Biochemical Study of Nuclei Isolated From Normal Lung and Lung Tumors. I. Isolation of Nuclei and Characterization of Nuclear RNA2

2020 ◽  
Vol 41 (11) ◽  
pp. 1518-1528
Author(s):  
Sreekanth Chanickal Narayanapillai ◽  
Yong Hwan Han ◽  
Jung Min Song ◽  
Manaye Ebabu Kebede ◽  
Pramod Upadhyaya ◽  
...  

Abstract Chronic obstructive pulmonary disease (COPD) is a significant risk factor for lung cancer. One potential mechanism through which COPD contributes to lung cancer development could be through generation of an immunosuppressive microenvironment that allows tumor formation and progression. In this study, we compared the status of immune cells and immune checkpoint proteins in lung tumors induced by the tobacco smoke carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) or NNK + lipopolysaccharide (LPS), a model for COPD-associated lung tumors. Compared with NNK-induced lung tumors, NNK+LPS-induced lung tumors exhibited an immunosuppressive microenvironment characterized by higher relative abundances of PD-1+ tumor-associated macrophages, PD-L1+ tumor cells, PD-1+ CD4 and CD8 T lymphocytes and FOXP3+ CD4 and CD8 T lymphocytes. Also, these markers were more abundant in the tumor tissue than in the surrounding ‘normal’ lung tissue of NNK+LPS-induced lung tumors. PD-L1 expression in lung tumors was associated with IFNγ/STAT1/STAT3 signaling axis. In cell line models, PD-L1 expression was found to be significantly enhanced in phorbol-12-myristate 13-acetate activated THP-1 human monocytes (macrophages) treated with LPS or incubated in conditioned media (CM) generated by non-small cell lung cancer (NSCLC) cells. Similarly, when NSCLC cells were incubated in CM generated by activated THP-1 cells, PD-L1 expression was upregulated in EGFR- and ERK-dependent manner. Overall, our observations indicate that COPD-like chronic inflammation creates a favorable immunosuppressive microenvironment for tumor development and COPD-associated lung tumors might show a better response to immune checkpoint therapies.


Molecules ◽  
2020 ◽  
Vol 25 (17) ◽  
pp. 3823 ◽  
Author(s):  
Madumani Amararathna ◽  
David W. Hoskin ◽  
H. P. Vasantha Rupasinghe

In our previous study, we demonstrated that cyanidin-3-O-glucoside (C3G)-rich haskap (Lonicera caerulea L.) berry extracts can attenuate the carcinogen-induced DNA damage in normal lung epithelial cells in vitro. Here, the efficacy of lyophilized powder of whole haskap berry (C3G-HB) in lowering tobacco-specific nitrosamine, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, (NNK)-induced lung tumorigenesis in A/JCr mice was investigated. Three weeks after daily oral administration of C3G-HB (6 mg of C3G in 0.2 g of C3G-HB/mouse/day), lung tumors were initiated by a single intraperitoneal injection of NNK. Dietary C3G-HB supplementation was continued, and 22 weeks later, mice were euthanized. Lung tumors were visualized through positron emission tomography (PET) and magnetic resonance imaging (MRI) 19 weeks after NNK injection. Dietary supplementation of C3G-HB significantly reduced the NNK-induced lung tumor multiplicity and tumor area but did not affect tumor incidence. Immunohistochemical analysis showed reduced expression of proliferative cell nuclear antigen (PCNA) and Ki-67 in lung tissues. Therefore, C3G-HB has the potential to reduce the lung tumorigenesis, and to be used as a source for developing dietary supplements or nutraceuticals for reducing the risk of lung cancer among high-risk populations.


2014 ◽  
Vol 55 (5) ◽  
pp. 480-487 ◽  
Author(s):  
Laura E. Barrett ◽  
Rachel E. Pollard ◽  
Allison Zwingenberger ◽  
Alexandra Zierenberg-Ripoll ◽  
Katherine A. Skorupski

1987 ◽  
Vol 65 (2) ◽  
pp. 173-182 ◽  
Author(s):  
Michael Goldenthal ◽  
James T. Nishiura

A DNA-dependent RNA polymerase was solubilized from sucrose gradient isolated, DNase-treated mitochrondria of Drosophila melanogaster. The isolated mitochondria were not detectably contaminated with nuclear DNA as shown by CsCl gradient centrifugation and polylysine Kieselguhr chromatography. The detergent-solubilized RNA polymerase was sensitive to rifampicin, resistant to α-amanitin, had an apparent molecular mass of about 60 kilodaltons, and displayed a tendency to aggregate, both in crude extracts or when purified. The mitochondrial RNA polymerase could be distinguished from nuclear RNA polymerases on the basis of size, salt optima, rifampicin sensitivity, and α-amanitin resistance.


2001 ◽  
Vol 113 (1) ◽  
pp. 109-115 ◽  
Author(s):  
Appolinaire Djikeng ◽  
Ludmila Ferreira ◽  
Maximiliano D'Angelo ◽  
Pavel Dolezal ◽  
Tracey Lamb ◽  
...  

OENO One ◽  
1998 ◽  
Vol 32 (1) ◽  
pp. 17
Author(s):  
Ferjani Ben Abdallah ◽  
Farhat Chibani ◽  
Asma Fnayou ◽  
Abdelwahed Ghorbel ◽  
Jean-Michel Boursiquot

<p style="text-align: justify;">61 tunisian autochton grapevine varieties have been collected for biochemical identification. Isozymes analysis with starch gel electrophoresis technique was used to confirn or to cancel random denominations awarded to the majority of these local varieties. In our conditions, concentrated plant extracts were obtained from vigorous donnant canes newly cut off from selected mother plants during automn. These allowed us to dispose of rigorously interpretable isozyme banding patterns of GPI and PGM systems and to overcome difficulties often related to the use of PGM system. The study of GPII and PGM enzyme systems allowed us to classify the autochton accessions into 16 different groups from which 5 groups containing only 2 or 3 varieties.</p><p style="text-align: justify;">On the other hand, the study of AAT and peroxydase enzyme systems has shown stable and legible isozyme banding patterns allowing to discriminate between equivalent accessions such as Sakasly and Kahli (two black local vines very similar), 3 varieties of Bidh Hamem (Bidh Hamem, Bidh Hamem Rafraf and Bidh Hamem Sfax), and 2 varieties of Bezzoul Kelba Bidha (Sfax and Gabes). In addition, certain varieties having for longtime the same denominations were characterized. A case of point the 4 varieties Khalt meaning mixture (Bouchemma, Abiedh, Mdaouer and Souche 1) and the 3 varieties of Arich (Ahmar, Dressée, and Jerba) were proved to be completely different from each other. In the same way, Bezzoul Khadem has been differed from Hemri variety. The complementary use of berry colour allowed to discriminate between Saouadi, Khdhiri and Jebbi varieties and to subdivise the remainig groups into sub-groups.</p><p style="text-align: justify;">The study of GPI, PGM, AAT and peroxydase isozyme banding patterns in combination with berry colour has led to establish a classification of the 61 autochton varieties into 37 groups including 26 varieties definitely differentiated through the results of this biochemical study.</p>


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