Expansion of mouse castration-resistant intermediate prostate stem cells in vitro
Abstract Background Most castration-resistant prostate cancers (CRPCs) have a luminal phenotype with high androgen receptor (AR) and prostate-specific antigen (PSA) expression. Currently, it is difficult to culture castration-resistant luminal cells with AR and PSA expression. Methods We formulated...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-07-01
|
| Series: | Stem Cell Research & Therapy |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13287-022-02978-x |
| _version_ | 1828541767381680128 |
|---|---|
| author | Yalan Xu Jie Mu Zhixia Zhou Yu Leng Yali Yu Xiuyue Song Aihua Liu Hai Zhu Jing Li Dong Wang |
| author_facet | Yalan Xu Jie Mu Zhixia Zhou Yu Leng Yali Yu Xiuyue Song Aihua Liu Hai Zhu Jing Li Dong Wang |
| author_sort | Yalan Xu |
| collection | DOAJ |
| description | Abstract Background Most castration-resistant prostate cancers (CRPCs) have a luminal phenotype with high androgen receptor (AR) and prostate-specific antigen (PSA) expression. Currently, it is difficult to culture castration-resistant luminal cells with AR and PSA expression. Methods We formulated a custom-made medium and isolated primary cells from the prostate of adult wild-type (WT) and TRAMP mice. The cells were characterized by immunofluorescence staining, transcriptomic analysis, and qRT-PCR verification. Their self-renewal and differentiation potential in vitro and in vivo were examined. We treated the cells with androgen deprivation and enzalutamide and performed immunofluorescence staining and western blotting to analyze their expression of AR and PSA. Results We isolated a novel type of castration-resistant intermediate prostate stem cells (CRIPSCs) from adult WT and TRAMP mice. The mouse CRIPSCs proliferated rapidly in two-dimensional (2D) culture dishes and can be cultured for more than six months. The mouse CRIPSCs expressed luminal markers (AR, PSA, and Dsg4), basal markers (CK5 and p63), Psca, and the intermediate cell marker (Ivl). Transcriptomic analysis showed that the mouse CRIPSCs had upregulated signaling pathways related to cancer development and drug resistance. In the long-term culture, TRAMP CRIPSCs had higher expression of the genes related to stem cells and cancers than WT mice. Both WT and TRAMP CRIPSCs formed organoids in Matrigel. WT CRIPSCs did not form prostate tissues when transplanted in vivo without urogenital sinus mesenchyme (UGM) cells. In contrast, TRAMP CRIPSCs formed prostate ducts in NOG mice without UGM cells and differentiated into luminal, basal, and neuroendocrine cells. Androgens regulated AR translocation between the nucleus and cytoplasm in the mouse CRIPSCs. Treatment of androgen deprivation (ADT) and enzalutamide reduced AR expression in WT and TRAMP CRIPSCs; however, this treatment promoted PSA expression in TRAMP, while not WT CRIPSCs, similar to the clinical observations of CRPC. Conclusions Our study established a method for isolating and expanding mouse CRIPSCs in 2D culture dishes. Mouse CRIPSCs had markers of basal and luminal cells, including AR and PSA, and can differentiate into prostate organoids and tissues. TRAMP CRIPSCs had elevated PSA expression upon ADT and enzalutamide treatment. Our method can be translated into clinical settings for CRPC precision medicine. |
| first_indexed | 2024-12-12T01:44:38Z |
| format | Article |
| id | doaj.art-9aad6d02b2b04e7b8b6d6c36589d1841 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-12T01:44:38Z |
| publishDate | 2022-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-9aad6d02b2b04e7b8b6d6c36589d18412022-12-22T00:42:37ZengBMCStem Cell Research & Therapy1757-65122022-07-0113111310.1186/s13287-022-02978-xExpansion of mouse castration-resistant intermediate prostate stem cells in vitroYalan Xu0Jie Mu1Zhixia Zhou2Yu Leng3Yali Yu4Xiuyue Song5Aihua Liu6Hai Zhu7Jing Li8Dong Wang9Institute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityCollege of Life Sciences, and School of Pharmacy, Medical College, Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityCollege of Life Sciences, and School of Pharmacy, Medical College, Qingdao UniversityDepartment of Urology, Qingdao Municipal Hospital Affiliated to Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityInstitute for Translational Medicine, The Affiliated Hospital of Qingdao University, Medical College, Qingdao UniversityAbstract Background Most castration-resistant prostate cancers (CRPCs) have a luminal phenotype with high androgen receptor (AR) and prostate-specific antigen (PSA) expression. Currently, it is difficult to culture castration-resistant luminal cells with AR and PSA expression. Methods We formulated a custom-made medium and isolated primary cells from the prostate of adult wild-type (WT) and TRAMP mice. The cells were characterized by immunofluorescence staining, transcriptomic analysis, and qRT-PCR verification. Their self-renewal and differentiation potential in vitro and in vivo were examined. We treated the cells with androgen deprivation and enzalutamide and performed immunofluorescence staining and western blotting to analyze their expression of AR and PSA. Results We isolated a novel type of castration-resistant intermediate prostate stem cells (CRIPSCs) from adult WT and TRAMP mice. The mouse CRIPSCs proliferated rapidly in two-dimensional (2D) culture dishes and can be cultured for more than six months. The mouse CRIPSCs expressed luminal markers (AR, PSA, and Dsg4), basal markers (CK5 and p63), Psca, and the intermediate cell marker (Ivl). Transcriptomic analysis showed that the mouse CRIPSCs had upregulated signaling pathways related to cancer development and drug resistance. In the long-term culture, TRAMP CRIPSCs had higher expression of the genes related to stem cells and cancers than WT mice. Both WT and TRAMP CRIPSCs formed organoids in Matrigel. WT CRIPSCs did not form prostate tissues when transplanted in vivo without urogenital sinus mesenchyme (UGM) cells. In contrast, TRAMP CRIPSCs formed prostate ducts in NOG mice without UGM cells and differentiated into luminal, basal, and neuroendocrine cells. Androgens regulated AR translocation between the nucleus and cytoplasm in the mouse CRIPSCs. Treatment of androgen deprivation (ADT) and enzalutamide reduced AR expression in WT and TRAMP CRIPSCs; however, this treatment promoted PSA expression in TRAMP, while not WT CRIPSCs, similar to the clinical observations of CRPC. Conclusions Our study established a method for isolating and expanding mouse CRIPSCs in 2D culture dishes. Mouse CRIPSCs had markers of basal and luminal cells, including AR and PSA, and can differentiate into prostate organoids and tissues. TRAMP CRIPSCs had elevated PSA expression upon ADT and enzalutamide treatment. Our method can be translated into clinical settings for CRPC precision medicine.https://doi.org/10.1186/s13287-022-02978-xcastration-resistant prostate cancerStem cellIntermediate cellAndrogen receptorProstate-specific antigenAndrogen deprivation |
| spellingShingle | Yalan Xu Jie Mu Zhixia Zhou Yu Leng Yali Yu Xiuyue Song Aihua Liu Hai Zhu Jing Li Dong Wang Expansion of mouse castration-resistant intermediate prostate stem cells in vitro Stem Cell Research & Therapy castration-resistant prostate cancer Stem cell Intermediate cell Androgen receptor Prostate-specific antigen Androgen deprivation |
| title | Expansion of mouse castration-resistant intermediate prostate stem cells in vitro |
| title_full | Expansion of mouse castration-resistant intermediate prostate stem cells in vitro |
| title_fullStr | Expansion of mouse castration-resistant intermediate prostate stem cells in vitro |
| title_full_unstemmed | Expansion of mouse castration-resistant intermediate prostate stem cells in vitro |
| title_short | Expansion of mouse castration-resistant intermediate prostate stem cells in vitro |
| title_sort | expansion of mouse castration resistant intermediate prostate stem cells in vitro |
| topic | castration-resistant prostate cancer Stem cell Intermediate cell Androgen receptor Prostate-specific antigen Androgen deprivation |
| url | https://doi.org/10.1186/s13287-022-02978-x |
| work_keys_str_mv | AT yalanxu expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT jiemu expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT zhixiazhou expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT yuleng expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT yaliyu expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT xiuyuesong expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT aihualiu expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT haizhu expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT jingli expansionofmousecastrationresistantintermediateprostatestemcellsinvitro AT dongwang expansionofmousecastrationresistantintermediateprostatestemcellsinvitro |