Combined treatment with verrucarin A and tumor necrosis factor-α sensitizes apoptosis by overexpression of nuclear factor-kappaB-mediated Fas

Jayasooriya, Rajapaksha Gedara Prasad Tharanga,Moon, Dong-Oh,Park, Sang Rul,Choi, Yung Hyun,Asami, Yukihiro,Kim, Mun-Ock,Jang, Jae-Hyuk,Kim, Bo Yeon,Ahn, Jong Seog,Kim, Gi-Young Elsevier 2013 Environmental toxicology and pharmacology Vol.36 No.2

<P><B>Abstract</B></P> <P>Verrucarin A (VA) is a member of the family of macrocyclic trichothecenes, which exhibit anti-cancer and immune-modulating activities. However, VA has not yet been demonstrated to be involved in the sensitization of tumor necrosis factor-alpha (TNF-α)-mediated apoptosis. In the present study, we found that VA triggers TNF-α-induced apoptosis in human breast cancer MDA-MB-231 and MCF-7 cells. In particular, activation of caspas-3 and caspase-8 as well as release of cytochrome <I>c</I> were significantly enhanced in response to the combined treatment with VA and TNF-α (VA/TNF-α) and the pan-caspase inhibitor z-VAD-fmk completely reversed the apoptosis, suggesting that caspases are the main effector molecules in VA/TNF-α-induced apoptosis via the intrinsic and extrinsic pathway. Moreover, we confirmed that enhanced Fas expression plays a critical role, because the Fas-blocking antibody partially inhibited VA/TNF-α-induced apoptosis. VA also increased specific DNA-binding activity of nuclear factor-kappaB (NF-κB) via nuclear translocation of p50 and p65. In addition, pretreatment with the NF-κB inhibitor MG132 blocked VA/TNF-α-induced apoptosis by suppression of NF-κB-dependent Fas expression. These results indicated that VA enhances TNF-α-induced apoptosis via NF-κB-dependent Fas overexpression.</P> <P><B>Highlights</B></P> <P> <UL> <LI> VA sensitizes TNF-α-induced apoptosis in human breast cancer cells. </LI> <LI> VA enhances TNF-α-dependent nuclear translocation of p50 and p65. </LI> <LI> VA/TNF-α triggers NF-κB-dependent Fas-induced cell death. </LI> </UL> </P>

Antagonistic effects of acetylshikonin on LPS-induced NO and PGE2 production in BV2 microglial cells via inhibition of ROS/PI3K/Akt-mediated NF-κB signaling and activation of Nrf2-dependent HO-1

Jayasooriya, R. G.,Lee, K. T.,Choi, Y. H.,Moon, S. K.,Kim, W. J.,Kim, G. Y. Springer Science + Business Media 2015 In vitro cellular & developmental biology Animal Vol.51 No.9

Methanol extract of Hydroclathrus clathratus suppresses matrix metalloproteinase-9 in T24 bladder carcinoma cells by suppressing the NF- κ B and MAPK pathways

Jayasooriya, R.G.P.T.,Choi, Y.H.,Moon, S.-K.,Kim, W.-J.,Kim, G.-Y. Spandidos Publications 2012 ONCOLOGY REPORTS Vol.27 No.2

Camptothecin enhances c-Myc-mediated endoplasmic reticulum stress and leads to autophagy by activating Ca²⁺-mediated AMPK

Jayasooriya, Rajapaksha Gedara Prasad Tharanga,Dilshara, Matharage Gayani,Karunarathne, Wisurumuni Arachchilage Hasitha Maduranga,Molagoda, Ilandarage Menu Neelaka,Choi, Yung Hyun,Kim, Gi-Young Elsevier 2018 Food and chemical toxicology Vol.121 No.-

<P><B>Abstract</B></P> <P>Camptothecin (CPT) from <I>Camptotheca acuminate</I> was discovered for anticancer drugs, which targets topoisomease I. However, whether CPT regulates c-Myc expression has not been understood in endoplasmic reticulum (ER) stress and autophagy. In this study, we found that CPT enhanced c-Myc expression and that the transient knockdown of <I>c-Myc</I> abrogated reactive oxygen species (ROS) generation, which resulted in the accumulation of ER stress-regulating proteins, such as PERK, eIF2α, ATF4, and CHOP. Moreover, the transfection of <I>eIF2α</I>-targeted siRNA attenuated CPT-induced autophagy and decreased the levels of Beclin-1 and Atg7, which indicated that CPT upregulated ER stress-mediated autophagy. In addition, CPT phosphorylated AMPK in response to intracellular Ca<SUP>2+</SUP> release. Ca<SUP>2+</SUP> chelators, ethylene glycol tetraacetic acid and a CaMKII inhibitor, K252a, decreased CPT-induced Beclin-1 and Atg7, and downregulated AMPK phosphorylation, which suggested that CPT-induced Ca<SUP>2+</SUP> release leads to the activation of autophagy through CaMKII-mediated AMPK phosphorylation. CPT also phosphorylated JNK and activated the DNA-binding activity of AP-1; furthermore, knockdown of <I>JNK</I> abolished the expression level of Beclin-1 and Atg7, which implied that the JNK-AP-1 pathway was a potent mediator of CPT-induced autophagy. Our findings indicated that CPT promoted c-Myc-mediated ER stress and ROS generation, which enhances autophagy via the Ca<SUP>2+</SUP>-AMPK and JNK-AP-1 pathways.</P> <P><B>Highlights</B></P> <P> <UL> <LI> CPT induces c-Myc-mediated ROS formation, leading to CHOP expression. </LI> <LI> c-Myc positively regulates CPT-induced ER stress by increasing ROS generation. </LI> <LI> CPT promotes autophagy formation as a result of ER stress. </LI> <LI> CPT promotes autophagy through increased intracellular Ca2+ release. </LI> <LI> CPT induces JNK-mediated autophagy by enhancing AP-1 activity. </LI> </UL> </P>

Molecular chemotherapeutic potential of butein: A concise review

Jayasooriya, Rajapaksha Gedara Prasad Tharanga,Molagoda, Ilandarage Menu Neelaka,Park, Cheol,Jeong, Jin-Woo,Choi, Yung Hyun,Moon, Dong-Oh,Kim, Mun-Ock,Kim, Gi-Young Elsevier 2018 Food and chemical toxicology Vol.112 No.-

<P><B>Abstract</B></P> <P>Butein is a biologically active flavonoid isolated from the bark of <I>Rhus verniciflua</I> Stokes, which is known to have therapeutic potential against various cancers. Notably, butein inhibits cancer cell growth by inducing G<SUB>2</SUB>/M phase arrest and apoptosis. Butein-induced G<SUB>2</SUB>/M phase arrest is associated with increased phosphorylation of ataxia telangiectasia mutated (ATM) and Chk1/2, and consequently, with reduced cdc25C levels. In addition, butein-induced apoptosis is mediated through the activation of caspase-3, which is associated with changes in the expression of Bcl-2 and Bax proteins. Intriguingly, butein sensitizes cells to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis via ERK-mediated Sp1 activation, which promotes the transcription of specific death receptor 5. Butein also inhibits the migration and invasion of human cancer cells by suppressing nuclear factor-κB- and extracellular signal-regulated kinases 1/2-mediated expression of matrix metalloproteinase-9 and vascular endothelial growth factor. Additionally, butein downregulates the expression of human telomerase reverse transcriptase and causes a concomitant decrease in telomerase activity. These findings provide the basis for the pharmaceutical development of butein. The aim of this review is to provide an update on the mechanisms underlying the anticancer activity of butein, with a special focus on its effects on different cellular signaling cascades.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Butein induces apoptosis by activating Bax-caspase-3-PARP axis by PI3K/Akt-NF-κB axis through the inhibition of ROS generation. </LI> <LI> Butein induces G<SUB>2</SUB>/M phase cell cycle arrest by inhibiting ROS-mediated ATM-Chk1/2-Cdc25c-cdc2/cyclin B axis. </LI> <LI> Butein enhances TRAIL-mediated apoptosis by increasing DR5 expression through ERK-mediated Sp1 activation. </LI> <LI> Butein suppresses telomerase activity by inhibiting TERT expression and phosphorylation via c-Myc and PI3K/Akt. </LI> <LI> Butein attenuates angiogenesis, invasion/metastasis, and inflammation by suppressing the NF-κB signal pathway. </LI> </UL> </P>

Fulvic acid promotes extracellular anti-cancer mediators from RAW 264.7 cells, causing to cancer cell death <i>in vitro</i>

Jayasooriya, Rajapaksha Gedara Prasad Tharanga,Dilshara, Matharage Gayani,Kang, Chang-Hee,Lee, Seungheon,Choi, Yung Hyun,Jeong, Yong Kee,Kim, Gi-Young Elsevier 2016 INTERNATIONAL IMMUNOPHARMACOLOGY Vol.36 No.-

<P><B>Abstract</B></P> <P>Fulvic acid (FA) is known to promote electrochemical balance as a donor or a receptor possessing many biomedical functions. Nevertheless, the effect of FA on the anti-cancer activity has not been elucidated. In the current study, we first isolated FA from humus and investigated whether FA regulates immune-stimulating functions, such as production of nitric oxide (NO), in RAW 264.7 cells. Our data showed that FA slightly enhances cell viability in a dose-dependent manner and secretion of NO from RAW 264.7 cells. It upregulated the protein and mRNA expression of inducible NO synthesis (iNOS). In addition, FA enhanced the DNA-binding activity of nuclear factor-κB (NF-κB) in RAW 264.7 cells; the NF-κB inhibitor, pyrrolidine dithiocarbamate (PDTC) effectively attenuated the expression of FA-stimulated iNOS, suggesting that FA stimulates NF-κB to promote iNOS and NO production. Finally, FA-stimulated culture media (FA-CM) from RAW 264.7 cells were collected and MCA-102 fibrosarcoma cells were cultured in this media. The FA-CM augmented MCA-102 fibrosarcoma cell apoptosis; however, an NO inhibitor <I>N</I> <SUP>G</SUP>-monomethyl-<SMALL>L</SMALL>-arginine (NMMA) slightly inhibited the FA-CM-mediated MCA-102 fibrosarcoma cell apoptosis, which was accompanied by low levels of NO. In the present study, we found that FA induces the generation of NO and iNOS in RAW 264.7 cells by inducing NF-κB activation; however, NO did not significantly stimulate MCA-102 fibrosarcoma cell apoptosis in the current study. In addition, FA-CM enhanced cell death in various human cancer cells such as Hep3B, LNCaP, and HL60. Taken together, FA most likely stimulates immune-modulating molecules such as NO and induces cancer cell apoptosis.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Fulvic acid is isolated from humus by acid-base extraction methods. </LI> <LI> Fulvic acid increases proliferation of RAW 264.7 macrophage cells. </LI> <LI> Fulvic acid upregulates the expression of <I>iNOS</I> and NO by inducing NF-κB activity. </LI> <LI> Fulvic acid-stimulated medium of RAW 264.7 macrophages increases apoptosis of MCA-102 fibrosarcoma cells. </LI> </UL> </P>

Implementation of Color Values to Determine the Optimum Maturity Stage for Mechanical Harvesting in ‘Mihong’ Peaches

Sugandhi Hirushika Jayasooriya,Mi Hee Shin,W.M. Upeksha Darshani Wijethun,Gyeong Ho Kim,Ye Ji Moon,Jung Gun Cho,Seul Ki Lee,Si Hyeong Jang,Jin Gook Kim 한국원예학회 2023 한국원예학회 학술발표요지 Vol.2023 No.10

딥러닝 기술을 활용한 복숭아 ‘미황’의 성숙도 자동 분류

Lee Sang Jun,신미희,Jayasooriya L. Sugandhi Hirushika,Wijethunga W.M. Upeksha Darshani,Lee Seul Ki,Cho Jung Gun,Jang Si Hyeong,Cho Byoung-Kwan,김진국 한국원예학회 2024 원예과학기술지 Vol.42 No.1

소비자에게 전달되는 복숭아는 숙도에 따라서 품질이 달라지기 때문에 섭취하기에 적합한 숙도를 고려하여 유통하는 과정이 필요하다. 또한, 숙도는 복숭아의 상품성 및 저장성에 영향을 미칠 수 있어 적합한 수확 시기를 선정하는 작업이 요구되지만, 현재 노지 과수 작목의 숙도 판별에 대한 국내 연구는 미미한 실정이다. 그렇기 때문에 본 연구에서는 딥러닝 객체 탐지 분류모델을 활용하여 복숭아 ‘미황’에 대한 숙도 분류 모델을 개발하였다. 실험실 내부 및 야외에서 촬영된 각 2,800장의 이미지를활용하여 데이터 셋을 구축하였고, 수확 날짜 및 복숭아 과정부(apex)의 색도 a* 값을 기준으로 하는 두 개의 데이터 셋으로구성하여 각 셋의 구분 기준에 따라 미숙, 적숙 그리고 과숙 3개의 class로 분류하였다. Train : Validation : Test 데이터 셋은7 : 2 : 1의 비율로 분류하였고 데이터의 다양성 향상 및 unbalance를 해결하기 위해 augmentation을 실시하였다. 딥러닝 모델은 EfficientNet, YOLOv5 그리고 Vision Transformer를 활용하였으며 EfficientNet에서 가장 우수한 분류 모델 성능을 기록하였다. 날짜 기준 분류 모델은 분류 모델 성능 평가 지표 기준 최저 및 최대 100%의 정확도를 달성하였고, 색도 a* 값 기준 분류모델은 최저 94.7%, 최대 98.2%의 높은 정확도를 보였다. 본 연구에서 개발된 객체 탐지 기반 복숭아 숙도 분류 모델은 향후노지 과수 작목의 숙도 분류를 통한 기계수확 적기 판정 작업에 활용될 수 있을 것으로 판단된다. Peach must be delivered to market when at their proper ripeness, as its fruit quality declines quickly after harvest. Therefore, it is necessary to consider suitable ripeness for consumption and distribution. However, research on ripeness judgments for peaches in the orchard is scarce. This study used deep learning technology to develop a ripeness classification model for ‘Mihwang’ peaches. A dataset was prepared using 2,800 images, each taken from a peach orchard (outside dataset) and a laboratory (inside dataset) with the same fruit. The dataset was constructed based on the harvest date of the peaches and the peach apex’s skin color (a* value). It uses three classes, immature, ripe, and overripe, according to the classification criteria of the two datasets. The model was trained with a ratio of 7:2:1 of training data, validation data, and test data, and image data augmentation was carried out to improve the diversity of the data and to solve any imbalances. Among EfficientNet, YOLOv5, and Vision Transformer, the deep learning model recorded the best classification model performance on EfficientNet. Based on the classification model and performance evaluation index, the harvest-date-based classification model achieved the highest accuracy of 100%. The classification model based on the apex color a* value of peaches showed high accuracy with a minimum rate of 94.7% and a maximum rate of 98.2%. The peach ripeness classification model developed in this study can be used for determining the proper time for the mechanical harvesting of fruit from an orchard.

First report on the gastropod proapoptotic AIF3 counterpart from disk abalone (Haliotis discus discus) deciphering its transcriptional modulation by induced pathogenic stress

Elvitigala, D.A.S.,Jayasooriya, R.G.P.T.,whang, I.,Lee, J. Academic Press 2015 FISH AND SHELLFISH IMMUNOLOGY Vol.47 No.2

Apoptosis inducing factor (AIF) is a flavoprotein that is involved in oxidative phosphorylation and induces apoptosis in eukaryotic cells. There are three isozymes of AIF that have been identified to date, designated as AIF1, AIF2, and AIF3; the human AIF3 is also known as an AIF-like protein (AIFL). This study aimed to identify and characterize a homologue of AIF3 from disk abalone (AbAIF3) that belongs to the phylum Mollusca. The open reading frame (ORF) of AbAIF3 is 1749 base pairs (bp) in length and encodes a protein of 583 amino acids, with a predicted molecular mass of 63.14 kDa. Based on our in-silico analysis, the AbAIF3 protein harbored the typical domain architecture as that of the known AIF family proteins, consisting of N-terminal Rieske and pyridine nucleotide-disulphide oxidoreductase domain. Comparative protein sequence analysis confirmed that AbAIF3 is a homolog of AIF3. Moreover, our phylogenetic analysis revealed that AbAIF3 had a close evolutionary relationship with the molluscan counterparts. Interestingly, AbAIF3 was shown to induce apoptosis in HEK293T cells using transfection assays followed by flow cytometric analysis. In addition, we found that AbAIF3 mRNA expression was ubiquitous in physiologically important tissues, and significantly modulated upon experimental immune stimulations in hemocytes. Collectively, our study illustrates the indispensable role of AbAIF3 in inducing apoptosis in disk abalones, which in turn might be involved in hosts' immune defense mechanisms against microbial infections.

Methanol extract of Codium fragile inhibits tumor necrosis factor-α-induced matrix metalloproteinase-9 and invasiveness of MDA-MB-231 cells by suppressing nuclear factor-κB activation

Dilshara, M.G.,Jayasooriya, R.G.P.T.,Kang, C.H.,Choi, Y.H.,Kim, G.Y. Elsevier 2016 Asian Pacific journal of tropical medicine Vol.9 No.6

<P>Objective: To evaluate whether the methanol extract of Codium fragile (MECF) regulates tumor necrosis factor-alpha (TNF-alpha)-induced invasion of human breast cancer MDA-MB-231 cells by suppressing matrix metalloproteinase-9 (MMP-9). Methods: Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analysis were performed to analyze the expression of MMP-9 and nuclear factor-kappa B (NF-kappa B) subunits, p65 and p50, and I kappa B in MDA-MB-231 cells. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for cell viability. MMP-9 activity and invasion were measured by gelatin zymography and a matrigel invasion assay, respectively. NF-kappa B activity was measured by an electrophoretic mobility shift assay and luciferase activity. Results: MECF had no effect on cell viability up to a concentration of 100 mg/mL in human breast cancer MDA-MB-231 cells regardless of the presence of TNF-alpha. MDA-MB-231 cells that were stimulated with TNF-alpha showed a marked increase of invasion compared to the untreated control, whereas pretreatment with MECF downregulated the TNF-alpha-induced invasion of MDA-MB-231 cells. Additionally, zymography, western blot analysis, and RT-PCR confirmed that MECF decreased TNF-alpha-induced MMP-9 expression and activity which is a key regulator for cancer invasion. According to an electrophoretic morbidity shift assay, pretreatment with MECF in MDA-MB-231 cells significantly decreased the TNF-alpha-induced DNA-binding activity of NF-kappa B, which is an important transcription factor for regulating cancer invasion-related genes such as MMP-9. Furthermore, treatment with MECF sustained the expression of p65 and p50 in response to TNF-alpha in the cytosolic compartment. The luciferase assay demonstrated that MECF attenuated TNF-alpha-induced NF-kappa B luciferase activity. Conclusion: MECF exhibited its anti-invasive capability by downregulating TNF-alpha-induced MMP-9 expression, resulting from the suppression of NF-kappa B activity in the human breast cancer cell line MDA-MB-231.</P>