Fig 1.
Expression of EMT markers in prostate cell lines.
(A) Morphology of Prostate Cancer cell lines. (B)The expression of EMT markers (E-cadherin, vimentin, Snail, Cat L, Twist, and Occludin) were examined by western blot analysis. Alpha tubulin was used as the loading control. (C) Zymogram of Cat L activity in various prostate cell lines. (D) Cat L activity was quantified by densitometry for each band on the gel using NIH image J. (n = 3, *** p < 0.001, **p < 0.01, * p < 0.05). Values were normalized to untreated controls and the mean± SEM of data were obtained from three independent replicate experiments. Statistical analysis was done with Student t test.
Fig 2.
mRNA analysis of EMT markers in CA and AA cell lines.
Prostate cell lines were probed for mRNA levels by real-time PCR analysis using primers specific for E-cadherin, Snail, vimentin and Cat L. Results are representative of at least two independent experiments performed in triplicate.
Fig 3.
AA cell lines are highly proliferative and migratory.
CA (LNCaP, C4-2, ARCaP-E, ARCaP-M) and AA (E006AA, E006AA-hT, MDA-PCa-2b) cells were analysed for (A) cell viability using MTS and (B) transwell migration across a collagen-coated boyden chamber. (*** p < 0.001, * p < 0.05). Mean± SEM of data were obtained from three independent replicate experiments. Statistical analysis was done with Student t test.
Fig 4.
TNBC cell lines are more mesenchymal, proliferative and migratory as compared to non-TNBC cell lines.
(A) The morphology of estrogen-receptor (ER)-positive or non-TNBC cell lines (MCF-7, T47-D, MDA-MB-361 and BT-474) and TNBC cell lines (MDA-MB-231, HS-578T, BT-549, MDA-MB-468 and HCC-1806) are shown. (B) TNBC and non-TNBC cells were analyzed for EMT marker expression by western blot analysis. Alpha tubulin was used as the loading control. (C) transwell migration across a collagen-coated boyden chamber was performed. (*** p < 0.001, **** p < 0.0001). Mean± SEM of data were obtained from three independent replicate experiments. Statistical analysis was done with Student t test.
Fig 5.
Snail and Cat L expression in prostate patient tissue.
(A) Immunohistochemical (IHC) analysis was performed using normal and tumor matched prostate adenocarcinoma tissue from Caucasian American (CA) African American (AA) men. For AA: n = 36 normal, n = 38 cancer and for CA: n = 33 normal, n = 45 cancer. Representative images are shown at 20X. (B) IHC for Snail and Cat L was similarly performed for Bahamian (BAH) men normal and tumor-matched tissue (n = 34 normal, n = 22 cancer) and representative images shown at 20X. Inset, 40X mag. Nuclear staining for (C) Snail and (D) Cat L was digitally scored using Image Scanner and verified by a Pathologist and graphed. Points represent nuclear Snail staining intensity of individual CA and AA patients; bars represent the median value for the sample set.
Fig 6.
Snail and Cat L expression in breast patient tissue.
Immunohistochemical (IHC) analysis was performed using normal, ER-positive or luminal A and TNBC breast tissue from Caucasian American (CA), and African American (AA) women with antibody against (A) Snail or (B) Cat L. For AA: normal (n = 24), luminal A (n = 7), TNBC (n = 15). For CA: normal (n = 10), luminal A (n = 14), TNBC (n = 13). Representative images are shown at 10X. Inset, 20X mag. Nuclear staining for (C) Snail and (D) Cat L was digitally scored using Image Scanner and verified by a Pathologist and graphed. Bars represent the median value for the sample set.