

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In Vitro Effects of Polar Materials from Thermally Abused Frying Oil on Mammary Carcinoma Cell Migration (P05-004-19)

Current Developments in Nutrition ◽

10.1093/cdn/nzz030.p05-004-19 ◽

2019 ◽

Vol 3 (Supplement_1) ◽

Author(s):

Kaitlyn Joyce ◽

Cheng Chen ◽

Jennifer Hughes ◽

Ashley Oyirifi ◽

William Helferich

Keyword(s):

Cell Migration ◽

Mammary Carcinoma ◽

Wound Closure ◽

Carcinoma Cell ◽

Polar Compounds ◽

Frying Oil ◽

Deep Frying ◽

Mammary Carcinoma Cell ◽

Polar Material

Abstract Objectives Widely consumed foods such as chicken, fish, and potatoes are regularly prepared by deep frying. The frying process involves temperatures exceeding 180 degrees Celsius and repeated frying cycles that result in thermally induced chemical changes of the oil's lipid structures. One such chemical change is an accumulation of polar compounds, including secondary lipid oxidation products, which are associated with several disease pathologies. Many European countries adhere to strict limits of less than 30% polar compounds within recycled fryer oils. There are no such regulations in the United States. Using a murine model of late-stage breast cancer (BC), we previously demonstrated an increased metastatic burden in mice consuming a diet of thermally abused frying oil (TAFO) compared with mice consuming fresh vegetable oil. To further understand this observation, we assessed 1) the amount of polar material in oil recycled for 300 minutes of deep frying, and 2) the effect the fractionated polar material from TAFO has on in vitro migration of 4T1 murine cancer cells. Methods We used silica column chromatography to separate the TAFO into polar and non-polar fractions. The polar fraction of TAFO (TAFO-PF) was retained from oil used to fry fish nuggets for a duration of 300 minutes. In vitro wound healing migration assays were conducted in the presence or absence of TAFO-PF in a concentration dependent manner. We assessed the wound closure rates (motility) of the highly metastatic 4T1 murine mammary carcinoma cell line. Live images were captured every hour for 24 hours to measure cell migration using brightfield microscopy. Results We found that after 300 minutes of frying, oil contained 74 ± 7.8 μg/mL of polar compound. 4T1 cells incubated over a period of 24 hours with diluted TAFO-PF achieved faster wound closure rates compared with cells incubated in growth media alone. Conclusions Our results suggest that TAFO-PF increases motility as an indicator of the metastatic potential of BC cells. Ongoing work is being focused on conducting in vitro invasion assays on both 4T1 and human BC MDA-MB-231 cell lines in order to further understand the potential mechanisms and effects TAFO-PF has on cancer metastatic progression. Funding Sources NIEHS Training (T32) Grant ES007326 Fellowship to JRH and ABO; UIUC Campus Research Board Beckman Grant and UIUC Hatch 1011659_ILLU-698-357.

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