mouse op9 preadipocyte cell line  (ATCC)

Journal: Oncology reports

Article Title: Adipocyte culture medium stimulates invasiveness of MDA-MB-231 cell via CCL20 production.

doi: 10.3892/or_00000593

Figure Lengend Snippet: Figure 1. CCL20 production is increased by adipocyte-CM. (A) MDA-MB-231 cells were treated with preadipocyte-CM or adipocyte-CM for 16 h, and supernatants were centrifuged to remove cells. Chemokines in supernatants were analyzed. (B) MDA-MB-231 cells were incubated with preadipocyte- CM or adipocyte-CM for 6 h. Total RNA was isolated and subjected to RT-PCR analysis, and band density was measured. The experiment was performed in triplicate. A representative experiment is shown. (C) MDA-MB-231 cells were incubated with adipocyte-CM for indicated times. CCL20 mRNA expression was upregulated after stimulation with adipocyte-CM. The experiment was performed in triplicate. A representative experiment is shown.

Article Snippet: The MDA-MB-231 human breast cancer cell line and the OP9 mouse preadipocyte cell line were purchased from the American Type Culture Collection (ATCC).

Techniques: Incubation, Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Oncology reports

Article Title: Adipocyte culture medium stimulates invasiveness of MDA-MB-231 cell via CCL20 production.

doi: 10.3892/or_00000593

Figure Lengend Snippet: Figure 2. A protein factor in adipocyte-CM activates NF-κB with a resultant increase in CCL20 mRNA expression in MDA-MB-231 cells. (A) MDA- MB-231 cells were treated with preadipocyte-CM, adipocyte-CM, or pro- teinase K-treated adipocyte CM, for 6 h. Total RNA was isolated and subjected to RT-PCR analysis. (B) MDA-MB-231 cells were pretreated with 20 μM SP600125, 20 μM SB203580, 20 μM PD98059, or 1.5 μM parthenolide (PTN), 1 h before treatment with adipocyte-CM, and incubated for 6 h. Total RNA was isolated and subjected to RT-PCR analysis.

Article Snippet: The MDA-MB-231 human breast cancer cell line and the OP9 mouse preadipocyte cell line were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Isolation, Reverse Transcription Polymerase Chain Reaction, Incubation

Journal: Oncology reports

Article Title: Adipocyte culture medium stimulates invasiveness of MDA-MB-231 cell via CCL20 production.

doi: 10.3892/or_00000593

Figure Lengend Snippet: Figure 3. The TNF-· gene is not more highly expressed in adipocytes than in preadipocytes. (A) MDA-MB-231 cells were treated with TNF-· (10 ng/ml) or IL-1ß (15 ng/ml) for 6 h. The levels of CCL20 mRNA expression were examined by RT-PCR. (B) OP9 preadipocytes were permitted to differentiate into adipocytes. Cells were harvested at the indicated days during differentiation. The levels of TNF-· and IL-1ß gene expression were examined using RT-PCR analysis.

Article Snippet: The MDA-MB-231 human breast cancer cell line and the OP9 mouse preadipocyte cell line were purchased from the American Type Culture Collection (ATCC).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Gene Expression

Journal: Oncology reports

Article Title: Adipocyte culture medium stimulates invasiveness of MDA-MB-231 cell via CCL20 production.

doi: 10.3892/or_00000593

Figure Lengend Snippet: Figure 4. Anti-TNF-· neutralizing antibody blocks adipocyte-CM-induced CCL20 production in MDA-MB-231 cells. (A) MDA-MB-231 cells were treated with preadipocyte-CM, adipocyte-CM, or proteinase K-treated adipocyte-CM, for 6 h. The levels of TNF-· and IL-1ß gene expression were examined by RT- PCR. (B) MDA-MB-231 cells were treated with adipocyte-CM, and total RNA was isolated at the indicated times. The levels of TNF-· gene expression were examined by RT-PCR. (C) MDA-MB-231 cells were treated with adipocyte-CM preincubated with various concentrations of anti-TNF-· neutralizing antibody for 1 h. The level of CCL20 expression was examined by RT-PCR 6 h after incubation with adipocyte-CM containing anti-TNF-· neutralizing antibody. (D) Preadipocytes were treated with TNF-· siRNA and GFP siRNA as a control for 24 h and then differentiated into adipocytes. MDA-MB-231 cells were treated with TNF-·-deficient preadipocyte-CM and TNF-·-deficient adipocyte-CM for 6 h. The level of CCL20 expression was examined by RT-PCR. (E) MDA-MB-231 cells were treated with TNF-· receptor siRNA or GFP siRNA as a control for 24 h and then preadipocyte-CM or adipocyte- CM was treated for 6 h. The level of CCL20, TNF-·, and TNF-· receptor 1 expression was examined by RT-PCR. (F) MDA-MB-231 cells were treated with TNF-· receptor siRNA or GFP siRNA as a control for 24 h and then preadipocyte-CM or adipocyte-CM was treated for 30 min. Lysates were used to determine the level of phospho-IKK·/ß and phospho-IκB· using Western blot analysis.

Article Snippet: The MDA-MB-231 human breast cancer cell line and the OP9 mouse preadipocyte cell line were purchased from the American Type Culture Collection (ATCC).

Techniques: Gene Expression, Reverse Transcription Polymerase Chain Reaction, Isolation, Expressing, Incubation, Control, Western Blot