2021

Permanent URI for this collectionhttps://hdl.handle.net/1807/104116

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Metformin exerts anti-tumor effects via Sonic hedgehog signaling pathway by targeting AMPK in HepG2 cells
(Canadian Science Publishing, 2021-12-17) Hu, Ang; Hu, Zeming; Ye, Jianming; Liu, Yuwen; Lai, Zhonghong; Zhang, Mi; Ji, Weichao; Huang, Lili; Zou, Haohong; Chen, Bin; Zhong, Jianing
Metformin, a traditional first-line pharmacologic treatment for type 2 diabetes, has recently been shown to impart anti-cancer effects on hepatocellular carcinoma (HCC). However, the molecular mechanism of metformin on its antitumor activity is still not completely clear. The Sonic hedgehog (Shh) signaling pathway is closely associated with the initiation and progression of HCC. Therefore, the aim of the current study was to investigate the effects of metformin on the biological behavior of HCC and the underlying functional mechanism of metformin on the Shh pathway. The HCC cellular was induced in HepG2 cells by recombinant human Shh (rhShh). The effects of metformin on proliferation and metastasis were evaluated by proliferation, wound healing and invasion assays in vitro. The mRNA and protein expression levels of proteins related to the Shh pathway were measured by western blotting, quantitative PCR and immunofluorescence staining. Metformin inhibited rhShh-induced proliferation and metastasis. Furthermore, metformin decreased mRNA and protein expression of components of the Shh pathway including Shh, Ptch, Smo and Gli-1. Silencing of AMPK in the presence of metformin revealed that metformin could exert its inhibitory effect via AMPK. Our findings demonstrate that metformin can suppress the migration and invasion of HepG2 cells via AMPK-mediated inhibition of the Shh pathway.
Nisin induces apoptosis in cervical cancer cells via reactive oxygen species generation and mitochondrial membrane potential changes
(Canadian Science Publishing, 2021-12-20) Sadri, Houri; Aghaei, Mahmoud; Akbari, Vajihe
Nisin, an antimicrobial peptide produced by Lactococcus lactis, is widely used as a safe food preservative and has been recently attracting the attention of many researchers as a potential anticancer agent. The cytotoxicity of nisin against HeLa, OVCAR-3, SK-OV-3, and HUVEC cells was evaluated using MTT assay. The apoptotic effect of nisin was identified by Annexin-V/propidium iodide assay, and then it was further confirmed by western blotting analysis, mitochondrial membrane potential (ΔΨm) analysis, and reactive oxygen species (ROS) assay. The MTT assay showed concentration-dependent cytotoxicity of nisin towards cancer cell lines, with the IC50 values of 11.5-23 M, but less toxicity against normal endothelial cells. Furthermore, treatment of cervical cancer cells with 12 M nisin significantly (P
Synergistic Inhibitory Effects of 5-Aza-2'-Deoxycytidine and Cisplatin on Urothelial Carcinoma Growth via Suppressing TGFBI-MAPK Signaling Pathways
(Canadian Science Publishing, 2021-11-24) Shang, Donghao; Li, Gang; Zhang, Caixing; Liu, Yuting
This study is to reveal the gene transcriptional alteration, possible molecular mechanism, and pathways involved in the synergy of 5-aza-2'-deoxycytidine (DAC) and CDDP in UC. Two UC cell lines, 5637 and T24, were used in the study. A cDNA microarray was carried out to identify critical genes in the synergistic mechanism of both agents against UC cells. The results showed that several key regulatory genes, such as interleukin 24(IL24), fibroblast growth factor 1(FGF1), and transforming growth factor beta-induced (TGFBI), were identified and may play critical roles in the synergy of DAC and CDDP in UC. Pathway enrichment suggested that many carcinogenesis-related pathways, such as ECM-receptor interaction and MAPK signaling pathways, may participate in the synergy of both agents. Our results suggested that TGF-β1 stimulates the phosphorylation levels of ERK1/2 and p38 via increasing TGFBI expression, TGFBI-MAPK signaling pathway plays an important role in the synergy of DAC and CDDP against UC. Therefore, we revealed the synergistic mechanism of DAC and CDDP in UC, several key regulatory genes play critical roles in the synergy of combined treatment, and TGFBI-MAPK signaling pathway may be an important potential target of these two agents.
Artesunate inhibits cell proliferation, migration, and invasion of thyroid cancer by regulating the PI3K/AKT/FKHR pathway
(Canadian Science Publishing, 2021-10-28) Xu, Zhiwei; Liu, Xiaojian; Zhuang, Daoping
This study characterized the effects of artesunate on thyroid cancer and partially identified its related molecular mechanism. We determined the effect of artesunate on the proliferation of thyroid cancer cells using the MTT assay, cell colony formation experiments, and western blotting, and used flow cytometry to detect the apoptosis of cancer cells. Using a wound-healing assay, Transwell chamber experiments, and western blotting, we determined the effect of artesunate on cancer cell migration. By co-cultivating artesunate with the PI3K agonist, 740Y-P, we also partially identified the molecular mechanism. Artesunate significantly inhibited the growth, proliferation, migration, and invasion of thyroid cancer cells, and promoted the apoptosis of cancer cells. Using co-cultivation with a PI3K agonist, we found that the inhibitory effect of artesunate on cancer cells was mainly due to suppressing the PI3K/AKT/FKHR signaling pathway. By inhibiting the PI3K/AKT/FKHR signaling pathway, artesunate induced apoptosis of thyroid cancer cells and inhibited their proliferation and migration.
PDGFRβ Modulates Aerobic Glycolysis in Osteosarcoma HOS Cells via the PI3K/AKT/mTOR/c-Myc Pathway
(Canadian Science Publishing, 2021-10-01) Tang, Hu-ying; Guo, Jia-qi; Sang, Bo-tao; Cheng, Jun-ning; Wu, Xiang-mei
Osteosarcoma is a malignant tumor abundant in vascular tissue, and its rich blood supply may have a significant impact on its metabolic characteristics. PDGFRβ is a membrane receptor highly expressed in osteosarcoma cells and vascular wall cells, and its effect on osteosarcoma metabolism needs to be further studied. In this study, we discussed the effect and mechanism of PDGFRβ on the glucose metabolism of osteosarcomaHOS cells. First, GSEA, Pearson's correlation test, and PPI correlation analysis indicated the positive regulation of PDGFRβ on aerobic glycolysis in osteosarcoma. The results of qPCR and western blot further confirmed the prediction of bioinformatics. Glucose metabolism experiments proved that PDGF/PDGFRβ couldeffectively promote the aerobic glycolysis of osteosarcoma cells. In addition, the mitochondrial membrane potential experiment proved that the metabolic change triggered by PDGFRβ was not caused by mitochondrial damage. PI3K pathway inhibitor LY294002 or MEK pathway inhibitor U0126, or Warburg effect inhibitor DCA was used to carry out western blot and glucose metabolism experiments, and the results showed that PDGFBB/PDGFRβ mainly activated the PI3K/AKT/mTOR/ c-Myc pathway to promote aerobic glycolysis in osteosarcoma HOS cells. The newly elucidated role of PDGFRβ provides a novel metabolic therapeutic target for osteosarcoma.
SOAT1 enhances lung cancer invasion through stimulating AKT-mediated mitochondrial fragmentation
(Canadian Science Publishing, 2021-10-14) mo, yijun; lin, lina; Zhang, Jianhua; Yu, Changhui
Sterol O-acyltransferase 1 (SOAT1) is a key enzyme in lipid metabolism, which mediates cholesterol esterification metabolism and is closely associated with many cancers. However, the role of SOAT1 in lung cancer invasion remains unclear. We found that SOAT1 expression was positively correlated with lung cancer invasion. Downregulation of SOAT1 inhibited invasion, mitochondrial fragmentation, AKT phosphorylation, and phospho-Drp (Ser616) in lung cancer cells and promoted intracellular free cholesterol accumulation. Mechanistically, AKT phosphorylation inhibitor MK-2206 alleviated both SOAT1 overexpression or high expression-induced mitochondrial fragmentation and lung cancer cell invasion. Furthermore, intracellular free cholesterol accumulation reduced AKT phosphorylation, SREBP1 mRNA expression, cell invasion, and mitochondrial fragmentation in lung cancer cells with high SOAT1 expression. In summary, our findings suggest that SOAT1 promotes lung cancer invasion activates the PI3K/AKT signaling pathway by downregulating intracellular free cholesterol levels, thereby affecting the regulation of mitochondrial fragmentation.
Cardiovascular protective effects of GLP-1:A focus on the MAPK signaling pathway
(Canadian Science Publishing, 2021-10-13) Zhao, Yu-Yan; Chen, Lin-Hui; Huang, Liang; Li, Yong-Zhen; Yang, Chen; Zhu, Ying; Qu, Shun-Lin; Zhang, Chi
Cardiovascular and related metabolic diseases are significant global health challenges. Glucagon-like peptide 1 (GLP-1) is a brain-gut peptide secreted by ileal endocrine that is now an established drug target in type 2 diabetes (T2DM). GLP-1 targeting agents have been shown not only to treat T2DM, but also to exert cardiovascular protective effects through regulating multiple signaling pathways. The mitogen-activated protein kinase (MAPK) pathway, a common signal transduction pathway for transmitting extracellular signals to downstream effector molecules, is involved in regulating diverse cell physiological processes, including cell proliferation, differentiation, stress, inflammation, functional synchronization, transformation and apoptosis. The purpose of this review is to highlight the relationship between GLP-1 and cardiovascular disease (CVD), and discuss how GLP-1 exerts cardiovascular protective effects through MAPK signaling pathway. This review also discusses the future challenges in fully characterizing and evaluating the CVD protective effects of GLP-1 receptor agonists (GLP-1RA) at the cellular and molecular level. A better understanding of MAPK signaling pathway that are disregulated in CVD may aid in the design and development of promising GLP-1RA.
Emerging regulatory challenges of next-generation synthetic biology
(Canadian Science Publishing, 2021-09-10) Sheahan, Taylor; Wieden, Hans-Joachim
Cell-free synthetic biology is a rapidly developing biotechnology with the potential to solve the world’s biggest problems; however, this promise also has implications for global biosecurity and biosafety.Given the current situation with COVID-19 and its economic impact, capitalizing on the potential of cell-free synthetic biology from an economic, biosafety, and biosecurity perspective contributes to our preparedness for the next pandemic, and urges the development of appropriate policies and regulations, together with the necessary mitigation technologies. Proactive involvement from scientists is necessary to avoid misconceptions and assist in the policy making process.
Characterization of a novel FADS2 transcript variant: Implications for D6D activity regulation in cells
(Canadian Science Publishing, 2021-05-20) Marks, Kristin A.; Fernandes, Maria F.; Diaguarachchige De Silva, Kalsha H.; Tomczewski, Michelle V.; Stark, Ken D.; Duncan, Robin E.
Delta-6-desaturase (D6D) activity is deficient in MCF-7 and other cancer cell lines, but is not explained by FADS2 gene mutations. This deficient activity was not ameliorated by induction of the FADS2 gene, and so we hypothesized that some of the induced FADS2 transcript variants (tv) may play a negative regulatory role. FADS2_tv1 is the reference FADS2 tv, coding for full-length D6D isoform 1 (D6D-iso1), but alternate transcriptional start sites result in FADS2_tv2 and FADS2_tv3 variants encoding D6D-iso2 and D6D-iso3 isoforms, respectively, which lack the catalytically critical N-terminal domain. In MCF-7 cells, FADS2_tv2 and FADS2_tv3 were expressed at significantly higher levels than FADS2_tv1. Overexpression of FADS2_tv2 in HEK293 cells confirmed that D6D-iso2 is non-functional, and co-transfection demonstrated a dominant-negative role for D6D-iso2 in D6D-iso1 activity regulation. FADS2_tv2 was expressed at higher levels than FADS2_tv1 in HeLa, MDA-MB-435, MCF-10A, and HT-29 cells, but at lower levels in A549, MDA-MB-231 and LNCaP cells. Overexpression studies indicated roles for FADS2 variants in proliferation and apoptosis regulation that were also cell-line specific. Increased FADS2_tv2 expression provides a new mechanism to help explain deficient D6D activity in MCF-7 and other cancer cell lines, but it is not a hallmark of malignant cells.
On-cell nuclear magnetic resonance spectroscopy to probe cell surface interactions
(Canadian Science Publishing, 2021-04-25) Phạm, Trần Thanh Tm; Rainey, Jan K
Nuclear magnetic resonance (NMR) spectroscopy allows determination of atomic-level information about intermolecular interactions, molecular structure, and molecular dynamics in the cellular environment. This may be broadly divided into studies focused on obtaining detailed molecular information in the intracellular context (“in-cell”) or those focused on characterizing molecules or events at the cell surface (“on-cell”). In this review, we outline some key NMR techniques applied for on-cell NMR studies through both solution-state and solid-state NMR and survey studies that have used these techniques to uncover key information. We particularly focus on application of on-cell NMR spectroscopy to characterize ligand interactions with cell surface membrane proteins such as G-protein coupled receptors (GPCRs), receptor tyrosine kinases, etc. These techniques allow for quantification of binding affinities, competitive binding assays, delineation of portions of ligands involved in binding, ligand bound-state conformational determination, evaluation of receptor structuring and dynamics, and inference of distance constraints characteristic of the ligand-receptor bound state. Excitingly, it is possible to avoid the barriers of production and purification of membrane proteins while obtaining directly physiologically-relevant information through on-cell NMR. We also provide a briefer survey of the applicability of on-cell NMR approaches to other classes of cell surface molecule.
Chromatin remodeling factor CECR2 forms tissue-specific complexes with CCAR2 and LUZP1
(Canadian Science Publishing, 2021-06-22) Niri, Farshad; Terpstra, Alaina; Lim, Kenji Rowel Quintana; McDermid, Heather
Chromatin remodeling complexes alter chromatin structure to control access to DNA and therefore control cellular processes such as transcription, DNA replication, and DNA repair. CECR2 is a chromatin remodeling factor that plays an important role in neural tube closure and reproduction. Loss-of-function mutations in Cecr2 result primarily in the perinatal lethal neural tube defect exencephaly, with non-penetrant mice that survive to adulthood exhibiting subfertility. CECR2 forms a complex with ISWI proteins SMARCA5 and/or SMARCA1, but further information on the structure and function of the complex is not known. We therefore have identified candidate components of the CECR2-containing remodeling factor (CERF) complex in embryonic stem (ES) cells through mass spectroscopy. Both SMARCA5 and SMARCA1 were confirmed to be present in CERF complexes in ES cells and testis. However, novel proteins CCAR2 and LUZP1 are CERF components in ES cells but not testis. This tissue specificity in mice suggests these complexes may also have functional differences. Furthermore, LUZP1, loss of which is also associated with exencephaly, appears to play a role in stabilizing the CERF complex in ES cells. Keywords: CECR2, LUZP1, CCAR2, Chromatin remodeling factor, Neural tube defects
Aberrant sphingomyelin 31P-NMR signatures in giant cell tumour of bone
(Canadian Science Publishing, 2021-06-01) Quiroz-Acosta, Tayde; Flores-Martinez, Yazmin Montserrat; Becerra-Martnez, Elvia; Prez-Hernndez, Elizabeth; Prez-Hernndez, Nury; Bauelos-Hernndez, Angel Ernesto
An understanding of the biochemistry of the giant cell tumour of bone (GCTB) provides an opportunity for the development of prognostic markers and identification of therapeutic targets. Based on metabolomic analysis, we proposed glycerophospholipid metabolism as the altered pathway in GCTB and the objective of this study was to identify these altered metabolites. Using phosphorus-31 nuclear magnetic resonance spectroscopy (
GCN5 enables HSP12 induction promoting chromatin remodeling not histone acetylation.
(Canadian Science Publishing, 2021-06-07) Antonazzi, Francesca; Di Felice, Francesca; Camilloni, Giorgio
Regulation of stress responsive genes represents one of the best examples of gene induction and the relevance and involvement of different regulators may change for a given gene depending on the challenging stimulus. HSP12 gene is induced by very different stimuli, however the molecular response to the stress has been characterized in detail only for heat shock treatments. In this work we want to verify whether, the regulation of transcription induced by oxidative stress, utilizes the same epigenetic solutions relative to those employed in heat shock response. We also monitored HSP12 induction employing spermidine, a known acetyltransferase inhibitor, and observed an oxidative stress that synergizes with spermidine treatment. Our data show that during transcriptional response to H2O2, histone acetylation and chromatin remodeling occur. However, when the relevance of Gcn5p on these processes was studied, we observed that induction of transcription is GCN5 dependent and this does not rely on histone acetylation by Gcn5p despite its HAT activity. Chromatin remodeling accompanying gene activation is rather GCN5 dependent. Thus, GCN5 controls HSP12 transcription after H2O2 treatment by allowing chromatin remodeling and it is only partially involved in HSP12 histone acetylation regardless its HAT activity.
Isovitexin attenuates tumor growth in human colon cancer cells through the modulation of apoptosis and epithelial-mesenchymal transition via PI3K/Akt/mTOR signaling pathway
(Canadian Science Publishing, 2021-06-11) Zhu, Hao; Zhao, Na; Jiang, Maozhu
Isovitexin, a biologically active flavone C-glycosylated derivative, has a variety of biological activities. We aimed to identify the effect of isovitexin (Isov) on colon cancer. The human colonic epithelial cell (HCEC), and cancer cells were treated with Isov, Cell Counting Kit-8 (CCK8) was used to detect cell proliferation and calculate half-inhibitory concentration (IC50). The biological activity of cancer cells were assessed. The tumor size and volume were recorded. The expression levels of proteins were analyzed by western blot. Isov inhibited cancer cells proliferation, while had little cytotoxicity on HCEC. Isov significantly attenuated cell proliferation, migration, invasion and epithelial-mesenchymal transition (EMT), induced cell apoptosis, and that trends were blocked by insulin-like growth factor-1 (IGF-1) treated. The expression levels of phosphorylated phosphatidylinositol 3-kinasep (p-PI3K), phosphorylated protein kinase B (p-Akt), phosphorylated mammalian target of rapamycin (p-mTOR), and B-cell lymphoma-2 (Bcl-2) evidently decreased when treated with Isov, while the levels of Bcl2-associated X (Bax), and caspase-3 significantly increased. After Isov treatment, the tumor volume and weight were decreased, the levels of p-PI3K, p-Akt, p-mTOR, and Bcl-2 significantly decreased in tumor tissues. Our finding demonstrated that Isov could inhibit cancer cells migration, invasion and EMT. Isov maybe a new potentially treatment medicine for colon cancer.
Saikosaponin-d protects against liver fibrosis by regulating Estrogen receptor-β/NLRP3 inflammasome pathway
(Canadian Science Publishing, 2021-04-09) Lin, Liubing; Zhou, Mengen; Que, Renye; Chen, Yirong; Liu, Xiaolin; Zhang, Kehui; Shi, Zhe; Li, Yong
Liver fibrosis is the ultimate common pathway in most types of chronic liver damage characterized by imbalanceof extracellular matrix degradation and synthesis. Saikosaponin-d (SSd) possesses anti-inflammatory and anti-liver fibrosiseffects. However, the underlying mechanism of SSd in repressing hepaticstellatecells(HSCs) activation remains unclear. Here we found that SSdalleviated remarkably carbon tetrachloride (CCl4)-induced liver fibrosis, as evidenced by decreased collagen level and profibrotic markers (COl1a1 and α-smooth muscle actin (SMA)) expression. SSdrepressed CCl4-induced NOD-like receptor family pyrin-domain-containng-3 (NLRP3) activation in fibrotic livers, as suggested by decreased level of NLRP3, IL-18, and IL-β. The primary HSCs of CCl4 mice exhibited a significant increase in profibrotic markers expression and NLRP3 activation, but SSd treatment reversed the effect. SSd also repressed TGF-β-induced profibrotic markers expression and NLRP3 activation in vitro. Mechanistically, TGF-β decreased the expression of Estrogen receptor-β (ERβ) in HSCs, whereas SSd treatment reversed the effect. ERβ inhibition enhanced NLRP3 activation in HSCs. More important, ERβ or NLRP3 inhibition destroyed partially the function of SSd on anti-liver fibrosis. In summary, the current data suggest that SSd prevents hepatic fibrosis through regulating ERβ/NLRP3 inflammasome pathway, and suggest SSd as a potential agent for treating liver fibrosis.
S-Allylcysteine Inhibits Chondrocyte Inflammation to Reduce Human Osteoarthritis via Targeting RAGE, TLR4, JNK and Nrf2 Signaling: Comparison with Colchicine
(Canadian Science Publishing, 2021-04-25) Elmazoglu, Zubeyir; Aydın Bek, Zehra; Saribas, Sanem Gulistan; zoğul, Candan; Goker, Berna; Bitik, Berivan; Aktekin, Cem Nuri; Karasu, imen
Discovery of new pharmacological agents is needed to control the progression of osteoarthritis (OA) characterized by progressive joint cartilage damage. Human OA chondrocyte cultures (OAC) were either applied to S-Allyl cysteine (SAC), a sulfur-containing amino acid derivative, or colchicine, an ancient anti-inflammatory therapeutic, for 24 hours. SAC or colchicine did not change viability at 1 nM-10 M but inhibited p-JNK/pan-JNK. While SAC seems to be more effective, both agents inhibited reactive oxygen species (ROS), 3-nitrotyrosine (3-NT), lipid-hydroperoxides (LPO), advanced lipoxidation end-products (ALEs as 4-hydroxy-2-nonenal, HNE) and advanced glycation end-products (AGEs), and increased glutathione-peroxidase (GPx) and type-II-collagen (COL2). IL-1β, IL-6 and osteopontin (OPN) were more strongly inhibited by SAC than in colchicine. In contrast, TNF-α was inhibited only by SAC, and COX2 only by colchicine. Casp-1/ICE, GM-CSF, receptor for advanced glycation end-products (RAGE) and toll-like receptors (TLR4) were inhibited by both agents, but bone morphogenetic protein 7 (BMP7) was partially inhibited by SAC while induced by colchicine. The nuclear factor erythroid 2-related factor 2 (Nrf2) was induced by SAC; in contrast it was inhibited by colchicine. Although exerting opposite effects on TNF-α, COX2, BMP7 and Nrf2, SAC and colchicine exhibit anti-osteoarthritic properties in OAC by modulating redox sensitive inflammatory signaling.
Substitution of histone H3 arginine 72 to alanine leads to deregulation of isoleucine biosynthesis in budding yeast Saccharomyces cerevisiae
(Canadian Science Publishing, 2021-04-09) Thakre, Pilendra Kumar; Sahu, Rakesh Kumar; Tomar, Raghuvir Singh
Histone residues play an essential role in the regulation of various biological processes. In the present study, we have utilized the H3/H4 histone mutant library to probe functional aspects of histone residues in amino acid biosynthesis. We found that histone residue H3R72 plays a crucial role in the regulation of isoleucine biosynthesis. Substitution of arginine residue (H3R72) of histone H3 to alanine (H3R72A) renders yeast cells unable to grow in the minimal media. Histone mutant H3R72A requires the external supplementation of either isoleucine, serine, or threonine for the growth in minimal media. We also observed that H3R72 residue and leucine amino acid in synthetic complete media might play a crucial role in determining the intake of isoleucine and threonine in yeast. Further, gene deletion analysis of ILV1 and CHA1 in H3R72A mutant confirmed that isoleucine is the sole requirement for growth in minimal medium. Altogether, we have identified that histone H3R72 residue may be crucial for yeast growth in the minimal medium by regulating isoleucine biosynthesis through the Ilv1 enzyme in budding yeast Saccharomyces cerevisiae.
Predicting human RNA quadruplex helicases through comparative sequence approaches and helicase mRNA interactome analyses
(Canadian Science Publishing, 2021-02-07) Dupas, Steven J.; Gussakovsky, Daniel; Wai, Alvan; Brown, Mira J.F.; Hausner, Georg; McKenna, Sean A.
RNA quadruplexes are non-canonical nucleic acid structures involved in several human disease states and are regulated by a specific subset of RNA helicases. Given the difficulty in identifying RNA quadruplex helicases due to the multifunctionality of these enzymes, we sought to provide a comprehensive in silico analysis of features found in validated RNA quadruplex helicases to predict novel human RNA quadruplex helicases. Using the 64 human RNA helicases, we correlated their amino acid compositions with subsets of RNA quadruplex helicases categorized by varying level of evidence of RNA quadruplex interaction. Utilizing phylogenetic and synonymous/non-synonymous substitution analyses, we identified an evolutionarily conserved pattern involving predicted intrinsic disorder and a previously identified motif. We analyzed available next generation sequencing data to determine which RNA helicases were directly interacting with predicted RNA quadruplex regions intracellularly and elucidated a relationship with miRNA binding sites adjacent to RNA quadruplexes. Finally, we employed a phylogenetic analysis of all 64 human RNA helicases to establish how RNA quadruplex detection and unwinding activity may be conserved among helicase subfamilies. This work furthers understanding of commonalities between RNA quadruplex helicases and provides support for the future validation of several human RNA helicases.
1 E-Cadherin knockdown induces cancer stem cell-like phenotype and drug resistance
(Canadian Science Publishing, 2021-02-05) Sharma, Anuka; Kaur, Harmandeep; De, Renaissa; Srinivasan, Radhika; Pal, Arnab; Bhattacharyya, Shalmoli
Cervical cancer is one of the leading causes of mortality amongst women in developing countries and therapy resistance is the main reason for its treatment failure. Recent advances suggest that cancer stem cells (CSCs) are critically involved in regulating the chemo resistant behavior of cervical cancer cells. In our study the CSC phenotype cells were isolated and the expression of stem cell marker and epithelial-mesenchymal transition (EMT) associated gene was confirmed by various assays. However, these CSC phenotype cells cannot be cultured for further cytotoxicity studies. So, we tried to establish a CSC model in cervical cancer cells. We performed the siRNA-mediated knockdown of E-cadherin (E-cad) in these cells and studied EMT associated stem cell-like properties in them. We also performed dose dependent cell viability assay using clinically relevant drugs such as cisplatin, cyclopamine and GANT58 to analyze the drug resistant behavior of these cancer cells. We found that E-cad knockdown induces EMT in cervical cancer cells imparting stem-cell like characteristics along with enhanced tumorsphere formation, migration, invasion ability and drug resistance. This is the first study to establish a CSC model in cervical cancer cells by knockdown of E-cad which can be utilized for development of anti-cancer therapies.
Cetrimonium bromide promotes lipid clearance via TFEB-mediated autophagy-lysosome activation in hepatic cells
(Canadian Science Publishing, 2021-01-18) Liu, Zhenxing; Wang, Xu; Shi, Zhichen; Xu, Junting; Lin, Jieru; Li, Dianlong; Zhang, Xinpeng; Li, Yuyin; Zhao, Qing; Tao, Li; Diao, Aipo
Autophagy plays a key role in the metabolism of macromolecules by the lysosomal degradative machinery. The transcription factor EB (TFEB) regulates autophagosome biogenesis and lysosome function, and promoting TFEB activity has emerged as a potential strategy for the treatment of metabolic disorders. Here, we describe that cetrimonium bromide (CTAB), a quaternary ammonium compound, promotes autophagy and lysosomal biogenesis by inducing the nuclear translocation of TFEB in hepatic cells. shRNA-mediated TFEB knockdown inhibits CTAB-induced autophagy and lysosomal biogenesis. Mechanistically, CTAB treatment inhibits the Akt-mTORC1 signaling pathway. Moreover, CTAB treatment markedly promotes lipid metabolism in both palmitate and oleate-treated HepG2 cells, and this promotion was attenuated by the depletion of TFEB. Altogether, our results indicate that CTAB activates the autophagy-lysosome pathway by inducing the nuclear translocation of TFEB via the inhibition of mTORC1 signaling. These results deepen our understanding of TFEB function and provide new insights into CTAB-mediated lipid metabolism.

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