Journal: BMC Cancer

Article Title: Conjugated linoleic acid induces apoptosis through estrogen receptor alpha in human breast tissue

doi: 10.1186/1471-2407-8-208

Figure Lengend Snippet: Effects of CLA on ERα(+) human breast cells. (A) MTS assay, (B) Apo-ONE ® homogeneous caspase-3/7 assay and (C) Hoechst staining for 3 days of treatment with CLA, Tam and E 2 alone or combined with each other in MCF-7 cells. CLA stands for 40 μM t 10, c 12-CLA; Tam stands for 1 μM 4-Hydroxytamoxifen; E 2 stands for 10 nM 17β-estradiol. CLA exerts combinative effects with tamoxifen in MCF-7 cells. (D) Western blot analysis of basal ERα protein expression in MCF-7 and primary cultured ERα positive normal human breast epithelial cells (ERα(+) NEC). Equal amounts of isolated protein from both cell extracts were subjected to immunoblot with anti-ERα antibodies. β-actin was used as loading control. Histogram: Apo-ONE ® homogeneous caspase-3/7 assay for the effects of treatment on apoptosis in MCF-7 and ERα(+) NEC for 3 days of treatment. (E) and (F) showed the effects of CLA and E 2 on proliferation (MTS assay) and apoptosis (Hoechst staining) in ERα(+) NEC, respectively. 80 μM t 10, c 12-CLA; E 2 stands for 10 nM 17β-estradiol. Bars represent mean ± SD, n = 3. *p < 0.05.

Article Snippet: t 10, c 12-CLA (> 98% pure), the CLA isomer used in this study, was purchased from Matreya, Inc. (Greenland, NH) and a CLA stock solution was prepared as described previously [ ].

Techniques: MTS Assay, Staining, Western Blot, Expressing, Cell Culture, Isolation

Journal: BMC Cancer

Article Title: Conjugated linoleic acid induces apoptosis through estrogen receptor alpha in human breast tissue

doi: 10.1186/1471-2407-8-208

Figure Lengend Snippet: Effects of CLA, Tam and E 2 on cell proliferation and apoptosis of ERα negative human breast epithelial cells. (A) and histogram in (C) are MTS assay for the effects of CLA, Tam and E 2 for 3 days of treatment on cell proliferation of MDA-MB-231 and ERα negative normal human breast epithelial cells (ERα(-) NEC), respectively. (C) Western blot analysis of basal ERα protein expression in MCF-7 and primary cultured ERα negative normal human breast epithelial cells (ERα(-) NEC). Equal amounts of isolated protein from both cell extracts were subjected to immunoblot with anti-ERα antibodies. β-actin was used as loading control. Hoechst staining of (B) MDA-MB-231 and (E) ERα(-) NEC, and (D) Apo-ONE ® homogeneous caspase-3/7 assay of ERα(-) NEC after 3 days of CLA, Tam and E 2 treatment. CLA stands for 40 μM t 10, c 12-CLA. Tam stands for 1 μM 4-Hydroxytamoxifen; E 2 stands for 10 nM 17β-estradiol. *p < 0.05 versus control. Docetaxel, the commonly used anti-cancer drug, was used as the positive control in the assay of cell proliferation and apoptotic activity.

Article Snippet: t 10, c 12-CLA (> 98% pure), the CLA isomer used in this study, was purchased from Matreya, Inc. (Greenland, NH) and a CLA stock solution was prepared as described previously [ ].

Techniques: MTS Assay, Western Blot, Expressing, Cell Culture, Isolation, Staining, Positive Control, Activity Assay

Journal: BMC Cancer

Article Title: Conjugated linoleic acid induces apoptosis through estrogen receptor alpha in human breast tissue

doi: 10.1186/1471-2407-8-208

Figure Lengend Snippet: CLA induces apoptosis in ERα positive human breast cancer cell line. (A) ERα protein expression was examined by western blot of whole cell lysate isolated from MCF-7, MDA-MB-231 (parental), and MDA-MB-231-ERα (ERα transfected MDA-MB-231). Equal amounts of isolated protein from cell extracts of all three cell lines were subjected to immunoblot with anti-ERα antibodies. β-actin was used as loading control. Effects of CLA on apoptosis of MCF-7, MDA-MB-231, and MDA-MB-231-ERα were determined by Hoechst staining (B) and Apo-ONE ® homogeneous caspase-3/7 assay (C). Bars represent mean ± SD, n = 3. *p < 0.05 stands for control versus 40 μM t 10, c 12-CLA.

Article Snippet: t 10, c 12-CLA (> 98% pure), the CLA isomer used in this study, was purchased from Matreya, Inc. (Greenland, NH) and a CLA stock solution was prepared as described previously [ ].

Techniques: Expressing, Western Blot, Isolation, Transfection, Staining

Journal: BMC Cancer

Article Title: Conjugated linoleic acid induces apoptosis through estrogen receptor alpha in human breast tissue

doi: 10.1186/1471-2407-8-208

Figure Lengend Snippet: CLA decreases E 2 stimulated Bcl-2 protein expression in ERα(+) breast cells. (A) Immunohistochemistry of Bcl-2 staining (brown to dark) in ERα(+) normal human breast tissue for 3 days of treatment. (B) Western blot analysis of basal ERα protein expression in MCF-7 and ERα(+) NEC, and the effects of CLA and E 2 on Bcl-2 protein expression in ERα(+) NEC for 3 days of treatment. β-actin was used as loading control. (C) Western blot analysis of basal ERα protein expression in MCF-7 and ERα positive human breast cancer epithelial cells (ERα(+) CAEC). And Bcl-2 protein expression in MCF-7 and ERα(+) CAEC was detected by western blot analysis after 3 days of treatment. CLA exerts combinative effects with Tam in decreasing E 2 stimulated Bcl-2 protein expression in ERα(+) CAEC. CLA stands for 40 μM t 10, c 12-CLA; E 2 stands for 10 nM 17β-estradiol.

Article Snippet: t 10, c 12-CLA (> 98% pure), the CLA isomer used in this study, was purchased from Matreya, Inc. (Greenland, NH) and a CLA stock solution was prepared as described previously [ ].

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot

Journal: BMC Cancer

Article Title: Conjugated linoleic acid induces apoptosis through estrogen receptor alpha in human breast tissue

doi: 10.1186/1471-2407-8-208

Figure Lengend Snippet: CLA induced apoptosis of breast cancer epithelial cells is associated with estrogen receptor α (ERα) in co-culture. MCF-7 (A), MDA-MB-231 (B), ERα transfected MDA-MB-231, and MDA-MB-231-ERα (C), were cultured alone or co-cultured with normal human breast stromal cells (NSC), or co-cultured with cancerous human breast stromal cells (CASC). Cells were treated by t 10, c 12-CLA for 7 days and cell apoptosis of MCF-7 (A), MDA-MB-231 (B) or MDA-MB-231-ERα (C) was detected from each group separately by caspase-3/7 activity assay. (D) Bcl-2 protein expression of MCF-7, MDA-MB-231 or MDA-MB-231-ERα was determined by western blot analysis. CLA stands for 40 μM t 10, c 12-CLA. Bars represent mean ± SD, n = 3. *p < 0.05 stands for t 10, c 12-CLA group comparison of caspase-3/7 activity; +p < 0.05 for control versus t 10, c 12-CLA.

Article Snippet: t 10, c 12-CLA (> 98% pure), the CLA isomer used in this study, was purchased from Matreya, Inc. (Greenland, NH) and a CLA stock solution was prepared as described previously [ ].

Techniques: Co-Culture Assay, Transfection, Cell Culture, Activity Assay, Expressing, Western Blot