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101.
102.
Caveolin‐1 down‐regulation is required for Wnt5a‐Frizzled 2 signalling in Ha‐RasV12‐induced cell transformation 下载免费PDF全文
Hsiu‐Kuan Lin Hsi‐Hui Lin Yu‐Wei Chiou Ching‐Lung Wu Wen‐Tai Chiu Ming‐Jer Tang 《Journal of cellular and molecular medicine》2018,22(5):2631-2643
Caveolin‐1 (Cav1) is down‐regulated during MK4 (MDCK cells harbouring inducible Ha‐RasV12 gene) transformation by Ha‐RasV12. Cav1 overexpression abrogates the Ha‐RasV12‐driven transformation of MK4 cells; however, the targeted down‐regulation of Cav1 is not sufficient to mimic this transformation. Cav1‐silenced cells, including MK4/shCav1 cells and MDCK/shCav1 cells, showed an increased cell area and discontinuous junction‐related proteins staining. Cellular and mechanical transformations were completed when MDCK/shCav1 cells were treated with medium conditioned by MK4 cells treated with IPTG (MK4+I‐CM) but not with medium conditioned by MK4 cells. Nanoparticle tracking analysis showed that Ha‐RasV12‐inducing MK4 cells increased exosome‐like microvesicles release compared with their normal counterparts. The cellular and mechanical transformation activities of MK4+I‐CM were abolished after heat treatment and exosome depletion and were copied by exosomes derived from MK4+I‐CM (MK4+I‐EXs). Wnt5a, a downstream product of Ha‐RasV12, was markedly secreted by MK4+I‐CM and MK4+I‐EXs. Suppression of Wnt5a expression and secretion using the porcupine inhibitor C59 or Wnt5a siRNA inhibited the Ha‐RasV12‐ and MK4+I‐CM‐induced transformation of MK4 cells and MDCK/shCav1 cells, respectively. Cav1 down‐regulation, either by Ha‐RasV12 or targeted shRNA, increased frizzled‐2 (Fzd2) protein levels without affecting its mRNA levels, suggesting a novel role of Cav1 in negatively regulating Fzd2 expression. Additionally, silencing Cav1 facilitated the internalization of MK4+I‐EXs in MDCK cells. These data suggest that Cav1‐dependent repression of Fzd2 and exosome uptake is potentially relevant to its antitransformation activity, which hinders the activation of Ha‐RasV12‐Wnt5a‐Stat3 pathway. Altogether, these results suggest that both decreasing Cav1 and increasing exosomal Wnt5a must be implemented during Ha‐RasV12‐driven cell transformation. 相似文献
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Investigating the mincing method for isolation of adipose-derived stem cells from pregnant women fat
Yuan-Sheng Li Pao-Jen Chen Li-Wei Wu Pei-Wen Chou Li-Yi Sun Tzyy-Wen Chiou 《Cytotechnology》2018,70(1):55-66
The success of stem cell application in regenerative medicine, usually require a stable source of stem or progenitor cells. Fat tissue represents a good source of stem cells because it is rich in stem cells and there are fewer ethical issues related to the use of such stem cells, unlike embryonic stem cells. Therefore, there has been increased interest in adipose-derived stem cells (ADSCs) for tissue engineering applications. Here, we aim to provide an easy processing method for isolating adult stem cells from human adipose tissue harvested from the subcutaneous fat of the abdominal wall during gynecologic surgery. We used a homogenizer to mince fat and compared the results with those obtained from the traditional cut method involving a sterile scalpel and forceps. Our results showed that our method provides another stable and quality source of stem cells that could be used in cases with a large quantity of fat. Furthermore, we found that pregnancy adipose-derived stem cells (P-ADSCs) could be maintained in vitro for extended periods with a stable population doubling and low senescence levels. P-ADSCs could also differentiate in vitro into adipogenic, osteogenic, chondrogenic, and insulin-producing cells in the presence of lineage-specific induction factors. In conclusion, like human lipoaspirates, adipose tissues obtained from pregnant women contain multipotent cells with better proliferation and showed great promise for use in both stem cell banking studies as well as in stem cell therapy. 相似文献
105.
Jiahn-Shing Lee Jiahn-Haur Liao Shih-Hsiung Wu Shyh-Horng Chiou 《Journal of Protein Chemistry》1997,16(4):283-289
-Crystallin, a major protein of the eye lens, is known to have chaperone activity in preventing heat-induced aggregation of enzymes and other crystallins. In this study, we investigate the ability of -crystallin to inhibit UV-light-induced aggregation of other lens proteins and the effect of exposure of -crystallin to UV irradiation on its chaperone activity. The chaperone activities of -crystallin preincubated at different temperatures were found to be different and could be correlated with its change in quaternary structure as determined by the fluorescence probe ANS (8-anilo-1-naphthalene sulfonate). -Crystallin can inhibit the aggregation of -crystallin from UV irradiation at room temperature, and the preheated -crystallins provide more protection than the native one. Upon irradiation by UV light, -crystallin gradually lost its ability to protect -crystallin against thermal aggregation. The loss of the chaperone efficacy of -crystallin to protect other lens proteins may shed light on human cataract formation induced by long-term exposure to UV irradiation. 相似文献
106.
Antheridiogen chemicals secreted by living fern gametophytes have been shown to influence production of male gametangia and thus mating systems in a large number of terrestrial fern species. Antheridiogens have not previously been thought to be prevalent in the Polypodiaceae, a large family composed mostly of tropical epiphytes. This study presents bioassay methods more sensitive than previously used to detect antheridiogen and demonstrates that antheridiogens are also operative in the Polypodiaceae and in epiphytic species. Seven species in six genera (Campyloneurum angustifolium, C. phyllitidis, Lepisorus thunbergianus, Microgramma heterophylla, Phlebodium aureum, Phymatosorus scolopendria, and Polypodium pellucidum) were tested for the presence of an antheridiogen system. All species tested except P. aureum were induced to produce antheridia precociously by their own antheridiogen and by that of Pteridium aquilinum (APt). Phlebodium aureum responded to APt and promoted antheridium formation in Onoclea sensibilis but did not respond to its own antheridiogen. Spores of all species except P. aureum were induced to germinate in darkness by antheridiogen of the same species and by APt and to form antheridia in the dark, further enhancing the possibility of intergametophytic mating. 相似文献
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Due to their bio-compatibility and high porosity, cellulose foam and polyurethane foam (PUF) were selected as the media to immobilize insect cells for recombinant baculovirus expression. In packed beds, entrapment efficiency and axial cell distribution are two prime important parameters associated with reactor performance. These two parameters were found to be affected by initial cell loading density and liquid circulation rate. Based on the entrapment kinetic model, the entrapment parameters as K 0, σ m could be evaluated. The establishment of the correlation between K 0, σ m and Reynolds number, N Re, described in this study provided convenient tools to achieve an efficient cell entrapment in the packed bed. The appropriated circulation velocity for uniform cell distribution was also determined. 相似文献
109.
L. M. Chiou M. C. Chung† P. G. Tung‡ T. Hsu§ J. S. Yang¶ 《Journal of fish biology》2004,65(6):1614-1621
An immuno‐probe against a glycoprotein in the egg chorion was developed for egg identification. The 97 kD glycoprotein in the chorion of unfertilized eggs of Epinephelus malabaricus was isolated and separated by SDS‐PAGE as an antigen to induce antibody from rabbit. The reactivity of the antibody as the immuno‐probe to E. malabaricus eggs was significantly positive, and was specific in that it did not react with the eggs of other fish species. The immuno‐probe should be useful in identifying the eggs of E. malabaricus among mixed egg populations. 相似文献
110.