• Clinical and Experimental Reproductive Medicine
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• v.36 no.3
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• pp.163-174
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• 2009

Objectives: Members of the immortalization-upregulated protein (IMUP) family are nuclear proteins implicated in SV40-mediated immortalization and cellular proliferation, but the mechanisms by which their expression is regulated are still unknown in placenta. To investigate to expression and functions of IMUPs in placenta, we conducted to compare IMUPs expression in normal and preeclamptic placenta tissues and analyzed the function of IMUP-2 in HTR-8/SVneo trophoblast cells after IMUP-2 gene transfection. Methods: The expression of IMUPs was analyzed in placental tissues from the following groups of patients (none underwent labor): 1) term normal placenta (n=15); 2) term with preeclamptic placeneta (n=15); and 3) pre-term with preeclamptic placenta (n=11) using semi-quantitative RT-PCR, RNA in situ hybiridization, immunohistochemistry, and Western blot. In order to evaluate the function of IMUP-2 in HTR-8/SVneo trophoblast cells, IMUP-2 plasmids were transfected into HTR-8/SVneo trophoblast cells for 24 hours. Results: We observed that IMUPs are mainly expressed in the syncytiotrophoblasts and syncytial knot of placental villi. The expression of IMUP-1 was not differences between normal and preeclamptic placenta tissues. However, IMUP-2 expression was significantly higher in preterm preeclamptic placenta tissues than in normal placenta tissues without labor (p<0.001). Furthermore, we confirmed overexpression of IMUP-2 induced apoptosis in HTR-8/SVneo trophoblast cells through up-regulation of pro-apoptotic proteins. Conclusions: These results suggest that the expression of IMUP-2 is involved in placental development as well as increased IMUP-2 expression is associated with preeclampsia through the inducing of trophoblast apoptosis.

• Journal of the Korean Magnetic Resonance Society
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• v.17 no.1
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• pp.30-39
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• 2013

Immortalization-upregulated protein-1 (IMUP-1) genes have been cloned and are known to be involved in SV40-mediated immortalization. IMUP-1 gene is highly expressed in various cancer cell lines and tumors, suggesting the possibility that they might be involved in tumorigenicity. Previously, there were several problems for overexpression of IMUP-1 in bacterial expression systems including low solubility and aggregation due to unstructured property. To investigate the structural properties, it is necessary to obtain lots of pure and soluble proteins. Accordingly, the co-expression systems of bacterial chaperone proteins, GroEL-GroES, were used to increase solubility of IMUP-1. From the analysis of NMR and CD experiment data, it is suggested that the protein adopt typical the random coil properties in solution.

• Journal of Microbiology and Biotechnology
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• v.34 no.3
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• pp.506-515
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• 2024

Primary human dermal papilla cells (HDPCs) are often preferred in studies on hair growth and regeneration. However, primary HDPCs are limited by their reduced proliferative capacity, decreased hair induction potential, and extended doubling times at higher passages. To overcome these limitations, pTARGET vectors containing human papillomavirus16 (HPV16) E6/E7 oncogenes were transfected into HDPCs and selected using G-148 to generate immortalized cells here. HPV16 E6/E7 oncogenes were efficiently transfected into primary HDPCs. Immortalized HDPC showed higher proliferative activity than primary HDPC, confirming an increased proliferation rate. Expression of p53 and pRb proteins was downregulated by E6 and E7, respectively. E6/E7 expressing HDPC cells revealed that cyclin-dependent kinase (CDK) inhibitor p21 expression was decreased, while cell cycle-related genes and proteins (CDK2 and cyclin E) and E2F family genes were upregulated. Immortalized HDPCs maintained their responsiveness to Wnt/β-catenin pathway and hair follicle formation capability, as indicated by their aggregative properties and stemness. E6/E7 immortalized HDPCs may facilitate in vitro hair growth and regeneration studies.