Antiproliferative and Antitumorigenic Activity of Toona sinensis Leaf Extracts in Lung Adenocarcinoma

Chih-Jen Yang,Yu-Jung Huang,Cheng-Yuan Wang,Chuan-Sheng Wang,Pei-Hui Wang,Jen-Yu Hung,Tung-Heng Wang,Hseng-Kuang Hsu,Hurng-Wern Huang,S.P. Anand Kumar,Ming-Shyan Huang,Ching-Feng Weng 한국식품영양과학회 2010 Journal of medicinal food Vol.13 No.1

Toona sinensis is a traditional Chinese herb, and the extracts of T. sinensis leaf possess a variety of biological functions. This study attempted to test the antiproliferative effect of TSL-1 (a bioactive fraction of T. sinensis) in H441 cells (lung adenocarcinoma). The data showed that the antiproliferative effect of TSL-1 on H441 cells is prominent using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. TSL-1-induced apoptosis was confirmed by cell morphology, sub-G1 peak accumulation, cleavage of poly(ADP)-ribose polymerase, and propidium iodide-annexin V double staining. Furthermore, decreased Bcl-2 accompanied by increased Bax (in western blotting) was found with TSL-1 treatment of H441 cells. TSL-1 treatment-induced G1 arrest was concurrent with the down-regulation of protein levels of cyclin D1 and cyclin-dependent kinase 4 in H441 cells. Peroral and intraperitoneal administrations of TSL-1 were performed to evaluate the therapeutic efficacy, and peroral administration of TSL-1 was also used to elucidate the therapeutic efficacy in the H441 cell xenograft model in vivo. The data revealed that TSL-1 treatment inhibited H441 tumor growth in both therapeutic and preventive experiments. Taken together, these results demonstrate that TSL-1 possesses the capability of preventing and alleviating lung cancer proliferation in vitro and in vivo with proven nephrological and hepatic safety and has the potential to be developed as an anti–lung cancer drug.

IBM Zigbee Positioning Method for Smart Home Applications

Ming-Hui Jin,Chung-Jung Fu,Chih-Hao Yu,Hung-Ren Lai,Ming-Whei Feng 보안공학연구지원센터 2008 International Journal of Smart Home Vol.2 No.2

This paper presents a smart home system prototype which employs an indoor positioning system called the III Beacon Match (I3BM) positioning method to intelligently trigger the appropriately services for the home members. To overcome the ninja problem in the I3BM method, the signal filtering, adjustment and smooth procedures are proposed. The proposed system prototype employs the Zigbee module to implement the prototypes of the components for the I3BM positioning method, and the prototypes had passed the ZigBee Compliant Platform (ZCP) certification test. The proposed system prototype which intelligently controls the air condition and light system for smart home applications is also verifying in our demo room and in the smart house in National Taiwan University.

Upregulating sirtuin 6 ameliorates glycolysis, EMT and distant metastasis of pancreatic adenocarcinoma with krüppel-like factor 10 deficiency

Tsai Yi-Chih,Chen Su-Liang,Peng Shu-Ling,Tsai Ya-Li,Chang Zuong-Ming,Chang Vincent Hung-Shu,Ch’ang Hui-Ju 생화학분자생물학회 2021 Experimental and molecular medicine Vol.53 No.-

Krüppel-like factor 10 (KLF10) is a tumor suppressor in multiple cancers. In a murine model of spontaneous pancreatic adenocarcinoma (PDAC), additional KLF10 depletion accelerated distant metastasis. However, Klf10 knockout mice, which suffer from metabolic disorders, do not develop malignancy. The mechanisms of KLF10 in PDAC progression deserve further exploration. KLF10-depleted and KLF10-overexpressing PDAC cells were established to measure epithelial-mesenchymal transition (EMT), glycolysis, and migration ability. A murine model was established to evaluate the benefit of genetic or pharmacological manipulation in KLF10-depleted PDAC cells (PDACshKLF10). Correlations of KLF10 deficiency with rapid metastasis, elevated EMT, and glycolysis were demonstrated in resected PDAC tissues, in vitro assays, and murine models. We identified sirtuin 6 (SIRT6) as an essential mediator of KLF10 that modulates EMT and glucose homeostasis. Overexpressing SIRT6 reversed the migratory and glycolytic phenotypes of PDACshKLF10 cells. Linoleic acid, a polyunsaturated essential fatty acid, upregulated SIRT6 and prolonged the survival of mice injected with PDACshKLF10. Modulating HIF1α and NFκB revealed that EMT and glycolysis in PDAC cells were coordinately regulated upstream by KLF10/SIRT6 signaling. Our study demonstrated a novel KLF10/SIRT6 pathway that modulated EMT and glycolysis coordinately via NFκB and HIF1α. Activation of KLF10/SIRT6 signaling ameliorated the distant progression of PDAC. Clinical Trial Registration: ClinicalTrials.gov. identifier: NCT01666184.

Lipopolysaccharide-induced Autophagy Increases SOX2-positive Astrocytes While Decreasing Neuronal Differentiation in the Adult Hippocampus

Liu Wen-Chung,Wu Chih-Wei,Fu Mu-Hui,Tain You-Lin,Liang Chih-Kuang,Chen I-Chun,Hung Chun-Ying,Lee Yu-Chi,Wu Kay L.H. 한국뇌신경과학회 2022 Experimental Neurobiology Vol.31 No.5

Inflammation alters the neural stem cell (NSC) lineage from neuronal to astrogliogenesis. However, the underlying mechanism is elusive. Autophagy contributes to the decline in adult hippocampal neurogenesis under E. coli lipopolysaccharide (LPS) stimulation. SRY-box transcription Factor 2 (SOX2) is critical for NSC self-renewal and proliferation. In this study, we investigated the role of SOX2 in induced autophagy and hippocampal adult neurogenesis under LPS stimulation. LPS (5 ng•100 g-1•hour-1 for 7 days) was intraperitoneally infused into male Sprague–Dawley rats (8 weeks old) to induce mild systemic inflammation. Beclin 1 and autophagy protein 12 (Atg12) were significantly upregulated concurrent with decreased numbers of Ki67- and doublecortin (DCX)-positive cells in the dentate gyrus. Synchronically, the levels of phospho(p)-mTOR, the p-mTOR/mTOR ratio, p-P85s6k, and the p-P85s6k/P85s6k ratio were suppressed. In contrast, SOX2 expression was increased. The fluorescence micrographs indicated that the colocalization of Beclin 1 and SOX2 was increased in the subgranular zone (SGZ) of the dentate gyrus. Moreover, increased S100β-positive astrocytes were colocalized with SOX2 in the SGZ. Intracerebroventricular infusion of 3-methyladenine (an autophagy inhibitor) effectively prevented the increases in Beclin 1, Atg12, and SOX2. The SOX2+-Beclin 1+ and SOX2+-S100β+ cells were reduced. The levels of p-mTOR and p-P85s6k were enhanced. Most importantly, the number of DCX-positive cells was preserved. Altogether, these data suggest that LPS induced autophagy to inactivate the mTOR/P85s6k pathway, resulting in a decline in neural differentiation. SOX2 was upregulated to facilitate the NSC lineage, while the autophagy milieu could switch the SOX2-induced NSC lineage from neurogenesis to astrogliogenesis.

Cognitive Deficits Associated With Dysphagia in Patients With Dementia

( Sun-wung Hsieh ),( Hui-yu Chuang ),( Chih-hsing Hung ),( Chun-hung Chen ) 대한소화기기능성질환·운동학회 2021 Journal of Neurogastroenterology and Motility (JNM Vol.27 No.4

The feasibility of detecting endometrial and ovarian cancer using DNA methylation biomarkers in cervical scrapings

Cheng-Chang Chang,Hui-Chen Wang,Yu-Ping Liao,Yu-Chih Chen,Yu-Chun Weng,Mu-Hsien Yu,Hung-Cheng Lai 대한부인종양학회 2018 Journal of Gynecologic Oncology Vol.29 No.1

Objective: We hypothesized that DNA methylation of development-related genes may occur in endometrial cancer (EC)/ovarian cancer (OC) and may be detected in cervical scrapings. Methods: We tested methylation status by quantitative methylation-specific polymerase chain reaction for 14 genes in DNA pools of endometrial and OC tissues. Tissues of EC/normal endometrium, OC/normal ovary, were verified in training set using cervical scrapings of 10 EC/10 OC patients and 10 controls, and further validated in the testing set using independent cervical scrapings in 30 EC/30 OC patients and 30 controls. We generated cutoff values of methylation index (M-index) from cervical scrapings to distinguish between cancer patients and control. Sensitivity/specificity of DNA methylation biomarkers in detecting EC and OC was calculated. Results: Of 14 genes, 4 (PTGDR, HS3ST2, POU4F3, MAGI2) showed hypermethylation in EC and OC tissues, and were verified in training set. POU4F3 and MAGI2 exhibited hypermethylation in training set were validated in independent cases. The mean M-index of POU4F3 is 78.28 in EC and 20.36 in OC, which are higher than that in controls (6.59; p<0.001 and p=0.100, respectively), and that of MAGI2 is 246.0 in EC and 12.2 in OC, which is significantly higher that than in controls (2.85; p<0.001 and p=0.480, respectively). Sensitivity and specificity of POU4F3/MAGI2 were 83%–90% and 69%–75% for detection of EC, and 61% and 62%–69% for the detection of OC. Conclusion: The findings demonstrate the potential of EC/OC detection through testing for DNA methylation in cervical scrapings.

Hydroxydibenzoylmethane induces apoptosis through repressing ornithine decarboxylase in human promyelocytic leukemia HL-60 cells

Wang, Ming-Fu,Liao, Ya-Fan,Hung, Ying-Cheng,Lin, Chih-Li,Hour, Tzyh-Chyuan,Lue, Ko-Huang,Hung, Hui-Chih,Liu, Guang-Yaw Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.4

Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB), a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (${\Delta}{\psi}_m$), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway.

Genotype­specific methylation of HPV in cervical intraepithelial neoplasia

Yaw-Wen Hsu,Rui-Lan Huang,Po-Hsuan Su,Yu-Chih Chen,Hui-Chen Wang,Chi-Chun Liao,Hung-Cheng Lai 대한부인종양학회 2017 Journal of Gynecologic Oncology Vol.28 No.4

Objective: Hypermethylation of human papillomavirus (HPV) and host genes has beenreported in cervical cancer. However, the degree of methylation of different HPV typesrelative to the severity of the cervical lesions remains controversial. Studies of the degree ofmethylation associated with the host gene and the HPV genome to the severity of cervicallesions are rare. We examined the association of methylation status between host genes andlate gene 1 (L1) regions of HPV16, 18, 52, and 58 in cervical brushings. Methods: Cervical brushings from 147 HPV-infected patients were obtained. The samplescomprised normal (n=28), cervical intraepithelial neoplasia (CIN) 1 (n=45), CIN2 (n=13), andCIN3/carcinoma in situ (n=61). The methylation status of HPV and host genes was measuredusing bisulfite pyrosequencing and quantitative methylation-specific polymerase chainreaction (PCR). Results: The degree of methylation of L1 in HPV16, 18, and 52 was associated with theseverity of the cervical lesion. In HPV52, C-phosphate-G (CpG) sites 6368m, 6405m, and6443m showed significantly higher methylation in lesions ≥CIN3 (p=0.005, 0.003, and0.026, respectively). Methylation of most HPV types except HPV52 (r<−0.1) was positivelycorrelated with the degree of methylation of host genes including PAX1 and SOX1 (0.4≤r≤0.7). Combining HPV methylation with PAX1 methylation improved the clustering for ≥CIN2. Conclusion: Our study showed that the degree of L1 methylation of HPV16, 18, and 52but not 58 is associated with the severity of cervical lesions. The association betweenHPV methylation and host gene methylation suggests different responses of host cellularepigenetic machinery to different HPV genotypes.

Hydroxydibenzoylmethane induces apoptosis through repressing ornithine decarboxylase in human promyelocytic leukemia HL-60 cells

Ming-Fu Wang,Guang-Yaw Liu,Ya-Fan Liao,Ying-Cheng Hung,Chih-Li Lin,Tzyh-Chyuan Hour,Ko-Huang Lue,Hui-Chih Hung 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.4

Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB),a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (Δψ m), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway.

Vimentin Is Involved in Peptidylarginine Deiminase 2-Induced Apoptosis of Activated Jurkat Cells

Hsu, Pei-Chen,Liao, Ya-Fan,Lin, Chin-Li,Lin, Wen-Hao,Liu, Guang-Yaw,Hung, Hui-Chih Korean Society for Molecular and Cellular Biology 2014 Molecules and cells Vol.37 No.5

Peptidylarginine deiminase type 2 (PADI2) deiminates (or citrullinates) arginine residues in protein to citrulline residues in a $Ca^{2+}$-dependent manner, and is found in lymphocytes and macrophages. Vimentin is an intermediate filament protein and a well-known substrate of PADI2. Citrullinated vimentin is found in ionomycin-induced macrophage apoptosis. Citrullinated vimentin is the target of anti-Sa antibodies, which are specific to rheumatoid arthritis, and play a critical role in the pathogenesis of the disease. To investigate the role of PADI2 in apoptosis, we generated a Jurkat cell line that overexpressed the PADI2 transgene from a tetracycline-inducible promoter, and used a combination of 12-O-tetradecanoylphorbol-13-acetate and ionomycin to activate Jurkat cells. We found that PADI2 overexpression reduced the cell viability of activated Jurkat cells in1a dose- and time-dependent manner. The PADI2-overexpressed and -activated Jurkat cells presented typical manifestations of apoptosis, and exhibited greater levels of citrullinated proteins, including citrullinated vimentin. Vimentin overexpression rescued a portion of the cells from apoptosis. In conclusion, PADI2 overexpression induces apoptosis in activated Jurkat cells. Vimentin is involved in PADI2-induced apoptosis. Moreover, PADI2-overexpressed Jurkat cells secreted greater levels of vimentin after activation, and expressed more vimentin on their cell surfaces when undergoing apoptosis. Through artificially highlighting PADI2 and vimentin, we demonstrated that PADI2 and vimentin participate in the apoptotic mechanisms of activated T lymphocytes. The secretion and surface expression of vimentin are possible ways of autoantigen presentation to the immune system.