scholarly journals cAMP stimulates SLC26A3 activity in human colon by a CFTR-dependent mechanism that does not require CFTR activity

2018 ◽  
Author(s):  
Chung-Ming Tse ◽  
Jianyi Yin ◽  
Varsha Singh ◽  
Rafiquel Sarker ◽  
Ruxian Lin ◽  
...  
Keyword(s):  
Cell Model ◽  
Cancer Cell Line ◽  
Human Colon ◽  
Colon Cancer Cell Line ◽  
Proximal Colon ◽  
Transport Assay ◽  
Normal Human ◽  
Regulatory Effects ◽  
Camp Stimulation ◽  
Dependent Mechanism

AbstractBackground & AimsDRA (SLC26A3) is an electroneutral Cl-/HCO3- exchanger that is present in the apical domain of multiple intestinal segments. An area that has continued to be poorly understood is related to DRA regulation in acute cAMP-related diarrheas, in which DRA appears to be both inhibited as part of NaCl absorption and stimulated to contribute to increased HCO3- secretion. Different cell models expressing DRA have shown that cAMP inhibits, stimulates or does not affect its activity.MethodsThis study reevaluated cAMP regulation of DRA using new “tools” including a successful knockout cell model, a specific DRA inhibitor (DRAinh-A250), specific antibodies, and a transport assay that did not rely on non-specific inhibitors. The studies compared DRA regulation in colonoids made from normal human colon with regulation in the colon cancer cell line, Caco-2.ResultsDRA is an apical protein in human proximal colon, differentiated colonoid monolayers and Caco-2 cells. It is glycosylated and appears as two bands. cAMp(forskolin) acutely stimulated DRA activity in human colonoids and Caco-2 cells. In these cells, DRA is the predominant apical Cl-/HCO3- exchanger and is inhibited by DRAinh-A250 with IC50 of 0.5 μmol/L and 0.2 µmol/L, respectively. However, there was no effect of cAMP in HEK293/DRA cells that lacked CFTR. When CFTR was expressed in HEK293/DRA cells, cAMP also stimulated DRA activity. In all cases, cAMP stimulation of DRA was not inhibited by CFTRinh-172.ConclusionsDRA is acutely stimulated by cAMP by a process that is CFTR-dependent but appears to be one of multiple regulatory effects of CFTR that does not require CFTR activity.

Rotundic Acid Regulates the Effects of Let-7f-5p on Caco2 Cell Proliferation

2020 ◽  
Vol 20 ◽  
Author(s):  
Yuan Feng ◽  
Xinran Liu ◽  
Yueqing Han ◽  
Mantian Chen ◽  
Lin Zhang ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Colony Formation ◽  
Cancer Cell Line ◽  
Human Colon ◽  
Colon Cancer Cell Line ◽  
Inhibitory Effect ◽  
Human Colon Cancer ◽  
Caco2 Cell ◽  
Study Results ◽  
Rotundic Acid

Background & Objective: Nowadays, the interaction between natural products and microRNAs provides a promising field for exploring the chemo preventive agents for various cancers.As a member of microRNAs, the expression of let-7f-5p is universally down regulated in colorectal cancer (CRC). The present study aimed to uncover the function of let-7f-5p in the proliferation of human colon cancer cell line Caco2 and explored chemo preventive agents from natural resources that can prevent the development of CRC. Methods: Herein, Caco2 cells were transfected with let-7f-5p mimic and inhibitor to manipulate let-7f-5p levels, and the expression of let-7f-5p wasper formed by RT‑qPCR. Next, we determined how let-7f-5p regulates Caco2 cell proliferation by using MTT, wound-healing, cell cycle,and colony formation assays.Besides, to further understand the effect of let-7f-5p, we evaluated the protein level of AMER3 and SLC9A9 by using western blotting assays. Results: The results showed a suppressive function of let-7f-5p on Caco2 cell proliferation and then put forward a triterpenoid (rotundic acid, RA) which significant antagonized the effect of cell proliferation, restitution after wounding,and colony formation caused by let-7f-5p. Moreover, the western blot results further indicated that the inhibitory effect of RA might be due to its suppressive role in let-7f-5p-targeted AMER3 and SLC9A9 regulation. Conclusion: Our validation study results confirmed that let-7f-5p was a potent tumor suppressor gene of Caco2 cell proliferation,and RA showed as a regulator of the effect oflet-7f-5p on cell proliferation and then could be a potential chemo preventive agent for CRC treatment.

scholarly journals Biogenic synthesis of gold nanoparticles using Argemone mexicana L. and their cytotoxic and genotoxic effects on human colon cancer cell line (HCT-15)

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Kailas D. Datkhile ◽  
Satish R. Patil ◽  
Pratik P. Durgawale ◽  
Madhavi N. Patil ◽  
Dilip D. Hinge ◽  
...  
Keyword(s):  
Cell Line ◽  
Caspase 3 ◽  
Cancer Cell Line ◽  
Human Colon ◽  
Colon Cancer Cell Line ◽  
Colon Cancer Cell ◽  
Human Colon Cancer Cell ◽  
Genotoxic Effects ◽  
Argemone Mexicana ◽  
Human Colon Cancer

Abstract Background Nanomedicine has evolved as precision medicine in novel therapeutic approach of cancer management. The present study investigated the efficacy of biogenic gold nanoparticles synthesized using Argemone mexicana L. aqueous extract (AM-AuNPs) against the human colon cancer cell line, HCT-15. Results Biosynthesis of AM-AuNPs was determined by ultraviolet-visible spectroscopy and further characterized by transmission electron microscopy, X-ray diffraction, and Fourier transition infrared spectroscopy analysis. The cytotoxic activity of AM-AuNPs was assessed by the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, whereas genotoxicity was evaluated by the DNA fragmentation assay. The expression of apoptosis regulatory genes such as p53 and caspase-3 was explored through semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) and western blotting to evidence apoptotic cell death in HCT-15 cells. Biogenic AM-AuNPs inhibited cell proliferation in HCT-15 cell line with a half maximal inhibitory concentration (IC50) of 20.53 μg/mL at 24 h and 12.03 μg/mL at 48 h of exposure. The altered cell morphology and increased apoptosis due to AM-AuNPs were also evidenced through nuclear DNA fragmentation and upregulated expression of p53 and caspase-3 in HCT-15 cells. Conclusion The AM-AuNPs may exert antiproliferative and genotoxic effects on HCT-15 cells by cell growth suppression and induction of apoptosis mediated by activation of p53 and caspase-3 genes.

scholarly journals Antitumor activity, cell cycle arrest and apoptosis induction in human colon cancer cell line by Primula macrophylla extracts

2017 ◽  
Vol 12 (2) ◽  
pp. 4
Author(s):  
Di-Wen Shou ◽  
Yue-Lin Zheng
Keyword(s):  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Methanol Extract ◽  
Cancer Cell Line ◽  
Human Colon ◽  
Colon Cancer Cell Line ◽  
Human Colon Cancer Cell ◽  
Apoptosis Induction ◽  
Human Colon Cancer ◽  
Cycle Arrest

<p class="Abstract">The primary objective of the current work was to investigate the antitumor potential of <em>Primula macrophylla</em> extracts in human colon cancer cell line (Colo-205) along with evaluating the effects on apoptosis induction, cell cycle arrest and mitochondrial membrane potential. Cell viability was assessed by tetrazolium-based MTT assay. Flow cytometry measurement was carried out to assess the effect of the extract on cell cycle phase distribution and mitochondrial transmembrane potential. Results showed that <em>P.  macrophylla</em> methanol extract was effective and exhibited highest cell growth inhibition (IC<sub>50</sub> value, 26.17 μg/mL). Methanol extract significantly increased the side-scattering profile of Colo-205 cells in concentration-dependent pattern. Exposure of Colo-205 cells with different concentrations of the methanol extract (0-80 μg/mL) caused dose-dependent G0/G1 cell cycle arrest along with inducing apoptotic cascade by increasing the population of cells at G0/G1 phase. Furthermore, methanol extract treatment caused an increase in mitochondrial membrane depolarization in Colo-205 cells.</p><p class="Abstract"><strong>Video Clip of Methodlogy</strong> (3 min 17 sec): <a href="https://youtube.com/v/yy-rCjDN830">Click</a>  <a href="https://youtube.com/watch?v=yy-rCjDN830">If failed</a></p>

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