Production of Citrate by Anaerobic Fungi in the Presence of Co-culture Methanogens as Revealed by ¹H NMR Spectrometry

Cheng, Yan Fen,Jin, Wei,Mao, Sheng Yong,Zhu, Wei-Yun Asian Australasian Association of Animal Productio 2013 Animal Bioscience Vol.26 No.10

The metabolomic profile of the anaerobic fungus Piromyces sp. F1, isolated from the rumen of goats, and how this is affected by the presence of naturally associated methanogens, was analyzed by nuclear magnetic resonance spectroscopy. The major metabolites in the fungal monoculture were formate, lactate, ethanol, acetate, succinate, sugars/amino acids and ${\alpha}$-ketoglutarate, whereas the co-cultures of anaerobic fungi and associated methanogens produced citrate. This is the first report of citrate as a major metabolite of anaerobic fungi. Univariate analysis showed that the mean values of formate, lactate, ethanol, citrate, succinate and acetate in co-cultures were significantly higher than those in the fungal monoculture, while the mean values of glucose and ${\alpha}$-ketoglutarate were significantly reduced in co-cultures. Unsupervised principal components analysis revealed separation of metabolite profiles of the fungal mono-culture and co-cultures. In conclusion, the novel finding of citrate as one of the major metabolites of anaerobic fungi associated with methanogens may suggest a new yet to be identified pathway exists in co-culture. Anaerobic fungal metabolism was shifted by associated methanogens, indicating that anaerobic fungi are important providers of substrates for methanogens in the rumen and thus play a key role in ruminal methanogenesis.

Knockdown of Radixin by RNA interference Suppresses the Growth of Human Pancreatic Cancer Cells in Vitro and in Vivo

Chen, Shu-Dong,Song, Mao-Min,Zhong, Zhi-Qiang,Li, Na,Wang, Pi-Lin,Cheng, Shi,Bai, Ri-Xing,Yuan, Hui-Sheng Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.3

Radixin, encoded by a gene on chromosome 11, plays important roles in cell motility, invasion and tumor progression. However, its function in pancreatic cancer remains elusive. In this study, radixin gene expression was suppressed with a lentivirus-mediated short-hairpin RNA (shRNA) method. We found that radixin shRNA caused down-regulation of radixin in PANC-1 cells, associated with inhibition of pancreatic cancer cell proliferation, survival, adhesion and invasive potential in vitro. When radixin-silenced cells were implanted in nude mice, tumor growth and microvessel density were significantly inhibited as compared to blank control cells or nonsense shRNA control cells. Thrombospondin-1 (TSP-1) and E-cadherin were up-regulated in radixin-silenced PANC-1 cells. Our results suggest that radixin might play a critical role in pancreatic cancer progression, possibly through invvolvement of down-regulation of TSP-1 and E-cadherin expression.

Knockdown of Ezrin by RNA Interference Reverses Malignant Behavior of Human Pancreatic Cancer Cells in Vitro

Zhong, Zhi-Qiang,Song, Mao-Min,He, Ying,Cheng, Shi,Yuan, Hui-Sheng Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.8

Background: Pancreatic cancer is one of the most aggressive tumors with a dismal prognosis. The membrane cytoskeletal crosslinker Ezrin participates in several functions including cell proliferation, adhesion, motility and survival. There is increasing evidence that Ezrin is overexpressed in vast majority of malignant tumors and regulates tumor progression. However, its roles in pancreatic cancer remain elusive. Methods: Three pairs of specific Ezrin siRNAs were designed and synthetized and screened to determine the most efficient one for construction of a hairpin RNA plasmid targeting Ezrin. After transfection into the Panc-1 pancreatic cancer cell line, real-time quantitative PCR and Western blotting were performed to examine the expression of mRNA and protein. The MTT method was applied to examine the proliferation and the drug sensibility to Gemcitabine. Flow cytometry was used to assess the cycle and apoptosis, while capacity for invasion was determined with transwell chambers. Furthermore, we detected phosphorylated-Erk1/2 protein and phosphorylated-Akt protein by Western blotting. Results: Real-time quantitative PCR and Western blotting revealed that Ezrin expression was notably down-regulated at both mRNA and protein levels by RNA interference (P< 0.01). Proliferation was inhibited and drug resistance to gemcitabine was improved (P< 0.05). Flow cytometry showed that the proportion of cells in the G1/G0 phase increased (P< 0.01), and in G2/M and S phases decreased (P< 0.05), with no apparent differences in apoptosis (P> 0.05). The capacity for invasion was markedly reduced (P< 0.01). In addition, down-regulating Ezrin expression had no effect on phosphorylated-Akt protein (P>0.05), but could decrease the level of phosphorylated-Erk1/2 protein (P< 0.05). Conclusions: RNA interference of Ezrin could inhibit its expression in the pancreatic cancer cells line Panc-1, leading to a potent suppression of malignant behavior in vitro. Assessment of potential as a target for pancreatic cancer treatment is clearly warranted.

Identification of ssDNA Aptamers Specific for Anti-neuroexcitation Peptide III and Molecular Modeling Studies: Insights into Structural Interactions

Jun Zhu,Jian Wang,Zhen-Cheng Su,Qin Li,Mao-Sheng Cheng,Jing-Hai Zhang 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.9

Twelve ssDNA aptamers specific for a novel recombinant anti-neuroexcitation peptide (ANEPIII) were identified using the SELEX method from a 79-nucleotide ssDNA pool to purify ANEPIII in a more efficient way. To further understand the binding modes between ssDNA and ANEPIII, fully flexible dinucleotides were docked onto the homology-modeled ANEPIII structure. AutoDocking identified favorable binding sites on ANEPIII for nucleotides, which was valuable for designing more potent ligands.

Phenotypic and Molecular Characteristics of Children with Progressive Familial Intrahepatic Cholestasis in South China

Wen Zhang,Ruizhu Lin,Zhikun Lu,Huiying Sheng,Yi Xu,Xiuzhen Li,Jing Cheng,Yanna Cai,Xiaojian Mao,Li Liu 대한소아소화기영양학회 2020 Pediatric gastroenterology, hepatology & nutrition Vol.23 No.6

Purpose: Progressive familial intrahepatic cholestasis (PFIC) is a rare genetic autosomal recessive disease caused by mutations in ATP8B1, ABCB11 or ABCB4. Mutational analysis of these genes is a reliable approach to identify the disorder. Methods: We collected and analyzed relevant data related to clinical diagnosis, biological investigation, and molecular determination in nine children carrying these gene mutations, who were from unrelated families in South China. Results: Of the nine patients (five males, four females) with PFIC, one case of PFIC1, four cases of PFIC2, and four cases of PFIC3 were diagnosed. Except in patient no. 8, jaundice and severe pruritus were the major clinical signs in all forms. γ-glutamyl transpeptidase was low in patients with PFIC1/PFIC2, and remained mildly elevated in patients with PFIC3. We identified 15 different mutations, including nine novel mutations (p.R470HfsX8, p.Q794X and p.I1170T of ABCB11 gene mutations, p.G319R, p.A1047P, p.G1074R, p.T830NfsX11, p.A1047PfsX8 and p.N1048TfsX of ABCB4 gene mutations) and six known mutations (p.G446R and p.F529del of ATP8B1 gene mutations, p.A588V, p.G1004D and p.R1057X of ABCB11 gene mutations, p.P479L of ABCB4 gene mutations). The results showed that compared with other regions, these three types of PFIC genes had different mutational spectrum in China. Conclusion: The study expands the genotypic spectrum of PFIC. We identified nine novel mutations of PFIC and our findings could help in the diagnosis and treatment of this disease.

Identification of ssDNA Aptamers Specific for Anti-neuroexcitation Peptide III and Molecular Modeling Studies: Insights into Structural Interactions

Zhu, Jon,Wang, Jian,Su, Zhen-Cheng,Li, Qin,Cheng, Mao-Sheng,Zhang, Jing-Hai 대한약학회 2008 Archives of Pharmacal Research Vol.31 No.9

Twelve ssDNA aptamers specific for a novel recombinant anti-neuroexcitation peptide (ANEPIII) were identified using the SELEX method from a 79-nucleotide ssDNA pool to purify ANEPIII in a more efficient way. To further understand the binding modes between ssDNA and ANEPIII, fully flexible dinucleotides were docked onto the homology-modeled ANEPIII structure. AutoDocking identified favorable binding sites on ANEPIII for nucleotides, which was valuable for designing more potent ligands.

Alantolactone selectively suppresses STAT3 activation and exhibits potent anticancer activity in MDA-MB-231 cells

Chun, Jaemoo,Li, Rui-Juan,Cheng, Mao-Sheng,Kim, Yeong Shik Elsevier 2015 Cancer letters Vol.357 No.1

<P><B>Abstract</B></P> <P>The important goal of cancer drug discovery is to develop therapeutic agents that are effective, safe, and affordable. In the present study, we demonstrated that alantolactone, which is a sesquiterpene lactone, has potential activity against triple-negative breast cancer MDA-MB-231 cells by suppressing the signal transducer and activator of transcription 3 (STAT3) signaling pathway. Alantolactone effectively suppressed both constitutive and inducible STAT3 activation at tyrosine 705. Alantolactone decreased STAT3 translocation to the nucleus, its DNA-binding, and STAT3 target gene expression. Alantolactone significantly inhibits STAT3 activation with a marginal effect on MAPKs and on NF-κB transcription; however, this effect is not mediated by inhibiting STAT3 upstream kinases. Although SHP-1, SHP-2, and PTEN, which are protein tyrosine phosphatases (PTPs), were not affected by alantolactone, the treatment with a PTP inhibitor reversed the alantolactone-induced suppression of STAT3 activation, indicating that PTP plays an important role in the action of alantolactone. Finally, alantolactone treatment resulted in the inhibition of migration, invasion, adhesion, and colony formation. The <I>in vivo</I> administration of alantolactone inhibited the growth of human breast xenograft tumors. These results provide preclinical evidence to continue the development of alantolactone as a STAT3 inhibitor and as a potential therapeutic agent against breast cancer.</P> <P><B>Highlights</B></P> <P> <UL> <LI> STAT3 is a transcription factor that is a potent regulator of tumorigenesis. </LI> <LI> Alantolactone suppresses constitutive and inducible STAT3 tyrosine phosphorylation. </LI> <LI> Alantolactone inhibits NF-κB translocation to the nucleus. </LI> <LI> Alantolactone inhibits cell migration, invasion, adhesion, and colony formation. </LI> <LI> Alantolactone inhibits the tumor growth of MDA-MB-231 xenografts in mice. </LI> </UL> </P>

Structure-based functional site recognition for p21-activated kinase 4

Jian Wang,Gang Wang,Yu Sha,Dong-Mei Zhao,Feng Li,Mao-Sheng Cheng 대한약학회 2013 Archives of Pharmacal Research Vol.36 No.12

Recently, many molecular modeling methodsare being developed to better understand the principlesunderlying protein folding. In the present study, fullyflexible dinucleotides d(pApA), d(pApC), d(pApG),d(pApT), d(pCpA), d(pCpC), d(pCpG), d(pCpT), d(pGpA),d(pGpC), d(pGpG), d(pGpT), d(pTpA), d(pTpC), d(pTpG)and d(pTpT) were docked onto the surface of p21-activatedkinase 4 (PAK4) kinase domain. The results showedthat automated docking was a useful tool to identify thefunctional sites of PAK4 and it may provide a theoreticalbasis for the interaction data obtained from previousexperiments. Therefore, structure-based docking with fullyflexible dinucleotide probes might be a good tool to predictand annotate the functional sites of enzymes.

The induction of apoptosis by a newly synthesized diosgenyl saponin through the suppression of estrogen receptor-α in MCF-7 human breast cancer cells.

Chun, Jaemoo,Han, Lina,Xu, Mei Ying,Wang, Bo,Cheng, Mao Sheng,Kim, Yeong Shik Pharmaceutical Society of Korea 2014 Archives of Pharmacal Research Vol.37 No.11

<P>Estrogen receptor (ER)-α is an important therapeutic target in the clinical treatment of breast cancer. A potential down-regulator of ER-α, diosgenyl α-L-rhamnopyranosyl-(12)-[β-D-xylopyranosyl-(14)]-α-L-arabinopyranoside is a newly synthesized diosgenyl saponin named compound 22. This study evaluated the in vitro mechanism of compound 22 as an anticancer agent for breast cancer. Our results indicated that compound 22 selectively inhibited proliferation and induced apoptosis in ER-positive MCF-7 cells, compared with ER-negative MDA-MB-231 and MCF-10A cells. Western blot analysis showed that compound 22 decreased the expression of procaspase-3, procaspase-8, and survivin; and increased the expression of Fas ligand and cleaved PARP1 in MCF-7 cells, indicating that compound 22-induced apoptosis was mediated by the extrinsic pathway. This apoptosis was associated with the suppression of ER-α protein and mRNA expression and the inhibition of ER-DNA binding to the estrogen responsive element. Moreover, ER-α mediated gene expression such as c-Myc and cyclin D1 was reduced, and the activation of p38 and ERK 1/2 was significantly decreased after treatment with compound 22 in MCF-7 cells. Taken together, these results demonstrate that compound 22 down-regulates ER-α expression and induces apoptosis through the extrinsic pathway, suggesting that compound 22 may be effective in the treatment of ER-positive breast cancer.</P>

Synthesis of novel diosgenyl saponin analogues and apoptosis-inducing activity on A549 human lung adenocarcinoma

Wang, Bo,Chun, Jaemoo,Liu, Yang,Han, Lina,Wang, Yan-shi,Joo, Eun-Ji,Kim, Yeong-Shik,Cheng, Mao-sheng The Royal Society of Chemistry 2012 Organic & Biomolecular Chemistry Vol.10 No.44

<P>We synthesized a diosgenyl saponin bearing a unique disaccharide from the natural product β-hederin, together with twelve glycosylated derivatives and determined their cytotoxicity against five different human cancer cell lines. Most of them showed weak cytotoxicity, with the exception of compound <B>20</B>, diosgenyl α-<SMALL>L</SMALL>-rhamnopyranosyl-(1→2)-[α-<SMALL>L</SMALL>-arabinopyranosyl-(1→4)]-α-<SMALL>L</SMALL>-arabinopyranoside, which exhibited strong cytotoxicity against A549 cells. The cytotoxicity of <B>20</B> was associated with apoptotic cell death, which was characterized by morphological changes, chromatin condensation, DNA fragmentation, and phosphatidylserine externalization. Compound <B>20</B> induced apoptosis of A549 cells through a caspase-8-mediated extrinsic pathway and a caspase-9-mediated intrinsic pathway. In addition, phosphorylation of JNK increased but the phosphorylation of ERK decreased after treatment with <B>20</B>. These results provide a basic mechanism for the anticancer activity of <B>20</B>.</P> <P>Graphic Abstract</P><P>The diosgenyl trisaccharide greatly induces apoptosis in A549 cells, mainly through an extrinsic pathway. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c2ob26579f'> </P>