Co-staining of microRNAs and their target proteins by miRNA in situ hybridization and immunohistofluorescence on prostate cancer tissue microarrays
The co-expression of miRNAs and their target proteins was studied on tissue microarrays from different prostate cancer (PCa) patients. PCa of primary Gleason pattern 4 (GP4), lymph node metastases of GP4, distant metastases, and normal tissue from the transitional and peripheral zones were co-staine...
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| Veröffentlicht in: | Laboratory investigation 2019-10, Vol.99 (10), p.1527-1534 |
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| creator | Eckstein, Markus Sailer, Verena Nielsen, Boye Schnack Wittenberg, Thomas Wiesmann, Veit Lieb, Verena Nolte, Elke Hartmann, Arndt Kristiansen, Glen Wernert, Nicolas Wullich, Bernd Taubert, Helge Wach, Sven |
| description | The co-expression of miRNAs and their target proteins was studied on tissue microarrays from different prostate cancer (PCa) patients. PCa of primary Gleason pattern 4 (GP4), lymph node metastases of GP4, distant metastases, and normal tissue from the transitional and peripheral zones were co-stained by fluorescent miRNA in situ hybridization (miRisH) and protein immunohistofluorescence (IHF). The miRNAs and corresponding target proteins include the pairs miR-145/ERG, miR-143/uPAR, and miR-375/SEC23A. The fluorescence-stained and scanned tissue microarrays (TMAs) were evaluated by experienced uropathologists. The pair miR-145/ERG showed an exclusive staining for miR-145 in the nuclei of stromal cells, both in tumor and normal tissue, and for ERG in the cytoplasm with/without co-expression in the nucleus of tumor cells. The pair miR-143/uPAR revealed a clear distinction between miR-143 in the nuclei of stromal cells and uPAR staining in the cytoplasm of tumor cells. Metastases (lymph node and distant) however, showed tumor cells with cytoplasmic staining for miR-143/uPAR. In normal tissues, beside the nuclei of the stroma cells, gland cells could also express miR-143 and uPAR in the cytoplasm. miR-375 showed particular staining in the nucleoli of GP4 and metastatic samples, suggesting that nucleoli play a special role in sequestering proteins and miRNAs. Combined miRisH/IHF allows for the study of miRNA expression patterns and their target proteins at the single-cell level. |
| doi_str_mv | 10.1038/s41374-019-0251-8 |
| format | Article |
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PCa of primary Gleason pattern 4 (GP4), lymph node metastases of GP4, distant metastases, and normal tissue from the transitional and peripheral zones were co-stained by fluorescent miRNA in situ hybridization (miRisH) and protein immunohistofluorescence (IHF). The miRNAs and corresponding target proteins include the pairs miR-145/ERG, miR-143/uPAR, and miR-375/SEC23A. The fluorescence-stained and scanned tissue microarrays (TMAs) were evaluated by experienced uropathologists. The pair miR-145/ERG showed an exclusive staining for miR-145 in the nuclei of stromal cells, both in tumor and normal tissue, and for ERG in the cytoplasm with/without co-expression in the nucleus of tumor cells. The pair miR-143/uPAR revealed a clear distinction between miR-143 in the nuclei of stromal cells and uPAR staining in the cytoplasm of tumor cells. Metastases (lymph node and distant) however, showed tumor cells with cytoplasmic staining for miR-143/uPAR. In normal tissues, beside the nuclei of the stroma cells, gland cells could also express miR-143 and uPAR in the cytoplasm. miR-375 showed particular staining in the nucleoli of GP4 and metastatic samples, suggesting that nucleoli play a special role in sequestering proteins and miRNAs. Combined miRisH/IHF allows for the study of miRNA expression patterns and their target proteins at the single-cell level.</description><identifier>ISSN: 0023-6837</identifier><identifier>EISSN: 1530-0307</identifier><identifier>DOI: 10.1038/s41374-019-0251-8</identifier><identifier>PMID: 31186527</identifier><language>eng</language><publisher>New York: Elsevier Inc</publisher><subject>13/51 ; 14/32 ; 631/67/1857 ; 692/53/2422 ; 82/80 ; 96/63 ; Cytoplasm ; Fluorescence ; Fluorescent Antibody Technique - methods ; Humans ; Hybridization ; In Situ Hybridization, Fluorescence - methods ; Laboratory Medicine ; Lymph nodes ; Lymphatic system ; Male ; Medicine ; Medicine & Public Health ; Metastases ; Metastasis ; MicroRNAs ; MicroRNAs - analysis ; MicroRNAs - metabolism ; miRNA ; Nuclei ; Nuclei (cytology) ; Nucleoli ; Pathology ; Prostate cancer ; Prostatic Neoplasms - chemistry ; Prostatic Neoplasms - metabolism ; Proteins ; Sequestering ; Staining ; Stromal cells ; technical-report ; Tissue Array Analysis ; Tissues ; Tumor cells ; Tumors</subject><ispartof>Laboratory investigation, 2019-10, Vol.99 (10), p.1527-1534</ispartof><rights>2019 United States & Canadian Academy of Pathology</rights><rights>United States & Canadian Academy of Pathology 2019</rights><rights>United States & Canadian Academy of Pathology 2019.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c467t-952d248f80d9ee1d801b564514c5360366e436b66e95411e2c2a00bc5067c8773</citedby><cites>FETCH-LOGICAL-c467t-952d248f80d9ee1d801b564514c5360366e436b66e95411e2c2a00bc5067c8773</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31186527$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Eckstein, Markus</creatorcontrib><creatorcontrib>Sailer, Verena</creatorcontrib><creatorcontrib>Nielsen, Boye Schnack</creatorcontrib><creatorcontrib>Wittenberg, Thomas</creatorcontrib><creatorcontrib>Wiesmann, Veit</creatorcontrib><creatorcontrib>Lieb, Verena</creatorcontrib><creatorcontrib>Nolte, Elke</creatorcontrib><creatorcontrib>Hartmann, Arndt</creatorcontrib><creatorcontrib>Kristiansen, Glen</creatorcontrib><creatorcontrib>Wernert, Nicolas</creatorcontrib><creatorcontrib>Wullich, Bernd</creatorcontrib><creatorcontrib>Taubert, Helge</creatorcontrib><creatorcontrib>Wach, Sven</creatorcontrib><creatorcontrib>On behalf of the German Prostate Cancer Consortium (DPKK)</creatorcontrib><creatorcontrib>German Prostate Cancer Consortium (DPKK)</creatorcontrib><title>Co-staining of microRNAs and their target proteins by miRNA in situ hybridization and immunohistofluorescence on prostate cancer tissue microarrays</title><title>Laboratory investigation</title><addtitle>Lab Invest</addtitle><addtitle>Lab Invest</addtitle><description>The co-expression of miRNAs and their target proteins was studied on tissue microarrays from different prostate cancer (PCa) patients. PCa of primary Gleason pattern 4 (GP4), lymph node metastases of GP4, distant metastases, and normal tissue from the transitional and peripheral zones were co-stained by fluorescent miRNA in situ hybridization (miRisH) and protein immunohistofluorescence (IHF). The miRNAs and corresponding target proteins include the pairs miR-145/ERG, miR-143/uPAR, and miR-375/SEC23A. The fluorescence-stained and scanned tissue microarrays (TMAs) were evaluated by experienced uropathologists. The pair miR-145/ERG showed an exclusive staining for miR-145 in the nuclei of stromal cells, both in tumor and normal tissue, and for ERG in the cytoplasm with/without co-expression in the nucleus of tumor cells. The pair miR-143/uPAR revealed a clear distinction between miR-143 in the nuclei of stromal cells and uPAR staining in the cytoplasm of tumor cells. Metastases (lymph node and distant) however, showed tumor cells with cytoplasmic staining for miR-143/uPAR. In normal tissues, beside the nuclei of the stroma cells, gland cells could also express miR-143 and uPAR in the cytoplasm. miR-375 showed particular staining in the nucleoli of GP4 and metastatic samples, suggesting that nucleoli play a special role in sequestering proteins and miRNAs. Combined miRisH/IHF allows for the study of miRNA expression patterns and their target proteins at the single-cell level.</description><subject>13/51</subject><subject>14/32</subject><subject>631/67/1857</subject><subject>692/53/2422</subject><subject>82/80</subject><subject>96/63</subject><subject>Cytoplasm</subject><subject>Fluorescence</subject><subject>Fluorescent Antibody Technique - methods</subject><subject>Humans</subject><subject>Hybridization</subject><subject>In Situ Hybridization, Fluorescence - methods</subject><subject>Laboratory Medicine</subject><subject>Lymph nodes</subject><subject>Lymphatic system</subject><subject>Male</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Metastases</subject><subject>Metastasis</subject><subject>MicroRNAs</subject><subject>MicroRNAs - analysis</subject><subject>MicroRNAs - metabolism</subject><subject>miRNA</subject><subject>Nuclei</subject><subject>Nuclei (cytology)</subject><subject>Nucleoli</subject><subject>Pathology</subject><subject>Prostate cancer</subject><subject>Prostatic Neoplasms - chemistry</subject><subject>Prostatic Neoplasms - metabolism</subject><subject>Proteins</subject><subject>Sequestering</subject><subject>Staining</subject><subject>Stromal cells</subject><subject>technical-report</subject><subject>Tissue Array Analysis</subject><subject>Tissues</subject><subject>Tumor cells</subject><subject>Tumors</subject><issn>0023-6837</issn><issn>1530-0307</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kU2LFDEQhoMo7rj6A7xIwIuX1nx3D56WwS9YFETPIZ2unskynaxJemH8G_5hq7dXBQ97Kkg971uVegl5ztlrzmT3piguW9Uwvm2Y0LzpHpAN15I1TLL2IdkwJmRjOtmekSelXDHGlTL6MTmTnHdGi3ZDfu1SU6oLMcQ9TSOdgs_p6-eLQl0caD1AyLS6vIdKr3OqEGKh_QkxZGiItIQ608Opz2EIP10NKd4KwzTNMR1CqWk8zilD8RA9UGyjDQ6sQL3DF3QPpcywDnY5u1N5Sh6N7ljg2V09J9_fv_u2-9hcfvnwaXdx2Xhl2tpstRiE6saODVsAPnSM99oozZXX0jBpDChpeixbrTgH4YVjrPeamdZ3bSvPyavVF1f6MUOpdgq45_HoIqS5WCG2xixnkoi-_A-9SnOOuN0txZlSejHkK4VfKSXDaK9zmFw-Wc7skphdE7OYmF0Ssx1qXtw5z_0Ew1_Fn4gQECtQsBX3kP-Nvs_17SoCvN9NQFHxYUlgCBl8tUMK96h_A4j3tXI</recordid><startdate>20191001</startdate><enddate>20191001</enddate><creator>Eckstein, Markus</creator><creator>Sailer, Verena</creator><creator>Nielsen, Boye Schnack</creator><creator>Wittenberg, Thomas</creator><creator>Wiesmann, Veit</creator><creator>Lieb, Verena</creator><creator>Nolte, Elke</creator><creator>Hartmann, Arndt</creator><creator>Kristiansen, Glen</creator><creator>Wernert, Nicolas</creator><creator>Wullich, Bernd</creator><creator>Taubert, Helge</creator><creator>Wach, Sven</creator><general>Elsevier Inc</general><general>Nature Publishing Group US</general><general>Nature Publishing Group</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7T5</scope><scope>7T7</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope></search><sort><creationdate>20191001</creationdate><title>Co-staining of microRNAs and their target proteins by miRNA in situ hybridization and immunohistofluorescence on prostate cancer tissue microarrays</title><author>Eckstein, Markus ; Sailer, Verena ; Nielsen, Boye Schnack ; Wittenberg, Thomas ; Wiesmann, Veit ; Lieb, Verena ; Nolte, Elke ; Hartmann, Arndt ; Kristiansen, Glen ; Wernert, Nicolas ; Wullich, Bernd ; Taubert, Helge ; Wach, Sven</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c467t-952d248f80d9ee1d801b564514c5360366e436b66e95411e2c2a00bc5067c8773</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>13/51</topic><topic>14/32</topic><topic>631/67/1857</topic><topic>692/53/2422</topic><topic>82/80</topic><topic>96/63</topic><topic>Cytoplasm</topic><topic>Fluorescence</topic><topic>Fluorescent Antibody Technique - methods</topic><topic>Humans</topic><topic>Hybridization</topic><topic>In Situ Hybridization, Fluorescence - methods</topic><topic>Laboratory Medicine</topic><topic>Lymph nodes</topic><topic>Lymphatic system</topic><topic>Male</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Metastases</topic><topic>Metastasis</topic><topic>MicroRNAs</topic><topic>MicroRNAs - analysis</topic><topic>MicroRNAs - metabolism</topic><topic>miRNA</topic><topic>Nuclei</topic><topic>Nuclei (cytology)</topic><topic>Nucleoli</topic><topic>Pathology</topic><topic>Prostate cancer</topic><topic>Prostatic Neoplasms - chemistry</topic><topic>Prostatic Neoplasms - metabolism</topic><topic>Proteins</topic><topic>Sequestering</topic><topic>Staining</topic><topic>Stromal cells</topic><topic>technical-report</topic><topic>Tissue Array Analysis</topic><topic>Tissues</topic><topic>Tumor cells</topic><topic>Tumors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Eckstein, Markus</creatorcontrib><creatorcontrib>Sailer, Verena</creatorcontrib><creatorcontrib>Nielsen, Boye Schnack</creatorcontrib><creatorcontrib>Wittenberg, Thomas</creatorcontrib><creatorcontrib>Wiesmann, Veit</creatorcontrib><creatorcontrib>Lieb, Verena</creatorcontrib><creatorcontrib>Nolte, Elke</creatorcontrib><creatorcontrib>Hartmann, Arndt</creatorcontrib><creatorcontrib>Kristiansen, Glen</creatorcontrib><creatorcontrib>Wernert, Nicolas</creatorcontrib><creatorcontrib>Wullich, Bernd</creatorcontrib><creatorcontrib>Taubert, Helge</creatorcontrib><creatorcontrib>Wach, Sven</creatorcontrib><creatorcontrib>On behalf of the German Prostate Cancer Consortium (DPKK)</creatorcontrib><creatorcontrib>German Prostate Cancer Consortium (DPKK)</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>MEDLINE - Academic</collection><jtitle>Laboratory investigation</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Eckstein, Markus</au><au>Sailer, Verena</au><au>Nielsen, Boye Schnack</au><au>Wittenberg, Thomas</au><au>Wiesmann, Veit</au><au>Lieb, Verena</au><au>Nolte, Elke</au><au>Hartmann, Arndt</au><au>Kristiansen, Glen</au><au>Wernert, Nicolas</au><au>Wullich, Bernd</au><au>Taubert, Helge</au><au>Wach, Sven</au><aucorp>On behalf of the German Prostate Cancer Consortium (DPKK)</aucorp><aucorp>German Prostate Cancer Consortium (DPKK)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Co-staining of microRNAs and their target proteins by miRNA in situ hybridization and immunohistofluorescence on prostate cancer tissue microarrays</atitle><jtitle>Laboratory investigation</jtitle><stitle>Lab Invest</stitle><addtitle>Lab Invest</addtitle><date>2019-10-01</date><risdate>2019</risdate><volume>99</volume><issue>10</issue><spage>1527</spage><epage>1534</epage><pages>1527-1534</pages><issn>0023-6837</issn><eissn>1530-0307</eissn><abstract>The co-expression of miRNAs and their target proteins was studied on tissue microarrays from different prostate cancer (PCa) patients. PCa of primary Gleason pattern 4 (GP4), lymph node metastases of GP4, distant metastases, and normal tissue from the transitional and peripheral zones were co-stained by fluorescent miRNA in situ hybridization (miRisH) and protein immunohistofluorescence (IHF). The miRNAs and corresponding target proteins include the pairs miR-145/ERG, miR-143/uPAR, and miR-375/SEC23A. The fluorescence-stained and scanned tissue microarrays (TMAs) were evaluated by experienced uropathologists. The pair miR-145/ERG showed an exclusive staining for miR-145 in the nuclei of stromal cells, both in tumor and normal tissue, and for ERG in the cytoplasm with/without co-expression in the nucleus of tumor cells. The pair miR-143/uPAR revealed a clear distinction between miR-143 in the nuclei of stromal cells and uPAR staining in the cytoplasm of tumor cells. Metastases (lymph node and distant) however, showed tumor cells with cytoplasmic staining for miR-143/uPAR. In normal tissues, beside the nuclei of the stroma cells, gland cells could also express miR-143 and uPAR in the cytoplasm. miR-375 showed particular staining in the nucleoli of GP4 and metastatic samples, suggesting that nucleoli play a special role in sequestering proteins and miRNAs. Combined miRisH/IHF allows for the study of miRNA expression patterns and their target proteins at the single-cell level.</abstract><cop>New York</cop><pub>Elsevier Inc</pub><pmid>31186527</pmid><doi>10.1038/s41374-019-0251-8</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | 13/51 14/32 631/67/1857 692/53/2422 82/80 96/63 Cytoplasm Fluorescence Fluorescent Antibody Technique - methods Humans Hybridization In Situ Hybridization, Fluorescence - methods Laboratory Medicine Lymph nodes Lymphatic system Male Medicine Medicine & Public Health Metastases Metastasis MicroRNAs MicroRNAs - analysis MicroRNAs - metabolism miRNA Nuclei Nuclei (cytology) Nucleoli Pathology Prostate cancer Prostatic Neoplasms - chemistry Prostatic Neoplasms - metabolism Proteins Sequestering Staining Stromal cells technical-report Tissue Array Analysis Tissues Tumor cells Tumors |
| title | Co-staining of microRNAs and their target proteins by miRNA in situ hybridization and immunohistofluorescence on prostate cancer tissue microarrays |
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