Figure 1.
Experimental scheme in using HPCa samples. See Text for description.
Table 1.
HPCa xenotransplantation using tumor pieces in immunodeficient micea.
Table 2.
Freshly purified and unsorted HPCa cells injected in NSG micea.
Table 3.
HPCa cells mixed with Hs5 cells initiate serially transplantable tumors in NSG mice*.
Figure 2.
IHC analysis of HPCa57 patient sample (GS7) and its xenograft tumor.
(A) Staining of HE, AR, PSA, CK8, CK5, p63 and Racamase in HPCa57 patient sample. (B) HPCa57P1 xenograft tumor, derived from coinjection with 100,000 Hs5 cells, was used to make serial sections, which were stained for HE, Hu-mito, Hu-ki67, AR, PSA, CK8, CK5 and p63. Both low (i.e., 100x) and high-power (i.e., 400x) magnifications were shown. Arrow indicates CK5+ cells.
Figure 3.
IHC analysis of HPCa58 patient sample (GS7) and its xenograft tumor.
(A) Staining of HE, AR, PSA, CK8, CK5, p63 and Racamase in HPCa58 patient sample. (B) HPCa58P1 xenograft tumor, derived from coinjection with 100,000 Hs5 cells, was used to make serial sections were stained for HE, Hu-mito, Hu-ki67, AR, PSA, CK8, CK5 and p63. Both low (i.e., 100x) and high-power (i.e., 400x) magnifications were shown. Arrow indicates CK5+ cells.
Figure 4.
RT-PCR (A) and Western blotting (B–E) characterizations of HPCa/Hs5 xenograft tumors.
(A) RT-PCR results of AR, PSA, and β-actin using human-specific primers. LNCaP, TRPC (treatment-refractory prostate cancer; 46) and Du145 cells were used as controls. (B–D) Western blotting of Racemase, PSA, p63, CK18 and AR on HPCa-Hs5 xenograft tumors and the tumor cell-derived spheres. GAPDH and β-actin were used as loading controls. The arrow in D indicated the CK18 protein band (note that Hs5 cells and Hs5 tumor had some non-specific lower M.W bands). (E) Western blotting of AR, CK18 and β-actin in HPCa57/Hs5 serial xenograft tumors (P1–P4) from either subcutaneous or orthotopical (DP) injections. LNCaP and Du145 cells were used as controls.
Figure 5.
Cytogenetic analysis of HPCa/Hs5 xenograft tumors.
(A) An example of Hs5 cell karyotype. (B–C) Karyotypes of xenograft cells derived from HPCa70 piece implantation (B) or from HPCa70/Hs5 coinjections (C).
Figure 6.
Functional characterizations of EpCAM+ cells in HPCa/Hs5 tumors.
(A) FACS analysis of EpCAM expression in cultured Hs5 cells and tumor cells harvested from HPCa58P/Hs5 tumors (P5). (B) HE staining of tumors derived from 100 EpCAM+ and EpCAM− cells from HPCa58/Hs5 tumors. (C) Scheme of sphere formation assays using HPCa/Hs5 xenograft tumor cells. (D) Representative images of spheres derived from EpCAM+ HPCa84 and HPCa87 tumor cells (100x).
Figure 7.
Histological and cellular heterogeneity in GS7 prostate tumors.
Shown are whole-mount Aperio ScanScope images of HPCa57 (A) and HPCa58 (B) patient tumors, in which benign (B) glands, differentiated (D), and undifferentiated (U) tumor areas can be identified. Enlarged images of one differentiated and two undifferentiated areas are shown on the right. The magnifications of the original objectives are indicated.
Figure 8.
Histological and cellular heterogeneity in GS7 and GS9 prostate tumors.
Shown are whole-mount Aperio ScanScope images of HPCa70 (A) and HPCa101 (B) patient tumors, in which benign (B) glands, differentiated (D), and undifferentiated (U) tumor areas can be identified. Enlarged images of one differentiated and two undifferentiated areas are shown on the right. The magnifications of the original objectives are indicated.
Figure 9.
Histological analysis of HPCa70 patient sample (A) and its piece implant-derived xenograft tumor (B). Molecules stained and original magnifications are indicated.
Figure 10.
Histological analysis of HPCa101 patient sample (A) and its piece implant-derived xenograft tumor (B).
Molecules stained and original magnifications are indicated.