Journal: The World Journal of Men's Health

Article Title: Human Adipose Stem Cells Engineered to Express Carboxylesterase Confer Anti-Tumoral Efficacy of Irinotecan in Castration-Resistant Prostate Cancer Bone Metastasis Growth and Osteolysis

doi: 10.5534/wjmh.240284

Figure Lengend Snippet: Migration of CE-expressing ADSC to prostate cancer cell cultures, potentially mediated by chemoattraction. (A, B) Cell migration assay using Matrigel™-coated Transwell chambers. hTERT-ADSC, hTERT-ADSC-CE1, and hTERT-ADSC-CE2 cells were plated in the upper chambers and co-cultured with empty media, WPMY-1 (WPM, human prostate myofibroblast cell line) or PC3 (human prostate cancer cell line) in the bottom chambers, as indicated. Migrated cells were stained, photographed (A) and quantified (B). (C–E) Expression of chemoattractant factors such as vascular endothelial growth factor (VEGF), stem cell factor (SCF) and stromal cell-derived factor 1 (SDC-1) and the corresponding receptors c-KIT, chemokine receptor 4 (CXCR4), VEGF receptors (VEGFR)-1, -2 and -3. Values are presented as mean±standard error of the mean. (A, B) ** p<0.01 compared with the ADSC/Media group. ## p<0.01 compared with the ADSC/WPMY-1 group. $$ p<0.01 compared with the ADSC/PC3 group. AA p<0.01 compared with the ADSC.CE1/PC3 group. BB p<0.01 compared with the ADSC.CE2/PC3 group. (C–E) * p<0.05 and ** p<0.01 compared with PC3. # p<0.05 and ## p<0.01 compared with ADSC, $$ p<0.01 compared with ADSC.CE1.CE: carboxylesterase, ADSC: adipose-derived stem cells, hTERT: human telomere reverse transcriptase gene, CPT-11: irinotecan.

Article Snippet: Similarly, human prostate adenocarcinoma cells (PC3), were bought from the Korean Cell Line Bank and cultured using the same protocol described above.

Techniques: Migration, Expressing, Cell Migration Assay, Cell Culture, Staining, Derivative Assay, Reverse Transcription

Journal: The World Journal of Men's Health

Article Title: Human Adipose Stem Cells Engineered to Express Carboxylesterase Confer Anti-Tumoral Efficacy of Irinotecan in Castration-Resistant Prostate Cancer Bone Metastasis Growth and Osteolysis

doi: 10.5534/wjmh.240284

Figure Lengend Snippet: Co-culture of CE-expressing ADSC confers cytotoxic effects of CPT-11 on PC3 cells. (A) The cytotoxic effects of CPT-11 (0.1 µM) on hTERT-ADSC. GFP, hTERT-ADSC.CE1, and hTERT-ADSC.CE2 were measured using the MTT cell viability assay. CE expression significantly increased the sensitivity to CPT-11. (B, C) Co-culture of hTERT-ADSC.CE1 or hTERT-ADSC.CE2 enhanced the cytotoxic effects of CPT-11 on PC3 cells, as measured by the MTT cell viability assay (B) and flow cytometry-based apoptosis assay (C). Values are presented as mean±standard error of the mean. ** p<0.01 compared with the PC3 alone group. ## p<0.01 compared with the PC3 plus CPT-11 5 µM group. $$ p<0.01 compared with the ADSC/PC3 group. A p<0.05 compared with the PC3 alone group. AA p<0.01 compared with the ADSC.CE1/PC3 group. CE: carboxylesterase, ADSC: adipose-derived stem cells, hTERT: human telomere reverse transcriptase gene, CPT-11: irinotecan.

Article Snippet: Similarly, human prostate adenocarcinoma cells (PC3), were bought from the Korean Cell Line Bank and cultured using the same protocol described above.

Techniques: Co-Culture Assay, Expressing, Viability Assay, Flow Cytometry, Apoptosis Assay, Derivative Assay, Reverse Transcription

Journal: The World Journal of Men's Health

Article Title: Human Adipose Stem Cells Engineered to Express Carboxylesterase Confer Anti-Tumoral Efficacy of Irinotecan in Castration-Resistant Prostate Cancer Bone Metastasis Growth and Osteolysis

doi: 10.5534/wjmh.240284

Figure Lengend Snippet: Co-implantation of CE-expressing ADSC with PC3 prostate cancer cells increase the anti-cancer effects of CPT-11 on prostate cancer bone metastasis. To evaluate the therapeutic efficacy of hTERT-ADSC.CE2 combined with CPT-11 in a bone metastasis mouse model, in vivo experiments were conducted to assess tumor growth inhibition and osteolysis reduction. Male athymic mice (4–5 weeks old, n=10/group) were injected with PC3 prostate cancer cells (2×10 4 cells) into the proximal tibia, followed by hTERT-ADSC.CE2 cell injection (1×10 5 cells) into the left heart ventricle to model systemic dissemination. CPT-11 (1.7 mg/kg) was administered intraperitoneally starting on day 3, following a regimen of five consecutive days of treatment with two-day breaks, for a total of three weeks. (A, B) Bioluminescence imaging (A) and X-ray images (B) of representative mice in each group showing the tumor growth and osteolysis in bone. (C) Histologic images of the tumors (×20). CE: carboxylesterase, ADSC: adipose-derived stem cells, hTERT: human telomere reverse transcriptase gene, CPT-11: irinotecan, H&E: hematoxylin and eosin staining, TRAP: tartrate-resistant acid phosphatase, CK8: cytokeratin 8.

Article Snippet: Similarly, human prostate adenocarcinoma cells (PC3), were bought from the Korean Cell Line Bank and cultured using the same protocol described above.

Techniques: Expressing, Drug discovery, In Vivo, Inhibition, Injection, Imaging, Derivative Assay, Reverse Transcription, Staining