Inhibition of lncRNA KCNQ1OT1 Improves Apoptosis and Chemotherapy Drug Response in Small Cell Lung Cancer by TGF-β1 Mediated Epithelial-to-Mesenchymal Transition

Deyu Li,Qin Tong,Yuane Lian,Zhizhong Chen,Yaru Zhu,Weimei Huang,Yang Wen,Qiongyao Wang,Shumei Liang,Man Li,Jianjing Zheng,Zhenhua Liu,Huanxin Liu,Linlang Guo 대한암학회 2021 Cancer Research and Treatment Vol.53 No.4

Purpose Drug resistance is one of the main causes of chemotherapy failure in patients with small cell lung cancer (SCLC), and extensive biological studies into chemotherapy drug resistance are required. Materials and Methods In this study, we performed lncRNA microarray, in vitro functional assays, in vivo models and cDNA microarray to evaluate the impact of lncRNA in SCLC chemoresistance. Results The results showed that KCNQ1OT1 expression was upregulated in SCLC tissues and was a poor prognostic factor for patients with SCLC. Knockdown of KCNQ1OT1 inhibited cell proliferation, migration, chemoresistance and promoted apoptosis of SCLC cells. Mechanistic investigation showed that KCNQ1OT1 can activate transforming growth factor-β1 mediated epithelial-to-mesenchymal transition in SCLC cells. Conclusion Taken together, our study revealed the role of KCNQ1OT1 in the progression and chemoresistance of SCLC, and suggested KCNQ1OT1 as a potential diagnostic and prognostic biomarker in SCLC clinical management.

Removal of Feedback Inhibition of Corynebacterium glutamicum Phosphoenolpyruvate Carboxylase by Addition of a Short Terminal Peptide

Deyu Xu,Jing Zhao,Guoqiang Cao,Jinyu Wang,Qinggang Li,Ping Zheng,Shuxin Zhao,Jibin Sun 한국생물공학회 2018 Biotechnology and Bioprocess Engineering Vol.23 No.1

Phosphoenolpyruvate carboxylase (PEPC) catalyzes the carboxylation of phosphoenolpyruvate (PEP) in the presence of bicarbonate to form oxaloacetate (OAA), and it plays an important role in high-efficient production of OAA-derived metabolites such as lysine, glutamate and succinate. However, PEPCs often suffered from serious feedback inhibition by various metabolic effectors like aspartate. Here, the feedback inhibition of PEPC from Corynebacterium glutamicum was removed by adding a short terminal peptide like His-tag. The effect of His-tag location on the structure and important properties such as activity, thermostability and feedback inhibition of PEPC has been investigated. The purified untagged PEPC, Nterminal His-tagged PEPC (PEPC-N-His) and C-terminal His-tagged PEPC (PEPC-C-His) were characterized. PEPCN- His (439.71/sec/mM) showed a 1.26 and 186-fold higher catalytic efficiency than untagged PEPC (348.59/sec/mM) and PEPC-C-His (2.36/sec/mM), respectively. Both PEPCN- His and untagged PEPC were significantly inhibited by aspartate at the concentrations above 4 mM (residual activities < 10%), while PEPC-C-His was almost desensitized to aspartate within 10 mM (around 90% of residual activity). Structural analysis showed that the extension of C-terminus may cause steric hindrance for aspartate binding with enzymes, leading to the deregulation of feedback inhibition of PEPC-C-His. This study provides a deeper understanding of the effect of terminal fragments on the structure and function of PEPCs, and helps to engineer the feedback inhibition of PEPCs and structurally similar enzymes.

Effects of exploration and molecular mechanism of CsV on eNOS and vascular endothelial functions

Zuo, Deyu,Jiang, Heng,Yi, Shixiong,Fu, Yang,Xie, Lei,Peng, Qifeng,Liu, Pei,Zhou, Jie,Li, Xunjia Techno-Press 2022 Advances in nano research Vol.12 No.5

This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

Probability Fuzzy Attribute Implications for Interval-Valued Fuzzy Sets

Yanhui Zhai,Deyu Li,Kaishe Qu 보안공학연구지원센터 2012 International Journal of Database Theory and Appli Vol.5 No.4

Fisetin Inhibits Cell Proliferation and Induces Apoptosis via JAK/STAT3 Signaling Pathways in Human Thyroid TPC 1 Cancer Cells

Ying Liang,Deyu Kong,Yi Zhang,Siqi Li,Yan Li,Anuradha Ramamoorthy,Junfeng Ma 한국생물공학회 2020 Biotechnology and Bioprocess Engineering Vol.25 No.2

Thyroid cancer is the most important malignant tumor reported in human populations where, its treatment remains undeveloped. Fisetin, a plant flavonoid exhibits several pharmacological properties like antioxidant, antiinflammatory, and anticancer function. In the existing study, we assessed fisetin mediated cytotoxic effects and action potential of fisetin on cell proliferation in TPC-1 human cancer cells. Also, examined the apoptosis in TPC-1 cells by reactive oxygen species (ROS) generation, the mitochondrial membrane potential (MMP) and apoptotic morphological changes through AO/EtBr staining. Additionally, we analyzed the effects of fisetin through ELISA analysis to evaluate the caspases expression and studied JAK 1 and STAT3 signaling molecule in TPC1 cells. Our results demonstrated that fisetin stimulated apoptosis, which confirmed through reduced cell viability, improved ROS generation, altered MMP and cell cycle phases in TPC-1 cells. Further, the fisetin up-regulated the expression of caspase (3, 8, and 9) expressions in TPC-1 cells. Also, we observed the fisetin down-regulated the JAK 1 and STAT3 expression in TPC1 cells. Thus, the fisetin induced apoptosis in TPC-1 cells by the induction of oxidative damage and enhanced caspases expression by down-regulating JAK 1 and STAT3 signaling molecules. Hence, the fisetin would be considered as a beneficial therapeutic agent for the thyroid cancer treatment.

Identification and differential expression of two isogenes encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase in Glycine max

Man Zhang,Deyue Yu,Kai Li,Jianyu Liu 한국식물생명공학회 2012 Plant biotechnology reports Vol.6 No.4

Plant 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) were considered to be encoded by single copy. In this study, we successfully isolated two DXR isogenes, designated GmDXR1 and GmDXR2, from soybean. The multicopy nature of DXRs in soybean was supported by Southern blot. Alignment of the two GmDXRs showed the presence of the N-terminal transit peptide for plastids, the conserved cleavage site, prolinerich region, and the NADPH-binding motif. Phylogenetic analysis revealed GmDXR1 and GmDXR2 belonged to different branches of angiosperm DXRs clusters. Expression pattern analysis indicated that GmDXR1 was expressed in all analyzed organs except the pod walls, with the highest level in seeds, whereas no message was detected for GmDXR2. In response to heat stress, GmDXR1 showed declined transcript levels in the experiments; in contrast,GmDXR2 was highly responsive, with mRNA accumulation peaking at 6 h after treatment. We also demonstrated that both GmDXRs were localized in chloroplasts. Overexpression of GmDXRs induced increased contents of various isoprenoids (chlorophyll, carotenoids, and gibberellins),but with reduced level of ABA, indicating that GmDXRs participate in the control of differential isoprenoid biosynthesis of the MEP pathway. This work provides a new insight into the wide presence of multicopies of DXR enzyme in plants.

Effect of Different Potassium Level on Growth and Fruit Yield of Camellia Oleifera Chang-Lin Series

Lu You,Zhi Li,Dekui Niu,Dongnan Hu,Wenyuan Zhang,Xiaomin Guo,Deyue Meng 한국토양비료학회 2014 한국토양비료학회 학술발표회 초록집 Vol.2014 No.6

A rapid and quantitative fluorescent microsphere immunochromatographic strip test for detection of antibodies to porcine reproductive and respiratory syndrome virus

Yanqiu Wei,Baozhi Yang,Yunlong Li,Yongcheng Duan,Deyu Tian,Baoxiang He,Chuangfu Chen,Wenjun Liu,Limin Yang 대한수의학회 2020 Journal of Veterinary Science Vol.21 No.4

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.