Ethanol-induced in vitro invasion of breast cancer cells: the contribution of MMP-2 by fibroblasts.

Ethanol-induced in vitro invasion of breast cancer cells: the contribution of MMP-2 by fibroblasts.

International journal of cancer (2004-09-24)

Moe Moe Aye, Cuiling Ma, Hong Lin, Kimberly A Bower, Richard C Wiggins, Jia Luo

ABSTRACT

Ethanol is a tumor promoter and may promote metastasis of breast cancer. However, the underlying cellular/molecular mechanisms remain unknown. Overexpression and high activity of matrix metalloproteinase-2 (MMP-2) are frequently associated with metastatic breast cancers and serve as a prognostic indicator of clinical outcome. MMP-2 is predominantly expressed in stromal fibroblasts and plays a pivotal role in regulating the invasive behavior of breast tumor cells. We hypothesized that ethanol may enhance the invasion of breast tumor cells by modulating the activity of fibroblastic MMP-2. With in vitro models (HS68 and CCD1056SK human fibroblasts), we showed that ethanol at physiologically relevant concentrations (50-200 mg/dl) activated MMP-2; conversely, at a higher concentration (400 mg/dl), it inhibited the MMP-2 activity. Consistently, conditioned medium collected from ethanol (50-200 mg/dl)-exposed fibroblasts markedly enhanced the invasive potential of breast cancer cells and mammary epithelial cells overexpressing ErbB2/HER2 (BT474, SKBR-3 and HB2(ErbB2) cells) but had little effect on cells with low ErbB2 levels (BT20 and HB2 cells). In contrast, conditioned medium obtained from ethanol (400 mg/dl)-treated fibroblasts inhibited cell invasion. Selective inhibitors of MMP-2 (SB-3CT and OA-Hy) eliminated ethanol-stimulated invasion, indicating that the effect of ethanol was mediated by MMP-2. Ethanol activated conventional PKCs and JNKs in fibroblasts; inhibitors of PKC (Go6850 and Go6976) and JNKs (SP600125) significantly inhibited ethanol-mediated MMP-2 activation as well as cell invasion, indicating that PKCs and JNKs play a role in ethanol-induced MMP-2 activation and cell invasion in vitro. Thus, ethanol-promoted breast cancer cell invasion may be mediated by the modulation of fibroblastic MMP-2.

MATERIALS

Product Number

Brand

Product Description

E-040

Supelco

Ethanol-10, 10 mg/dL in H₂O, pack of 10 × 1.2 mL ampules, certified reference material, Cerilliant^®

E-041

Supelco

Ethanol-150, 150 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-032

Supelco

Ethanol-200 (10 ampules/kit), 200 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-033

Supelco

Ethanol-300, 300 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-045

Supelco

Ethanol-40, 40 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-030

Supelco

Ethanol-80, 80 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-029

Supelco

Ethanol-50, 50 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-031

Supelco

Ethanol-100 (10 ampules/kit), 100 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

PHR1070

Supelco

Dehydrated Alcohol, Pharmaceutical Secondary Standard; Certified Reference Material

277649

Sigma-Aldrich

Reagent Alcohol, anhydrous, ≤0.003% water

E-053

Supelco

Ethanol-500, 500 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-056

Supelco

Ethanol-20 (10 ampules/kit), 20 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

48075

Supelco

Ethanol solution, certified reference material, 2000 μg/mL in methanol

E-034

Supelco

Ethanol Calibration Kit, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-035

Supelco

Ethanol-25, 25 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®

E-036

Supelco

Ethanol-400 (10 ampules/kit), 400 mg/dL in H₂O, ampule of 10 × 1.2 mL, certified reference material, Cerilliant^®