Our previous study demonstrated that conditional reprogramming (CR) allows the establishment of patient-derived normal and tumor epithelial cell cultures from a variety of tissue types including breast, lung, colon and prostate. Using CR, we have established matched normal and tumor cultures, GUMC-29 and GUMC-30 respectively, from a patient's prostatectomy specimen. These CR cells proliferate indefinitely in vitro and retain stable karyotypes. Most importantly, only tumor-derived CR cells (GUMC-30) produced tumors in xenografted SCID mice, demonstrating maintenance of the critical tumor phenotype. Characterization of cells with DNA fingerprinting demonstrated identical patterns in normal and tumor CR cells as well as in xenografted tumors. By flow cytometry, both normal and tumor CR cells expressed basal, luminal, and stem cell markers, with the majority of the normal and tumor CR cells expressing prostate basal cell markers, CD44 and Trop2, as well as luminal marker, CD13, suggesting a transit-amplifying phenotype. Consistent with this phenotype, real time RT-PCR analyses demonstrated that CR cells predominantly expressed high levels of basal cell markers (KRT5, KRT14 and p63), and low levels of luminal markers. When the CR tumor cells were injected into SCID mice, the expression of luminal markers (AR, NKX3.1) increased significantly, while basal cell markers dramatically decreased. These data suggest that CR cells maintain high levels of proliferation and low levels of differentiation in the presence of feeder cells and ROCK inhibitor, but undergo differentiation once injected into SCID mice. Genomic analyses, including SNP and INDEL, identified genes mutated in tumor cells, including components of apoptosis, cell attachment, and hypoxia pathways. The use of matched patient-derived cells provides a unique in vitro model for studies of early prostate cancer.
Regulation of tyrosine hydroxylase (TH) by intermittent hypoxia (IH) was investigated in rat pheochromocytoma 12 (PC-12) cells by exposing them to alternating cycles of hypoxia (1% O2, 15 s) and normoxia (21% O2, 3 min) for up to 60 cycles; controls were exposed to normoxia for a similar duration. IH exposure increased dopamine content and TH activity by approximately 42 and approximately 56%, respectively. Immunoblot analysis revealed that comparable levels of TH protein were expressed in normoxic and IH cells. Removal of TH-bound catecholamines and in vitro phosphorylation of TH in cell-free extracts by the catalytic subunit of protein kinase A (PKA) increased TH activity in normoxic but not in IH cells, suggesting possible induction of TH phosphorylation and removal of endogenous inhibition of TH by IH. To assess the role of serine phosphorylation in IH-induced TH activation, TH immunoprecipitates and extracts derived from normoxic and IH cells were probed with anti-phosphoserine and anti-phospho-TH (Ser-40) antibody, respectively. Compared with normoxic cells, total serine and Ser-40-specific phosphorylation of TH were increased in IH cells. IH-induced activation of TH and the increase in total serine and Ser-40-specific phosphorylation of TH were inhibited by Ca2+/calmodulin-dependent protein kinase (CaMK) and PKA-specific inhibitors but not by inhibitors of the extracellular signal-regulated protein kinase pathway, suggesting that IH activates TH in PC-12 cells via phosphorylation of serine residues including Ser-40, in part, by CaMK and PKA. Our results also suggest that IH-induced phosphorylation of TH facilitates the removal of endogenous inhibition of TH, leading to increased synthesis of dopamine.
Investigations of mechanisms of human prostate carcinogenesis are limited by the unavailability of a suitable in vitro model system. We have demonstrated that an immortal, but nontumorigenic, human epithelial cell line (267B1) established from fetal prostate tissue can be malignantly transformed by a biological carcinogen, and can serve as a useful model for investigations of the progression steps of carcinogenesis. Activated Ki-ras was introduced into 267B1 cells by infection with the Kirsten murine sarcoma virus. Morphological alterations and anchorage-independent growth were observed; when cells were injected into nude mice, poorly differentiated adenocarcinomas developed. These findings represent the first evidence of malignant transformation of human prostate epithelial cells in culture, and support a role for Ki-ras activation in a multistep process for prostate neoplastic transformation.
With the increase of esthetic demands, most patients want to have restorations which are not only functional but also esthetic. For the esthetic restoration, many ceramic systems have been introduced and applied in dentistry. Among those ceramic restorations, IPS e.max system composed of lithium disilicate glass ceramic is one of the most commonly used systems because it has strength and esthetic characteristics. IPS e.max system is divided into IPS e.max Press and IPS e.max CAD according to the manufacturing methods. IPS e.max Press is fabricated through heat-pressed technique with ceramic ingot, which is very simple. The restorations which are made using IPS e.max system can apply to 3 units restoration for the anterior teeth and premolar, and single posterior tooth restoration. Cementation is one of the most important clinic procedure for the longevity of the restorations. All ceramics are bonded by resin cements, it is classified into three groups including adhesive, self-adhesive, and conventional. Variolink N, which is an adhesive resin cement and manufactured by same company with IPS e.max, is recommended for the bonding of IPS e.max restoration. Conventional and self-adhesive resin cement is also available. The aim of this review article is to provide the understanding of material properties, production procedure and clinical application of IPS e.max system.
This study was carried out in order to offer basic data available for forming the better oral health habit by closely examining a cause for industrial workers' tooth loss. It analyzed correlation in missing teeth with general characteristics, subjective oral health recognition, habit related to oral health, and oral symptom targeting 6,976 of production workers and office workers who work at 49 industrial places where are located in Gumi city of Gyeongsangbuk-do Province from June 1, 2009 to December 31, thereby having obtained the following conclusions. 1. The higher age group led to the higher ratio of missing teeth. The ratio of missing teeth was lower in a case of having been removed plaque than having not been removed plaque. 2. The ratio of missing teeth was high in a case of having dental caries and periodontal disease. The ratio of missing teeth was high in a case of having smoked in the past or in a case of smoking now. 3. The ratio of missing teeth was high in a case of having less toothbrushing frequency and in a case of not using dental floss or interdental brush. 4. The ratio of missing teeth was high when a tooth is throbbing and when tongue and cheek are throbbing in addition to pain and bleeding in gum. 5. The ratio of missing teeth was high in a case of having unpleasant bad breath. The above results are expected to be likely conducive to offering basic data necessary for developing the oral health educational program for industrial places that can promote and maintain workers' oral health, and for improving oral health program and system available for promoting and maintaining workers' oral health hereafter.■ keyword :|Medical Uuse|Missing Teeth|Smoking|Toothbrushing|Worker|
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