scholarly journals Changes in mRNA levels of the sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase isoforms in the rat soleus muscle regenerating from notexin-induced necrosis

1996 ◽  
Vol 320 (1) ◽  
pp. 107-113 ◽  
Author(s):  
Ernö ZÁDOR ◽  
Luca MENDLER ◽  
Mark VER HEYEN ◽  
László DUX ◽  
Frank WUYTACK

The relative mRNA levels corresponding to the different sarcoplasmic/endoplasmic-reticulum Ca2+-ATPase isoforms (SERCA1a, SERCA1b, SERCA2a, SERCA2b and SERCA3) were measured by reverse transcriptase–PCR in rat soleus muscles regenerating after notexin-induced necrosis. The succession of appearance of the different types of SERCA mRNA species in regenerating muscle largely recapitulates those observed during normal ontogenesis. The mRNA levels of the muscle-specific isoforms SERCA1a and SERCA2a became very low on the first and third days after injection of the snake venom. It was only on the fifth day of regeneration that the mRNA of the neonatal variant of the fast-twitch skeletal SERCA1b isoform began to rise, well before the other SERCA transcripts. At 7 and 10 days, i.e. at a time when the new myofibres normally become re-innervated, the mRNA level of SERCA1a and SERCA2a increased markedly, but the fast-twitch skeletal SERCA1a isoform was still the most prominent. On day 21, in the advanced stage of regeneration, a switch in the relative expression levels of SERCA1a and SERCA2a mRNA was observed and the ratio of both isoforms became similar to that found in the normal soleus muscles. This was followed by a decline in the level of all SERCA mRNA species, so that on day 28 the levels of the sarcoplasmic/endoplasmatic-reticulum Ca2+-pump RNAs was again lower but their ratio remained similar to that of the untreated control soleus.

2021 ◽  
Vol 21 ◽  
Author(s):  
Reem Mebed ◽  
Yasser BM Ali ◽  
Nahla Shehata ◽  
Nadia El-Guendy ◽  
Nahla Gamal ◽  
...  

Background: Bevacizumab (Bev) resistance is hypothesized to be overcome by combination with inhibitors of other signalling pathways. Objective: We aimed to study the effect of combining Bev with knocked down β-catenin (Bev-β-cat-siRNA) on the expression of VEGF-A, Slug, NFКB and its two target genes c-Flip and FasR in HepG2. Expression of VEGF-A and Slug was also studied in Caco-2 cells. Methods: Cultured cells were divided into six groups 1) cells treated with Bev only 2) cells treated with β-catenin-siRNA 3) cells treated with Bev-β-cat-siRNA 4) cells treated with negative control 5) cells treated with Bev-negative control and untreated cells. Expressions were assessed using qPCR and western blotting. Results: Bev-β-cat-siRNA significantly reduced the mRNA level of VEGF-A, which was initially increased in response to Bev alone in HepG2 but not in Caco-2. Additionally, Bev-β-cat-siRNA significantly decreased Slug mRNA level compared to Bev only treated HepG2 cells. In contrast, VEGF-A and Slug mRNA levels in Bev only group were remarkably lower than Bev-β-cat-siRNA in Caco-2 cells. Distinct β-catenin and Slug protein expressions were noticed in HepG2 and Caco-2 cells. On the other hand, Bev-β-cat-siRNA remarkably reduced the level of NFКB, FasR and c-Flip compared to Bev only treated HepG2 cells although the difference was not statistically significant. Conclusion: We conclude that, combining Bevacizumab with knocked down β-catenin reduce the expression of VEGF-A and Slug in HepG2 but not in Caco-2 cells.


1997 ◽  
Vol 17 (6) ◽  
pp. 595-601 ◽  
Author(s):  
Isao Ebihara ◽  
Tsukasa Nakamura ◽  
Toshimasa Takahashi ◽  
Yasuhiko Tomino ◽  
Noriaki Shimada ◽  
...  

Objective To compare plasma endothelin (ET)-1 level and ET-1 mRNA level in peripheral blood monocytes of patients undergoing hemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD). Design Endothelin-1 mRNA level in peripheral blood monocytes and plasma ET -1 level were studied in 30 HD patients, 15 CAPD patients, 20 chronic renal failure patients not being dialyzed, and 20 normal healthy controls. Hemodialysis patients were dialyzed three times per week with a bicarbonate dialysate. Different types of dialyzer membrane, viz., cellulose triacetate, cuprophane, poly-sulfone, polyacrylonitrile, and ethylenevinylalcohol were used in 8,6,6,5, and 5 patients, respectively. Continuous ambulatory peritoneal dialysis patients were dialyzed with four daily exchanges of a 2-L dialysate containing glucose at a concentration of 1.5% to 2.5%. Results Higher levels of ET -1 mRNA in monocytes were observed in HD patients than in CAPD patients (p < 0.01), chronic renal failure patients (p < 0.01), or normal healthy controls (p < 0.001). The level of ET -1 mRNA in monocytes at the end of HD was not significantly higher than that at the start of HD. ln addition, these mRNA levels in HD patients showed littledifference with different types of dialysis membrane. Plasma ET -1 level in HD patients (10.2 ± 2.4 pg/mL) was also higher than that in CAPD patients (7.8 ± 1.6 pg/mL, p < 0.01), in chronic renal failure patients (4.8 ± 1.2 pg/mL, p < 0.01), or in normal controls (2.6 ± 0.8 pg/mL, p < 0.001). Conclusion Dialysis itself did not significantly affect ET -1 mRNA levels in monocytes. Chronic stimulation of peripheral blood monocytes may be associated with higher levels of ET -1 mRNA and plasma ET -1 in HD patients than in CAPD patients.


2003 ◽  
Vol 77 (14) ◽  
pp. 7936-7944 ◽  
Author(s):  
Kristina Löhr ◽  
Oliver Hartmann ◽  
Helmut Schäfer ◽  
Matthias Dobbelstein

ABSTRACT During infection with adenovirus, massive changes in the transcription of virus genes are observed, suggesting that the expression of cellular genes may also be modulated. To characterize the levels of cellular RNA species in infected cells, cDNA arrays were screened 24 h after infection of HeLa cells with wild-type adenovirus type 5, strain dl309. Despite complete transduction of the cells, fewer than 20 cellular genes (out of 4,600 analyzed and 1,200 found detectable and expressed above background) were altered more than threefold in their corresponding RNA levels compared to mock-infected cells. In particular, the expression of the myc oncogene was reduced at the mRNA level. This reduction was dependent on the replication of virus DNA and partially dependent on the presence of the adenovirus gene products E1B-55 kDa and E4orf6, but not E4orf3. On the other hand, MYC protein had an increased half-life in infected cells, resulting in roughly constant steady-state protein levels. The adenovirus E1A gene product is necessary and sufficient to stabilize MYC. Overexpressed MYC inhibited adenovirus replication and the proper formation of the virus replication centers. We conclude that adenovirus infection leads to the stabilization of MYC, perhaps as a side effect of E1A activities. On the other hand, myc mRNA levels are negatively regulated during adenovirus infection, and this may avoid the detrimental effect of excessive MYC on adenovirus replication.


1993 ◽  
Vol 265 (6) ◽  
pp. L636-L643 ◽  
Author(s):  
B. Maitre ◽  
L. Jornot ◽  
A. F. Junod

We examined the effects of inhibition of Cu,Zn superoxide dismutase (Cu,Zn SOD) and catalase (Cat) activities on the steady-state mRNA levels of the three major antioxidant enzymes [Cu,Zn SOD, Cat, and glutathione peroxidase (GP)] in human umbilical vein endothelial cells under normoxia and hyperoxia. Inhibition of Cat activity by aminotriazole was not associated with alteration of the other antioxidant enzymes or with potentiation of cell injury. On the other hand, inhibition of Cu,Zn SOD activity by N-N'-diethyl-dithiocarbamate (DDC) was associated with an increase in Cu,Zn SOD mRNA level and a decrease in Cat and GP mRNA level. The combination of DDC and hyperoxia treatments was associated with an additive effect on Cu,Zn SOD message. We propose that these coordinate mRNA changes might be an adaptation to the oxidative environment. This proposal supports the concept that the intracellular O2 metabolites themselves could be the signals that trigger the antioxidant enzymes gene expression.


2021 ◽  
Vol 49 (12) ◽  
pp. 030006052110653
Author(s):  
Seung Geun Yeo ◽  
Sung Jong Lee ◽  
Ji Woo Lee ◽  
Sujung Oh ◽  
Dong Choon Park

Objective To compare the levels of endoplasmic reticulum (ER) stress-associated mRNAs and the clinical characteristics of patients with endometriosis or gynaecological cancer. Methods This prospective study obtained intraperitoneal fluid samples from female patients that underwent surgery. The levels of ER stress mRNAs in the peritoneal fluid, including C/EBP-homologous protein (CHOP), X-box binding protein 1 (sXBP1), activating transcription factor 6 (ATF6), immunoglobulin heavy chain-binding protein (BiP), inositol-requiring enzyme 1α (IRE1α) and protein kinase RNA-like endoplasmic reticulum kinase (PERK), were measured using real-time reverse transcription–polymerase chain reaction in patients with benign disease without endometriosis (control group), with endometriosis or with gynaecological cancer. Results This study enrolled 126 patients: 46 control patients; 47 with endometriosis; and 33 with cancer. The levels of CHOP and BiP mRNA were significantly higher in the control group compared with the cancer group. Levels of sXBP1 and ATF6 mRNA were significantly higher in the cancer group than in the control and endometriosis groups. In the endometriosis group, ATF6 mRNA level was inversely correlated with age and positively correlated with serum cancer antigen 125 levels; and ATF6 and PERK mRNA levels were inversely correlated with parity. Conclusion The levels of ER stress-related mRNAs were related to the pathogenesis of endometriosis and gynaecological cancers.


2021 ◽  
Author(s):  
Ting Ye ◽  
Wei-ting Xuan ◽  
Peng Zhou ◽  
Nan Shao ◽  
Hang Song ◽  
...  

Abstract Background Huangdi Anxiao (HDAX) is mainly used to treat diabetes and its complications for many years and has a remarkable curative effect. However, the improvement effect of HDAX in the diabetic cognitive dysfunction (DCD) model and the related mechanism is not clear. This study was aimed to explore the neuroprotective effects of HDAX and its possible mechanisms in DCD. Methods A DCD cell model was established by high glucose-induced PC12 cells, and the effect of HDAX on the cell viability was examined by MTT. Additionally, the expression of relevant genes and proteins in the apoptosis pathway of endoplasmic reticulum (ER) stress was detected. Results The results showed that HDAX increased cell viability, reduced GRP78, CHOP, Bax, procaspase-12, procaspase-9, procaspase-3 mRNA levels and GRP78, CHOP, Bax, Caspase-12, Caspase-9, Caspase-3 protein expressions, and decreased Bcl-2 mRNA level and protein expression. Conclusions These results suggested that HDAX had neuroprotective effects in the DCD cell model, which may be associated with the inhibition of the apoptosis pathway of ER stress.


Author(s):  
R. N. Tomas

Peridinium balticum appears to be unusual among the dinoflagellates in that it possesses two DNA-containing structures as determined by histochemical techniques. Ultrastructurally, the two dissimilar nuclei are contained within different protoplasts; one of the nuclei is characteristically dinophycean in nature, while the other is characteristically eucaryotic. The chloroplasts observed within P. balticum are intrinsic to an eucaryotic photosynthetic endosymbiont and not to the dinoflagellate. These organelles are surrounded by outpocketings of endoplasmic reticulum which are continuous with the eucaryotic nuclear envelope and are characterized by thylakoids composed of three apposed lamellae. Girdle lamellae and membranebounded interlamellar pyrenoids are also present. Only the plasmalemma of the endosymbiont segregates its protoplast from that of the dinophycean cytoplasm. The exact nature of this symbiotic relationship is at present not known.


Author(s):  
S.R. Allegra

The respective roles of the ribo somes, endoplasmic reticulum, Golgi apparatus and perhaps nucleus in the synthesis and maturation of melanosomes is still the subject of some controversy. While the early melanosomes (premelanosomes) have been frequently demonstrated to originate as Golgi vesicles, it is undeniable that these structures can be formed in cells in which Golgi system is not found. This report was prompted by the findings in an essentially amelanotic human cellular blue nevus (melanocytoma) of two distinct lines of melanocytes one of which was devoid of any trace of Golgi apparatus while the other had normal complement of this organelle.


2018 ◽  
Vol 18 (3) ◽  
pp. 287-294 ◽  
Author(s):  
Gustavo Alencastro Veiga Cruzeiro ◽  
Maristella Bergamo dos Reis ◽  
Vanessa Silva Silveira ◽  
Regia Caroline Peixoto Lira ◽  
Carlos Gilberto Carlotti Jr ◽  
...  

Background: Genetic and epigenetic modifications are closely related to tumor initiation and progression and can provide guidance for understanding tumor functioning, potentially leading to the discovery of new therapies. Studies have associated hypoxia-related genes to tumor progression and chemo/radioresistance in brain tumors. Information on the expression profile of hypoxiarelated genes in pediatric medulloblastoma, although scarce, may reveal relevant information that could support treatment decisions. Objective: Our study focused on evaluation the of CA9, CA12, HIF1A, EPAS1, SCL2A1 and VEGF genes in 41 pediatric fresh-frozen medulloblastoma sample. Additionally, we analyzed the effect of hypoxia and normoxia in the pediatric medulloblastoma cell-line UW402. Furthermore, we assessed the effects of HIF1A knockdown in cell-proliferation and methylation levels of genes related to hypoxia, apoptosis and autophagy. Method: qPCR was performed to evaluate mRNA levels, and Western blot to confirm HIF1A silencing in both patient samples and cell line. Pyrosequencing was performed to asses the methylation levels after HIF1A knockdown in the UW402 cell line. Results: A higher HIF1A mRNA level was observed in MB patients when compared to the cerebellum (non-tumor match). In UW402 MB cell-line, chemically induced hypoxic resulted in an increase of mRNA levels of HIF1A, VEGF, SCL2A1 and CA9 genes. Additionally, HIF1A knockdown induced a decrease in the expression of hypoxia related genes and a decrease of 30% in cell proliferation was also observed. Also, a significant increase in the methylation of ATG16L1 promoter and decrease in the methylation of EPAS1 promoter were observed after HIF1A knockdown. Conclusion: HIF1A knockdown in medulloblastoma cells lead to decreased cellular proliferation, suggesting that HIF1A can be a potential therapeutic target to be explored in the medulloblastoma. However, the mechanisms behind HIF1A protein stabilization and function are very complex and more data need to be generated to potentially use HIF1A as a therapeutical target.


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