scholarly journals Human small cell lung cancer cell line (NCI-H146) tumor behaviour on the chicken embryo chorioallantoic membrane after the sodium valproate treatment

2017 ◽  
Vol 26 (2) ◽  
pp. 144
Author(s):  
Lina Šlekienė ◽  
Raminta Mozūraitė ◽  
Ruta Vosyliūtė ◽  
Ingrida Balnytė ◽  
Angelija Valančiūtė

Small cell lung cancer (SCLC) is an aggressive form of cancer with 5-year survival rate and poor prognosis. Since patients' initial response to therapy is rapidly followed by a relapse with the drug-resistant disease, new therapies are required. The aim was to evaluate whether H146 cell line tumor was able to maintain the morphological pattern when grafted on the chicken embryo chorioallantoic membrane (CAM). Also, to evaluate the morphological changes in the CAM after grafting the tumor without or with treatment with sodium valproate (NaVP). The cell culture of the commercial NCIH146 cell line was used for the formation of the tumor. The tumor of 1x106 cells and I type rat tail collagen was dropped onto the absordable sponge and instantly grafted on CAM. After 5 days of incubation chicken embryos were sacrificed and CAMs were cut out. Standard H&E staining and immunohistochemistry (CD56) were performed. The histomorphometrical analysis of non-treated (n=5), 4 mM NaVP-treated (n=5) and 8 mM (n=5) of NaVP-treated groups was performed. The thickness of the CAM was measured in all the investigated groups in the areas under the onplant and in the neighbouring sites. H146 cells are able to maintain morphological appearance on the CAM and retained the expression of specific profile for CD56. H146 cell tumor increased the thickness of the CAM under the tumor in the non-treated group (509.2±184μm). After treatment with 4 mM and 8 mM of NaVP CAM thickness decreased (362±232.6 μm and 147.8±98.4 μm respectively; p<0.0001). The effect of NaVP on CAM increased with increasing solution concentrations of 4 mM and 8 mM. CAM thickness in the non-treated group neighbouring to the tumor site was 139.1±138.4 μm, in 4 mM NaVP-treated group – 104.9±81 μm and in 8 mM NaVP-treated group CAM thickness reached only 70.8±51.5 μm. CAM thickness in the non-treated group significantly differs compared to 4 mM and 8 mM of NaVP – the treated groups (p<0.05, p<0.0001). Difference between both NaVP-treated groups is also significant (p<0.05). CAM is a suitable model for in vivo analysis of human H146 cell line formed tumor. Cells grafted on the CAM preserved their morphology and expressed characteristic immunohistochemical markers. CAM mesenchyme proliferation was suppressed under the influence of NaVP in concentration–dependent manner.

Dose-Response ◽  
2018 ◽  
Vol 16 (2) ◽  
pp. 155932581877248 ◽  
Author(s):  
Lina Šlekienė ◽  
Donatas Stakišaitis ◽  
Ingrida Balnytė ◽  
Angelija Valančiūtė

The study aims to test the effect of different sodium valproate (NaVP) doses on small cell lung cancer NCI-H146 cells tumor in chicken embryo chorioallantoic membrane (CAM) model. Xenografts were investigated in the following groups: nontreated control and 5 groups treated with different NaVP doses (2, 3, 4, 6, and 8 mmol/L). Invasion of tumors into CAM in the nontreated group reached 76%. Tumors treated with 8 mmol/L NaVP doses significantly differed in tumor invasion frequency from the control and those treated with 2 mmol/L ( P < .01). The calculated probability of 50% tumor noninvasion into CAM was when tumors were treated with 4 mmol/L of NaVP. Number of p53-positive cells in tumors was significantly reduced when treated with NaVP doses from 3 to 8 mmol/L as compared with control; number of EZH2-positive cells in control significantly differed from all NaVP-treated groups. No differences in p53- and EZH2-positive cell numbers were found among 4, 6, and 8 mmol/L NaVP-treated groups. Invaded tumors had an increased N-cadherin and reduced E-cadherin expression. The results indicate the increasing NaVP dose to be able to inhibit tumors progression. Expression of p53 and EZH2 may be promising target markers of therapeutic efficacy evaluation.


2017 ◽  
Vol 7 (3) ◽  
Author(s):  
Chukwumaobim D.U. Nwokwu ◽  
Sameera R. Samarakoon ◽  
Desiree N. Karunaratne ◽  
Nuwanthi P. Katuvawila ◽  
Meran K. Ediriweera ◽  
...  

2021 ◽  
Vol 27 ◽  
pp. 101089
Author(s):  
Kazuo Ohara ◽  
Shintaro Kinoshita ◽  
Jun Ando ◽  
Yoko Azusawa ◽  
Midori Ishii ◽  
...  

Biomolecules ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 917
Author(s):  
Valeria Sorrenti ◽  
Agata Grazia D’Amico ◽  
Ignazio Barbagallo ◽  
Valeria Consoli ◽  
Salvo Grosso ◽  
...  

In order to maintain redox homeostasis, non-small-cell lung cancer (NSCLC) increases the activation of many antioxidant systems, including the heme-oxygenase (HO) system. The overexpression of HO-1 has been often associated with chemoresistance and tumor aggressiveness. Our results clearly showed an overexpression of the HO-1 protein in A549 NSCLC cell lines compared to that in non-cancerous cells. Thus, we hypothesized that “off-label” use of tin mesoporphyrin, a well-known HO activity inhibitor clinically used for neonatal hyperbilirubinemia, has potential use as an anti-cancer agent. The pharmacological inhibition of HO activity caused a reduction in cell proliferation and migration of A549. SnMP treatment caused an increase in oxidative stress, as demonstrated by the upregulation of reactive oxygen species (ROS) and the depletion of glutathione (GSH) content. To support these data, Western blot analysis was performed to analyze glucose-6-phosphate dehydrogenase (G6PD), TP53-induced glycolysis and the apoptosis regulator (TIGAR), and the glutamate cysteine ligase catalytic (GCLC) subunit, as they represent the main regulators of the pentose phosphate pathway (PPP) and glutathione synthesis, respectively. NCI-H292, a subtype of the NSCLC cell line, did not respond to SnMP treatment, possibly due to low basal levels of HO-1, suggesting a cellular-dependent antitumorigenic effect. Altogether, our results suggest HO activity inhibition may represent a potential target for selective chemotherapy in lung cancer subtypes.


2004 ◽  
Vol 95 (2) ◽  
pp. 142-148 ◽  
Author(s):  
Tatsuya Yoshimasu ◽  
Teruhisa Sakurai ◽  
Shoji Oura ◽  
Issei Hirai ◽  
Hirokazu Tanino ◽  
...  

2018 ◽  
Vol 51 (6) ◽  
pp. 2938-2954 ◽  
Author(s):  
Jing Shen ◽  
Shoubo Cao ◽  
Xin Sun ◽  
Bo Pan ◽  
Jingyan Cao ◽  
...  

Background/Aims: Sonodynamic therapy (SDT) is expected to be a new method to solve the clinical problems caused by advanced metastasis in patients with lung cancer. The use of ultrasound has the advantage of being noninvasive, with deep-penetration properties. This study explored the anti-tumor effect of SDT with a new sonosensitizer, sinoporphyrin sodium (DVDMS), on the human small cell lung cancer H446 cell line in vitro and in vivo. Methods: Absorption of DVDMS was detected by a fluorescence spectrophotometer, and DVDMS toxicity was determined using a Cell Counting Kit-8. Mitochondrial membrane potential (MMP) was assessed using the JC-1 fluorescent probe. Cell apoptosis was measured by flow cytometry, and apoptosis-related proteins were detected by western blotting. The expression of cytokines was measured using an enzyme-linked immunosorbent assay and quantitative real-time PCR. To verify the in vitro results, we detected tumor volumes and weight changes in a xenograft nude mouse model after DVDMS-SDT. Hematoxylin and eosin staining was used to observe changes to the tumor, heart, liver, spleen, lung, and kidney of the mice, and immunohistochemistry was used to examine changes in the expression of tumor CD34 and receptor-interacting protein kinase-3 (RIP3), while terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was used to observe apoptosis in tumor tissues. Results: DVDMS-SDT-treated H446 cells increased the rate of cellular apoptosis and the levels of reactive oxygen species (ROS), cleaved caspase-3, cleaved caspase-8, cleaved caspase-9, and caspase-10, and decreased the levels of MMP, RIP3, B-cell lymphoma 2, vascular endothelial growth factor, and tumor necrosis factor-α. The sonotoxic effect was mediated by ROS and was reduced by a ROS scavenger (N-acetyl-L-cysteine). In the in vivo mouse xenograft model, DVDMS-SDT showed efficient anti-cancer effects with no visible side effects. Conclusion: DVDMS-SDT induced apoptosis in H446 cells, in part by targeting mitochondria through the mitochondria-mediated apoptosis signaling pathway, and the extrinsic apoptosis pathway was also shown to be involved. Both apoptosis and changes in RIP3 expression were closely related to the generation of ROS. DVDMS-SDT will be advantageous for the management of small cell lung cancer due to its noninvasive characteristics.


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