scholarly journals Inheritance of gene density–related higher order chromatin arrangements in normal and tumor cell nuclei

2003 ◽  
Vol 162 (5) ◽  
pp. 809-820 ◽  
Author(s):  
Marion Cremer ◽  
Katrin Küpper ◽  
Babett Wagler ◽  
Leah Wizelman ◽  
Johann v. Hase ◽  
...  

A gene density–related difference in the radial arrangement of chromosome territories (CTs) was previously described for human lymphocyte nuclei with gene-poor CT #18 located toward the nuclear periphery and gene-dense CT #19 in the nuclear interior (Croft, J.A., J.M. Bridger, S. Boyle, P. Perry, P. Teague, and W.A. Bickmore. 1999. J. Cell Biol. 145:1119–1131). Here, we analyzed the radial distribution of chromosome 18 and 19 chromatin in six normal cell types and in eight tumor cell lines, some of them with imbalances and rearrangements of the two chromosomes. Our findings demonstrate that a significant difference in the radial distribution of #18 and #19 chromatin is a common feature of higher order chromatin architecture in both normal and malignant cell types. However, in seven of eight tumor cell lines, the difference was less pronounced compared with normal cell nuclei due to a higher fraction of nuclei showing an inverted CT position, i.e., a CT #18 located more internally than a CT #19. This observation emphasizes a partial loss of radial chromatin order in tumor cell nuclei.

1989 ◽  
Vol 9 (11) ◽  
pp. 4722-4730 ◽  
Author(s):  
A L Hiti ◽  
E Bogenmann ◽  
F Gonzales ◽  
P A Jones

Several human rhabdomyosarcoma cell lines, cultured primary tumor explants, and biopsies of tumor and normal skeletal muscle tissue expressed a 2.0-kilobase transcript that hybridized to the mouse muscle determination gene MyoD1. This transcript was found in tumor cell lines and primary explants that developed multinucleated myotubes but was absent in Wilms' tumors or cell lines and primary explants that developed multinucleated myotubes but was absent in Wilms' tumors or cell lines derived from other mesenchymal tumor cell types. Expression of the human homolog of MyoD1 therefore can define a tumor as a rhabdomyosarcoma. Transfection of the mouse MyoD1 gene into the human rhabdomyosarcoma cell line RD increased the ability of the tumor cells to differentiate into multinucleated myotubes and enhanced myosin heavy-chain gene expression but did not decrease tumorigenicity in nude mice.


1994 ◽  
Vol 42 (7) ◽  
pp. 917-929 ◽  
Author(s):  
E Spiess ◽  
A Brüning ◽  
S Gack ◽  
B Ulbricht ◽  
H Spring ◽  
...  

We investigated the appearance and activity of the cysteine proteinase cathepsin B and its physiological inhibitors, stefins A and B, at the cellular level in human tumor cell lines HS-24, derived from a primary lung tumor (squamous cell), and SB-3, derived from a metastasis (lung adenocarcinoma). In addition to cathepsin B, these tumor cells also expressed the immunologically and functionally related cathepsin L, but not cathepsin H. Stefin A was found in HS-24 but not in SB-3 cells; stefin B was found in both cell types. Using a specific fluorogenic cytochemical assay, the intracellular activity of the enzyme was localized and quantified. Thus, the cellular cathepsin B kinetics for the synthetic substrates Z-Arg-Arg-4M beta NA and Z-Val-Lys-Lys-Arg-4M beta NA, its pH dependence and inhibition by E64, stefins A and B, and cystatin C could be determined. From these measurements it appeared that the enzyme exhibited different cleavage rates for these substrates in the different cell types, showed considerable cleavage activity at neutral pH, which was stable under these conditions for extended time periods, and was highly sensitive to the inhibitors E64 and cystatin C but was considerably less sensitive to stefins, particularly stefin A. By conventional light microscopy, confocal laser scanning microscopy, and electron microscopy the enzymatic activity was localized in lysosomes, as expected, but also in the endoplasmic reticulum, nuclear membrane, and plasma membrane. The endoplasmic reticulum is a site at which only pre-mature enzyme forms exist, which are usually not active. The appearance of enzymatic activity at the plasma membrane confirms earlier biochemical and immunofluorescence microscopic investigations. The different sites of localization within the cells make it likely that different forms of the enzyme are expressed simultaneously, which follow alternate ways of processing and sorting. Taken together, the results support an involvement of the enzyme under extracellular conditions in degradative processes.


Rodriguésia ◽  
2020 ◽  
Vol 71 ◽  
Author(s):  
Fernando Duarte Cabral ◽  
Cassia Cristina Fernandes ◽  
Arthur Barcelos Ribeiro ◽  
Iara Squarisi Squarisi ◽  
Denise Crispim Tavares ◽  
...  

Abstract This paper aims to investigate, for the first time, in vitro antitubercular, antileishmanial and antiproliferative activities of essential oils (EOs) from S. odoratissima leaves and flowers - grown in midwestern Brazil - against Mycobacterium tuberculosis, promastigote forms of Leishmania amazonensis and human tumor cell lines. Antimycobacterial activity of EOs was evaluated in terms of the minimal inhibitory concentration (MIC). EOs from leaves and flowers showed to be active antimicrobials against M. tuberculosis, since MIC values were 150 µg/mL and 162.5 µg/mL, respectively. Both EOs exhibited significant activity against promastigote forms of L. amazonensis; IC50 values (50% growth inhibition) were 14.36 ± 2.02 (EOs from leaves) and 19.89 ± 2.66 µg/mL (EOs from flowers). Antiproliferative activity in normal (GM07492A, lung fibroblasts) and tumor (MCF-7, HeLa and M059J) cell lines was performed by the XTT assay; results were expressed as IC50 (50% cell growth inhibition) and the selective index was calculated. IC50 values of EOs from leaves and flowers obtained in normal cell lines for were 502.97 ± 40.33 µg/mL and 370.60 ± 2.01 µg/mL, respectively. Antiproliferative activity was observed against human tumor cell lines, whose IC50 values were significantly lower than those obtained in normal cell lines of MCF-7 cells (367.57 ± 4.46 µg/mL-EOs from leaves and 357.70 ± 1.85 µg/mL-EOs from flowers) and M059J cells (492.53 ± 56.67 µg/mL-EOs from leaves and 324.90 ± 6.72 µg/mL-EOs from flowers), thus, indicating selectivity. These in vitro results showed that EOs from S. odoratissima may be an antimycobacterial, antiparasitic and antitumor agent.


2021 ◽  
Author(s):  
mengyang zhang ◽  
dongxu lin ◽  
changcheng luo ◽  
pengyu wei ◽  
kai cui ◽  
...  

Abstract Background: RNA activation, as a method of regulating gene expression at the transcriptional level, is far less widely used than RNA interference because of the insufficient understanding of the mechanism and the unstable success rate. It is necessary to analyze the failure cases of RNA activation to promote the application of RNA activation. When we validated the saRNAs designed to induce KLK1 expression, we found that saKLK1-374 can up-regulate KLK1 expression in prostate tumor cell lines, but failed in normal prostate cell lines. In addition, we also found that saKLK1-374 inhibited the growth of prostate cancer cells, which seems to be the opposite of the function of KLK1. This article is about experimental research and analysis of these two issues.Methods: To determine whether the phenomenon that the RNA activation of normal cells is difficult to succeed is only valid when the target gene is KLK1, we used p21WAF1/CIP1 as the target gene to perform RNA activation experiments in normal prostate cells and prostate cancer cells. Next, to determine whether the above phenomenon exists in other tissues, we also performed RNA activation experiments with KLK1 and p21WAF1/CIP1 as target genes in normal cell lines and tumor cell lines derived from the bladder. We have also extended the time from transfection to the detection of target gene expression to evaluate whether a longer saRNA action time can change the phenomenon that saRNA fails to up-regulate target gene expression in normal cells. In terms of mechanism research, we used fluorescently labeled dsRNA to evaluate the transfection efficiency, and also detected the expression of Ago2 and IPO8 proteins. In another issue of saKLK1-374 inhibiting prostate cancer cells, we tested the ROS content and apoptosis levels of prostate cancer cells after saKLK1-374 transfection. We used recombinant KLK1 protein to directly interfere with prostate cancer cells as a positive control for KLK1 function research. In turn, we also used siRNA to inhibit the expression of KLK1 in prostate cancer cells to compare the growth of prostate cancer cells when KLK1 mRNA was up-regulated and reduced.Results: The p21WAF1/CIP1 gene could be significantly upregulated by saRNA in prostate cancer cell lines, but not in normal prostate cell lines. The expression of KLK1 in bladder-derived cell lines was extremely low and could not be induced by saRNA. The p21WAF1/CIP1 gene could be up-regulated by saRNA to a higher extent in bladder cancer cell lines, while it was up-regulated by saRNA in normal urothelial cell line to a lower extent. Prolonging the action time of saRNA could not change that saRNA failed to induce the expression of target genes in normal cell lines. Compared with tumor cell lines, normal cell lines had lower transfection efficiency or lower expression of Ago2 and IPO8. After being transfected with saKLK1-374, prostate cancer cells had increased ROS and increased levels of apoptosis. The recombinant KLK1 protein did not increase ROS in prostate cancer cells, nor did it inhibit their growth. Even though saKLK1-374 up-regulated the expression of KLK1 in prostate cancer cells, siRNA still suppressed the expression of KLK1 below the baseline level, and in this case, the growth of prostate cancer cells was still at a suppressed level.Conclusion: Normal cell lines may be more difficult to be successfully induced target gene expression than tumor cells due to low transfection efficiency or low Ago2 and IPO8 expression. In addition, although saKLK1-374 is designed to up-regulate the expression of KLK1, the reason that it inhibits the proliferation of prostate cancer cells is irrelevant to the up-regulated expression of KLK1.


2008 ◽  
pp. 935-945
Author(s):  
M Orečná ◽  
R Hafko ◽  
Z Bačová ◽  
J Podskočová ◽  
D Chorvát ◽  
...  

Objective of this study was to characterize osmotically-induced insulin secretion in two tumor cell lines. We compared response of freshly isolated rat pancreatic islets and INS-1 and INS-1E tumor cell lines to high glucose, 30 % hypotonic medium and 20 % hypertonic medium. In Ca(2+)-containing medium glucose induced insulin release in all three cell types. Hypotonicity induced insulin secretion from islets and INS-1 cells but not from INS-1E cells, in which secretion was inhibited despite similar increase in cell volume in both cell types. GdCl(3) (100 micromol/l) did not affect insulin response from INS-1E cells to hypotonic challenge. Hypertonic medium inhibited glucose-induced insulin secretion from islets but not from tumor cells. Noradrenaline (1 micromol/l) inhibited glucose-induced but not swelling-induced insulin secretion from INS-1 cells. Surprisingly, perifusion with Ca(2+)-depleted medium showed distinct secretory response of INS-1E cells to hypotonicity while that of INS-1 cells was partially inhibited. Functioning glucose-induced insulin secretion is not sufficient prerequisite for hypotonicity-induced response in INS-1E cells suggesting that swelling-induced exocytosis is not essential step in the mechanism mediating glucose-induced insulin secretion. Both cell lines are resistant to inhibitory effect of hyperosmolarity on glucose-induced insulin secretion. Response of INS-1E cells to hypotonicity is inhibited by the presence of Ca(2+) in medium.


1989 ◽  
Vol 9 (11) ◽  
pp. 4722-4730
Author(s):  
A L Hiti ◽  
E Bogenmann ◽  
F Gonzales ◽  
P A Jones

Several human rhabdomyosarcoma cell lines, cultured primary tumor explants, and biopsies of tumor and normal skeletal muscle tissue expressed a 2.0-kilobase transcript that hybridized to the mouse muscle determination gene MyoD1. This transcript was found in tumor cell lines and primary explants that developed multinucleated myotubes but was absent in Wilms' tumors or cell lines and primary explants that developed multinucleated myotubes but was absent in Wilms' tumors or cell lines derived from other mesenchymal tumor cell types. Expression of the human homolog of MyoD1 therefore can define a tumor as a rhabdomyosarcoma. Transfection of the mouse MyoD1 gene into the human rhabdomyosarcoma cell line RD increased the ability of the tumor cells to differentiate into multinucleated myotubes and enhanced myosin heavy-chain gene expression but did not decrease tumorigenicity in nude mice.


Author(s):  
Iara Squarisi ◽  
Karoline Soares De Freitas ◽  
Danieli Cristina Lemes ◽  
Gari Vidal Ccana Ccapatinta ◽  
Jennyfer Andrea Aldana Mejía ◽  
...  

Abstract. Squarisi IS, De Freitas KS, Lemes DC, Ccana-Ccapatinta GV, Mejia JAA, Bastos JK, Veneziani RCS, Ambrosio SR, Tavares DC. 2018. Evaluation of the antiproliferative activity of red propolis hydroalcoholic extract and its fractions obtained by partition. Biofarmasi J Nat Prod Biochem 18: 66-69. The present study aimed to evaluate the cytotoxicity of red propolis hydroalcoholic extract (RPHE) and its fractions obtained by partition, hexanes (HF), dichloromethane (DF), ethyl acetate (AF) and n-butanol (BF), on tumor and non-tumor cell lines. For this purpose, the XTT colorimetric assay was performed on human lung fibroblasts (GM07492A, non-tumor cell), breast adenocarcinoma (MCF-7), glioblastoma (U343) and cervix adenocarcinoma (HeLa) cells. The results showed that RPHE, HF and DF presented not only cytotoxic potential to all tumor cell lines but also to normal cell line, indicating selectivity absence. HF presented the lowest IC50 (half minimal inhibitory concentration; 33.8-133.3 µg/mL), with significant difference from those observed for RPHE (137.0-262.7 µg/mL). BF and AF revealed an IC50 which was higher than 1250 µg/mL in all cell lines. The results showed that red propolis has substances with antiproliferative activity, indicating that its hexanes fraction may have substances with antitumor potential.


2013 ◽  
Vol 27 (S1) ◽  
Author(s):  
Fang Jin ◽  
Toby Terwilliger ◽  
Jessica Bosch ◽  
Robert W. O'Donnell

2020 ◽  
Vol 4 (1) ◽  
pp. 13-20
Author(s):  
Reshna K. Ahmad ◽  
Kawa F. Dizaye ◽  
Asaad A. AL-Asady

Reports indicated that extract Pistacia khinjuk has anti-inflammatory, antipyretic, antibacterial, and antiviral, in treating of diarrhea and throat infections and has hepatoprotective effects against acetaminophen and carbon tetrachloride. This study was undertaken to investigate the possible cytotoxic effects of methanolic and aqueous seeds extract of P. khinjuk on different tumors (rhabdomyosarcoma [RD] and murine mammary adenocarcinoma [Ahmed-Mohammed-Nahi-2003 (AMN-3)]) and normal cell lines (murine fibroblast) and its mitogenic effects on blood lymphocytes. The cytotoxic effects of P. khinjuk seed extracts were evaluated on two tumor cell lines, RD and murine mammary adenocarcinoma (AMN-3) and one normal cell line, murine fibroblast (L20B). Moreover, the mitogenic effects of the plant extract were studied, on human blood lymphocytes. Both methanolic and aqueous seed extracts of P. khinjuk significantly induced tumor cell lines and the normal cell line proliferation, especially in highest concentrations. The results show that the extracts induced significant increases in human blood lymphocyte proliferation at 72 h. This activity of plant extracts recommends it as a good mitogenic agent in researches; in conclusion, seed extracts of P. khinjuk induced proliferation of all tested cell lines. High concentrations of both aqueous and methanolic seed extracts of P. khinjuk showed mitogenic effects.


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