scholarly journals Expression of Piwi Genes during the Regeneration of Lineus sanguineus (Nemertea, Pilidiophora, Heteronemertea)

Genes ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1484
Author(s):  
Cong-Mei Xu ◽  
Shi-Chun Sun
Keyword(s):  
Stem Cells ◽  
Proliferating Cell Nuclear Antigen ◽  
Direct Evidence ◽  
Nuclear Antigen ◽  
Regeneration Capacity ◽  
Pluripotent Cells ◽  
Self Renewal ◽  
Proliferating Cell ◽  
Anterior Regeneration

The transposon silencer piwi genes play important roles in germline determination and maintenance, gametogenesis, and stem-cell self-renewal, and the expression of certain piwi genes is indispensable for regeneration. Knowledge about piwi genes is needed for phylum Nemertea, which contains members (e.g., Lineus sanguineus) with formidable regeneration capacity. By searching the L. sanguineus genome, we identified six Argonaute genes including three ago (Ls-Ago2, Ls-Ago2a, and Ls-Ago2b) and three piwi (Ls-piwi1, Ls-piwi2, and Ls-piwi3) genes. In situ hybridization revealed that, in intact females, Ls-piwi2 and Ls-piwi3 were not expressed, while Ls-piwi1 was expressed in ovaries. During regeneration, Ls-piwi1 and Ls-pcna (proliferating cell nuclear antigen) had strong and similar expressions. The expression of Ls-piwi1 became indetectable while Ls-pcna continued to be expressed when the differentiation of new organs was finished. During anterior regeneration, expression signals of Ls-piwi2 and Ls-piwi3 were weak and only detected in the blastema stage. During posterior regeneration, no expression was observed for Ls-piwi2. To date, no direct evidence has been found for the existence of congenital stem cells in adult L. sanguineus. The “pluripotent cells” in regenerating tissues are likely to be dedifferentiated from other type(s) of cells.

scholarly journals In Situ Analysis of Cellular Proliferation in Canine, Feline and Equine Tumors by Immunohistochemistry: A Comparison of Bromodeoxyuridine, Proliferating Cell Nuclear Antigen, and Interchromatin-Associated Antigen Immunostaining Techniques

1994 ◽  
Vol 6 (4) ◽  
pp. 453-457 ◽  
Author(s):  
Alain Pierre Théon ◽  
Loretta Metzger ◽  
Stephen Griffey
Keyword(s):  
Cell Proliferation ◽  
Proliferating Cell Nuclear Antigen ◽  
Cellular Proliferation ◽  
Nuclear Antigen ◽  
Brdu Incorporation ◽  
Immunohistochemical Detection ◽  
Proliferating Cells ◽  
Proliferating Cell

Cell proliferation in canine, feline, and equine tumors was evaluated using immunohistochemical detection of in vitro 5–bromodeoxyuridine (BrdU) incorporation, proliferating cell nuclear antigen (PCNA), and interchromatin-associated antigen (p105). Ten tumors in each species were analyzed. The tumor proliferative fraction (PF) was defined as the percentage of labeled nuclei for 5,000 tumor nuclei counted. Immunoreactivity was observed with all techniques in all species. A good correlation was observed between the proliferative fractions measured with the BrdU (PFBrdU) and PCNA (PFPCNA) techniques ( rs = 0.523, P = 0.0026). There was no correlation between the PFs measured with the BrdU (PFBrdU) and p105 (PFP105) techniques. Using the median values obtained from the different approaches as cutoff points to define slowly and rapidly proliferating tumors, there was an 80% agreement ( P = 0.009) between PFBrdU and PFPCNA and no agreement between PFBrdU and PFP105 The results of this study indicate that both BrdU and PCNA labeling methods can be used reliably for identifying proliferating cells in animal tumors. In addition, PCNA could be used to replace the BrdU method to assess tumor proliferative fraction because it does not require pretreatment of tissues.

Determination of cell proliferative activity by immnunohistochemical detection of proliferating cell nuclear antigen in N‐methyl nitrosoureainduced rat mammary tumours

2013 ◽  
Vol 3 (4) ◽  
pp. 188-194
Author(s):  
R. Barathidasan ◽  
R. V. S. Pawaiya ◽  
R. B. Rai ◽  
K. Dhama
Keyword(s):  
Papillary Carcinoma ◽  
Proliferating Cell Nuclear Antigen ◽  
Nuclear Antigen ◽  
Nuclear Staining ◽  
Chemical Carcinogen ◽  
Malignant Tumours ◽  
Mammary Tumours ◽  
Proliferating Cell

Rodent experimental models are considered useful experimental systems to study mammary cancer, which occurs in rodents as a result of hormonal imbalance, much like the breast cancer in women. Rodent cells are easier to transform by chemical carcinogen due to their relatively poorer controls of genetic stability and DNA repair systems. Proliferating cell nuclear antigen (PCNA) is one of the most sensitive biomarkers to measure the cell proliferative activities in the neoplasms, especially when the otherwise undetectable PCNA protein over‐expresses to the immunohistochemically detectable levels. In the present study, mammary tumours were induced in Sprague Dawley rats by intraperitoneal administration of N‐methyl nitrosourea (MNU) chemical carcinogen. Palpable tumours were noticed after 60 days postinjection (dpi) with average latency period of 144 days, and 12 (46.15%) of 26 rats developed mammary tumours. A total of 18 tumours were diagnosed in 12 rats, including 2 cases (10%) of hyperplasia, 1 (5%) benign and 17 malignant tumours (85%). Malignant tumours included in situ ductal papillary carcinoma (11.11%), in situ ductal cribriform carcinoma (38.88%), invasive tubular adenocarcinoma (5.56%), invasive papillary carcinoma (16.67%) and invasive cribriform carcinoma (22.22%) were. PCNA immunohistochemistry revealed moderate to strong nuclear staining in neoplastic cells, with highest mean PCNA index of 55.53±10.13 in invasive cribriform carcinoma. The PCNA indices were significantly different (p<0.01; one way ANOVA) between control mammary gland, hyperplasia, benign and malignant tumours. It was concluded that the measurement of PCNA expressing cells employing immunohistochemistry can help in the determination of the level of malignancy in MNU‐induced rat mammary tumours.

scholarly journals PAF promotes stemness and radioresistance of glioma stem cells

2017 ◽  
Vol 114 (43) ◽  
pp. E9086-E9095 ◽  
Author(s):  
Derrick Sek Tong Ong ◽  
Baoli Hu ◽  
Yan Wing Ho ◽  
Charles-Etienne Gabriel Sauvé ◽  
Christopher A. Bristow ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dna Replication ◽  
Therapeutic Option ◽  
Nuclear Antigen ◽  
Glioma Stem Cells ◽  
Cell Frequency ◽  
Self Renewal ◽  
Dna Damage Signaling ◽  
Proliferating Cell ◽  
Tumor Initiating Cell

An integrated genomic and functional analysis to elucidate DNA damage signaling factors promoting self-renewal of glioma stem cells (GSCs) identified proliferating cell nuclear antigen (PCNA)-associated factor (PAF) up-regulation in glioblastoma. PAF is preferentially overexpressed in GSCs. Its depletion impairs maintenance of self-renewal without promoting differentiation and reduces tumor-initiating cell frequency. Combined transcriptomic and metabolomic analyses revealed that PAF supports GSC maintenance, in part, by influencing DNA replication and pyrimidine metabolism pathways. PAF interacts with PCNA and regulates PCNA-associated DNA translesion synthesis (TLS); consequently, PAF depletion in combination with radiation generated fewer tumorspheres compared with radiation alone. Correspondingly, pharmacological impairment of DNA replication and TLS phenocopied the effect of PAF depletion in compromising GSC self-renewal and radioresistance, providing preclinical proof of principle that combined TLS inhibition and radiation therapy may be a viable therapeutic option in the treatment of glioblastoma multiforme (GBM).

scholarly journals HDAC2 and PCNA expression is correlated to decreasing of endoxifen sensitivity in human breast cancer stem cells ALDH+

2019 ◽  
Vol 10 (2) ◽  
pp. 77-81
Author(s):  
Syarifah Dewi ◽  
Mohamad Sadikin ◽  
Muchlis Ramli ◽  
Septelia Inawati Wanandi
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Proliferating Cell Nuclear Antigen ◽  
Health Science ◽  
Nuclear Antigen ◽  
Breast Cancer Stem Cells ◽  
Human Breast ◽  
Histone Deacetylase 2 ◽  
Proliferating Cell

Latar belakang: Sel punca kanker payudara (breast cancer stem cells/BCSC) adalah subpopulasi sel kanker yang memiliki kemampuan menghasilkan tumor baru dan bersifat seperti sel punca. Penelitian kami sebelumnya menggunakan jaringan kanker payudara mengungkapkan bahwa ekspresi gen histone deacetylase 2 (HDAC2) dan proliferating cell nuclear antigen (PCNA) ditemukan perbedaan signifikan setelah terapi neoajuvan hormon dan kemoterapi. Penelitian ini bertujuan untuk menganalisis hubungan antara ekspresi HDAC2 dan PCNA dengan kelangsungan hidup sel punca kanker payudara dengan penanda aldehyde dehydrogenase + (ALDH+) yang diberi perlakuan endoksifen. Metode: Sampel adalah BCSC primer manusia ALDH+ yang diberi perlakuan endoksifen 4 uM masingmasing selama 2, 4, 6, 8, 10, 12, 14 hari. Viabilitas sel dilihat dengan menggunakan trypan blue dan ekspresi mRNA HDAC2 dan PCNA ditentukan menggunakan qRT-PCR. Hasil: Viabilitas BCSCs ALDH + menurun setelah 2 sampai 4 hari pemberian endoksifen. Pada periode ini juga didapatkan ekspresi mRNA HDAC2 dan PCNA mengalami penurunan. Tetapi setelah pemberian endoksifen selama 8 hari, viabilitas BCSCs ALDH + mengalami peningkatan dan ditemukan peningkatan yang signifikan pada hari ke-14 pemberian endoksifen. Ekspresi mRNA HDAC2 dan PCNA juga menunjukkan peningkatan mulai pada hari ke-8 dan terus meningkat hingga hari ke-14 pemberian endoksifen. Penelitian ini menunjukkan pola yang sama antara ekspresi mRNA HDAC2 dan PCNA dan viabilitas sel. Kesimpulan: Induksi endoksifen yang lama menurunkan sensitivitas efek endoksifen pada BCSC manusia dan ekspresi HDAC2 dan PCNA berkorelasi dengan viabilitas BCSC manusia setelah induksi endoksifen. (Health Science Journal of Indonesia 2019;10(2):77-81) Kata kunci: sel punca kanker payudara, viabilitas sel, HDAC2, PCNA, endoksifen   Abstract Background: Breast cancer stem cells (BCSCs) are subpopulation of cancer cells that has the ability to generate new tumor and similar properties to stem cell. Our previous study using breast cancer patients revealed that gene expression of histone deacetylase 2 (HDAC2) and proliferating cell nuclear antigen (PCNA) were significantly altered after neoadjuvant hormone and chemotherapy. This study aimed to analyze the correlation between HDAC2 and PCNA expressions with the viability of breast cancer stem cells aldehyde dehydrogenase + (BCSC ALDH+) treated by endoxifen. Method: Samples are human primary BCSCs ALDH+ that treated with 4 uM of endoxifen for 2, 4, 6, 8, 10, 12, 14 days, respectively. Cell viability was measured using trypan blue exclusion assay and the mRNA expressions of HDAC2 and PCNA were determined using qRT-PCR. Results: The viability of BCSCs ALDH+ was decreased after 2 days until 4 days-endoxifen treatment. It also demonstrated that mRNA expression of HDAC2 and PCNA were decreased in this period. But after 8 days endoxifen treatment, the viability of BCSCs ALDH+ was increased. The increasing of viability was higher in 14 days-endoxifen treatment. The mRNA expression of HDAC2 and PCNA also showed increasing begin on 8 days and continued to increase until 14-days endoxifen treatment. We found a similar pattern between HDAC2 and PCNA expression and cell viability. Conclusion: Prolonge endoxifen treatment decrease sensitivity of endoxifen effect in human BCSC and the expression of HDAC2 and PCNA are correlated to human BCSCs viability after endoxifen treatment. (Health Science Journal of Indonesia 2019;10(2):77-81) Keywords: human breast cancer stem cells, viability, HDAC2, PCNA, endoxifen

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