作者：
Gonzalez, Kelly D.;Hill, Kathleen A.;Li, Kai;Li, Wenyan;Scaringe, William A.;...
期刊：
Human Mutation,2007年28(1):69-80 ISSN：1059-7794
通讯作者：
Sommer, Steve S.
作者机构：
City Hope Natl Med Ctr, Beckman Res Inst, Dept Mol Genet, Duarte, CA 91010 USA.;Univ Western Ontario, Dept Biol, London, ON, Canada.;Nanhua Univ, SNP Inst, Hengyang, Hunan, Peoples R China.;City Hope Natl Med Ctr, Bioinformat Grp, Dept Mol Genet, Duarte, CA 91010 USA.;[Sommer, Steve S.] City Hope Natl Med Ctr, Beckman Res Inst, Dept Mol Genet, 1500 E Duarte Rd, Duarte, CA 91010 USA.
通讯机构：
[Sommer, Steve S.] C;City Hope Natl Med Ctr, Beckman Res Inst, Dept Mol Genet, 1500 E Duarte Rd, Duarte, CA 91010 USA.
摘要：
Microindels, defined as mutations that result in a colocalized microinsertion and microdeletion with a net gain or loss of between 1 and 50 nucleotides, may be an important contributor to cancer. We report the first comprehensive analysis of somatic microindels. Our large database of mutations in the lacI transgene of Big Blue® mice contains 0.5% microindels, 2.8% pure microinsertions, and 11.5% pure microdeletions. There appears to be no age, gender, or tissue-type specificity in the frequency of microindels. Of the independent somatic mutations that result in a net in-frame insertion or deletion, microindels are responsible for 13% of protein expansions and 6% of protein contractions. These in-frame microindels may play a crucial role in oncogenesis and evolution via “protein tinkering” (i.e., modest expansion or contraction of proteins). Four characteristics suggest that microindels are caused by unique mechanisms, not just simple combinations of the same mechanisms that cause pure microinsertions and pure microdeletions. First, microinsertions and microdeletions commonly occur at hotspots, but none of the 30 microindels are recurrent. Second, the sizes of the deletions and insertions in microindels are larger and more varied than in pure microdeletions and pure microinsertions. Third, microinsertions overwhelmingly repeat the adjacent base (97%) while the insertions in microindels do so only infrequently (17%). Fourth, analysis of the sequence contexts of microindels is consistent with unique mechanisms including recruitment of translesion DNA synthesis polymerases. The mouse somatic microindels have characteristics similar to those of human germline microindels, consistent with similar causative mechanisms in mouse and human, and in soma and germline. Hum Mutat 28(1), 69–80, 2007. Published 2006 Wiley-Liss, Inc.
摘要：
To develop a minipig model of type 2 diabetes that simulates the common manifestations of the metabolic abnormalities and resembles the kidney pathology of type 2 diabetes in the human population, male Chinese Bama minipigs were divided into 2 groups (5 in each) and fed with a control diet (CD) or high-fat/ high-sucrose/ high-cholesterol diet (HFSCD) for 5 months. The biochemical parameters of blood and urine, and the oral glucose tolerance test were monitored after the feeding program. The insulin resistance was estimated by the HOMA-IR index and the glucose elimination constant (K(G)), and beta-cell function by the HOMA-beta index and the acute insulin response (AIR). Glomerulosclerosis index (GSI) was semi-quantitated by the degree of glomerular lesions in kidney sections stained with Masson trichrome. Extracellular matrix deposition in the kidney was examined by the protein expression of type IV collagen, connective tissue growth factor (CTGF) and matrix metalloproteinases 2 (MMP-2) using immunohistochemistry. Feeding HFSCD to minipigs markedly caused hyperglycaemia, hyperinsulinaemia and dyslipidaemia. HOMA-IR was significantly increased while HOMA-beta, AIR and K(G) were obviously decreased in the HFSCD group compared with control group. Microalbuminuria, glucosuria and moderate glomerulosclerosis were exhibited in HFSCD-fed minipigs. The expression of type IV collagen and CTGF was elevated whereas that of MMP-2 was reduced in the kidneys of HFSCD group compared with the CD group. We concluded that feeding HFSCD to Chinese Bama minipigs for 5 months can induce humanoid type 2 diabetes and early-stage diabetic nephropathy, and accelerate extracellular matrix deposition and glomerulosclerosis.
通讯机构：
[Lei, Xiaoyong] U;Univ S China, Inst Pharm & Pharmacol, Hengyang 421001, Peoples R China.
关键词：
small interfering RNA;adenocarcinoma;Bcl-2;A549/DDP;apoptosis
摘要：
Bcl-2 is overexpressed in a variety of human tumors and is involved in tumorigenesis and chemoresistance. In this study, we investigated the inhibitory effect of the hairpin Bcl-2 small interfering (si)RNA on the expression of the Bcl-2 gene in the cisplatin (DDP)-resistant human lung adenocarcinoma cell line A549/DDP, and the effect of Bcl-2 siRNA on drug sensitization in A549/DDP cells. Bcl-2 siRNA and negative siRNA plasmids were constructed and stably transfected into A549/DDP cells. Reverse transcription-polymerase chain reaction, immunofluorescence microscopy and Western blot analysis were used to detect the target gene expression. Spontaneous cell apoptosis was detected by acridine orange and ethidium bromide staining. Drug sensitivity of the cells to DDP and diallyl disulfide (DADS) was analyzed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry. Expression levels of Bcl-2 mRNA and protein in siRNA stable transfectants were clearly reduced compared with negative siRNA transfectants and untreated cells. MTT results indicated that Bcl-2 transfectants had a higher cell inhibition rate after treatment with 0.2-200 μg/ml DDP or 50-200 μM DADS. Flow cytometry revealed increased apoptosis in Bcl-2 siRNA cells. After the addition of 20 μg/ml DDP or 100 μM DADS, siRNA targeting of the Bcl-2 gene specifically down-regulated gene expression in A549/DDP cells, increased spontaneous apoptosis, and sensitized cells to DDP and DADS.
作者：
Gu, Dongqing;Scaringe, William A.;Li, Kai;Saldivar, Juan-Sebastian;Hill, Kathleen A.;...
期刊：
Human Mutation,2007年28(8):760-770 ISSN：1059-7794
通讯作者：
Sommer, Steve S.
作者机构：
City Hope Natl Med Ctr, Clin Mol Diagnost Lab, Dept Mol Diagnosis, Duarte, CA 91010 USA.;Nanhua Univ, SNP Inst, Hunan, Peoples R China.;Univ Western Ontario, Dept Biol, London, ON, Canada.;[Sommer, Steve S.] City Hope Natl Med Ctr, Beckman Res Inst, Dept Mol Genet, 1500 E Duarte Rd, Duarte, CA 91010 USA.
通讯机构：
[Sommer, Steve S.] C;City Hope Natl Med Ctr, Beckman Res Inst, Dept Mol Genet, 1500 E Duarte Rd, Duarte, CA 91010 USA.
作者机构：
So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, Springfield, IL 62702 USA.;Nanhua Univ, Sch Life Sci & Technol, Div Pharmacoproteom, Inst Pharm & Pharmacol, Hengyang 421001, Hunan, Peoples R China.;[Cao, Deliang] So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, 913 N Rutledge St, Springfield, IL 62702 USA.
通讯机构：
[Cao, Deliang] S;So Illinois Univ, Sch Med, Dept Med Microbiol Immunol & Cell Biol, SimmonsCooper Canc Inst, 913 N Rutledge St, Springfield, IL 62702 USA.
关键词：
Acrolein;Aldo-keto reductase family 1 B10;Aldose reductase-like-1;Clonogenic growth;Lactate dehydrogenase
摘要：
Acrolein is a highly reactive α,β-unsaturated aldehyde produced endogenously during lipid peroxidation and naturally distributed pervasively in living environments, posing serious threats to human health if not properly metabolized. In this study, we report aldose reductase–like-1 (ARL-1) as a novel enzyme that catalyzes the reduction of acrolein and protects cells from their toxicity. Using purified ARL-1 protein, we determined its enzymatic activity in response to acrolein and defined its steady-state kinetics with Km and Vmax at 0.110 ± 0.012mM and 3122.0 ± 64.7 nmol/mg protein/min, respectively. By introducing a functional Enhanced Green Fluorescent Protein (EGFP)/ARL-1 fusion protein into 293T cells, we demonstrated that plating efficiency in liquid culture and focus formation in soft agar increased by more than 60% (p < 0.05), compared to the vector control cells. More significantly, at a low dose of 5μM acrolein, EGFP/ARL-1 expression enhanced both plating efficiency and focus formation by more than threefold, and the foci (in soft agar) of 293T cells expressing EGFP/ARL-1 were significantly larger than those of the vector control cells. At high concentrations of acrolein (25 and 50μM), EGFP/ARL-1 protein prevented oncotic death of 293T cells induced by acrolein. In summary, our data demonstrated for the first time that the ARL-1 protein protects 293T cells from acrolein toxicity. Due to the high toxicity and wide distribution of acrolein, this finding is important to the understanding of its detoxification mechanisms.
摘要：
To investigate the effects of recombinant human adiponectin on the metabolism of diabetic swine induced by feeding a high-fat/high-sucrose diet (HFSD), diabetic animal models were constructed by feeding swine with HFSD for 6 months. The effects of recombinant adiponectin were assessed by detecting the change of plasma glucose levels by commercially available enzymatic method test kits and evaluating the insulin sensitivity by oral glucose tolerance test (OGTT). About 1.5 g purified recombinant adiponectin was produced using a 15-liter fermenter. A single injection of purified recombinant human adiponectin to diabetic swine led to a 2- to 3-fold elevation in circulating adiponectin, which triggered a transient decrease in basal glucose level (P<0.05). This effect on glucose was not associated with an increase in insulin level. Moreover, after adiponectin injection, swine also showed improved insulin sensitivity compared with the control (P<0.05). Adiponectin might have the potential to be a glucose-lowering agent for metabolic disease. Adiponectin as a potent insulin enhancer linking adipose tissue and glucose metabolism could be useful to treat insulin resistance.
期刊：
International Journal of Cardiology,2007年120(3):331-337 ISSN：0167-5273
通讯作者：
Fu, Mingde
作者机构：
[Fu, Mingde] Sichuan Univ, W China Hosp, Lab Endocrinol & Metab, Chengdu 610041, Peoples R China.;Nanhua Univ, Dept Biochem & Mol Biol, Hengyang, Hunan, Peoples R China.;Hoist Grp Postdoctoral Work Stn, Chengdu, Sichuan, Peoples R China.
通讯机构：
[Fu, Mingde] S;Sichuan Univ, W China Hosp, Lab Endocrinol & Metab, Chengdu 610041, Peoples R China.
关键词：
2-Dimensional gel electrophoresis-immunodetection;Combined hyperlipidemia;HDL subclasses;Hypercholesterolemia
摘要：
Background: Alterations in plasma lipid levels can influence the composition, content, and distribution of plasma lipoprotein subclasses that effect atherosclerosis risk. Hypercholesterolemia and combined hyperlipidemia are common forms of atherogenic dyslipoproteinemia. This study evaluates the alterations of high-density lipoprotein (HDL) subclasses in hypercholesterolemic and combined hyperlipidemic subjects. Methods: Apolipoprotein A-I contents of plasma HDL subclasses were quantitated by 2-dimensional gel electrophoresis in 242 normolipidemic subjects, 66 hypercholesterolemic subjects and 59 combined hyperlipidemic subjects. Results: Compared with the normolipidemic subjects, apolipoprotein A-I contents of small-sized pre-beta(1)-HDL, HDL3c, HDL(3b)and HDL(3a)were significantly higher in both hypercholesterolemic subjects (p <.01, p <.05, p <.01 and p <.05, respectively) and combined hyperlipidemic subjects (p <.0l,p <.05,p <.01 and p <.01, respectively). In contrast, apolipoprotein A-I contents of large-sized HDL2a and HDL2b were significantly lower in hypercholesterolemic subjects (p <.05 and p <.01, respectively) as well as combined hyperlipidemic subjects (p <.01 and p <.01, respectively). In addition, pre-beta(1)-HDL increased significantly (p <.05) while HDL2a and HDL2b decreased significantly (p <.05 and p <.0 1, respectively) in combined hyperlipidemic group versus hypercholesterolemic subjects. With the elevation of triglyceride levels, pre-beta(1)-HDL, and HDL3a. increased successively, however, HDL2a and HDL2b decreased successively in subjects with total cholesterol levels greater than 240 mg/dl. Conclusions: The particle size of HDL shifted towards smaller size in hypercholesterolemic subjects, and that the shift was more prominent in combined hyperlipidemic subjects. The alternations mentioned above indicate that HDL maturation might be abnormal, and reverse cholesterol transport (RCT) might be weakened. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
摘要：
Bcl-XL is overexpressed in a variety of human tumors and is involved in tumorigenesis and chemoresistance. This study investigated the inhibitory effect of the hairpin Bcl-XL small interfering RNA (siRNA) on the expression of the Bcl-XL gene in the cisplatin (DDP)-resistant human lung adenocarcinoma cell line A549/DDP, and the effect of Bcl-XL siRNA on drug sensitization in A549/DDP cells. Bcl-XL siRNA and negative siRNA plasmids were constructed and stably transfected into A549/DDP cells. Reverse transcription-polymerase chain reaction and Western blot analysis were used to detect the target gene expression. Spontaneous apoptosis of cells was detected by acridine orange and ethidium bromide staining. Drug sensitivity of the cells to DDP was analyzed with dimethylthiazol-diphenyltetrazolium bromide (MTT) and flow cytometry. Expression levels of Bcl-XL mRNA and protein in siRNA stable transfectants were clearly reduced as compared with negative siRNA transfectants and untreated cells. MTT results indicated that Bcl-XL transfectants had a higher cell inhibition rate than the negative vector or untreated cells after treatment with 0.2–200 μg/ml DDP. Flow cytometry revealed increased apoptosis in Bcl-XL siRNA cells. After the addition of 20 μg/ml DDP, siRNA targeting of the Bcl-XL gene specifically down-regulated gene expression in A549/DDP cells, increased spontaneous apoptosis, and sensitized cells to DDP. The results showed that Bcl-XL siRNA contributed to an increase of DDP-induced cell death in non-small-cell lung cancer and sensitized cells to DDP, leading to increased the effectiveness of the drug in treating non-small-cell lung cancer.
期刊：
International Journal of Cancer,2007年121(10):2301-2306 ISSN：0020-7136
通讯作者：
Cao, Deliang
作者机构：
So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, Springfield, IL 62794 USA.;Nanhua Univ, Sch Life Sci & Technol, Inst Pharm & Pharmacol, Div Pharmacoproteom, Hengyang, Hunan, Peoples R China.;Mem Hosp Carbondale, Dept Pathol, Carbondale, IL USA.;[Cao, Deliang] So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.
通讯机构：
[Cao, Deliang] S;So Illinois Univ, Sch Med, SimmonsCooper Canc Inst, Dept Med Microbiol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.
关键词：
aldose reductase-like-1;aldo-keto reductase family 1 B 10;reactive carbonyls;gene silencing;clonogenic growth
摘要：
Aldo-keto reductase family 1 B10 (AKR1B10), a member of aldo-keto reductase superfamily, is overexpressed in human hepatocellular carcinoma, lung squamous cell carcinoma and lung adenocarcinoma. Our previous study had demonstrated that the ectopic expression of AKR1B10 in 293T cells promotes cell proliferation. To evaluate its potential as a target for cancer intervention, in the current study we knocked down AKR1B10 expression in HCT-8 cells derived from a colorectal carcinoma, using chemically synthesized small interfering RNA (siRNA). The siRNA 1, targeted to encoding region, downregulated AKR1B10 expression by more than 60%, and siRNA 2, targeted to 3' untranslational region, reduced AKR1B10 expression by more than 95%. AKR1B10 silencing resulted in approximately a 50% decrease in cell growth rate and nearly 40% suppression of DNA synthesis. More importantly, AKR1B10 downregulation significantly reduced focus formation rate and colony size in semisolid culture, indicating the critical role of AKR1B10 in HCT-8 cell proliferation. Recombinant AKR1B10 protein showed strong enzymatic activity to acrolein and crotonaldehyde, with K(m) = 110.1 +/- 12.2 microM and V(max) = 3,122.0 +/- 64.7 nmol/mg protein/min for acrolein and K(m) = 86.7 +/- 14.3 microM and V(max) = 2,647.5 +/- 132.2 nmol/mg protein/min for crotonaldehyde. AKR1B10 downregulation enhanced the susceptibility of HCT-8 cells to acrolein (25 microM) and crotonaldehyde (50 microM), resulting in rapid oncotic cell death characterized with lactate dehydrogenase efflux and annexin-V staining. These results suggest that AKR1B10 may regulate cell proliferation and cellular response to additional carbonyl stress, thus being a potential target for cancer intervention.
期刊：
Journal of Diabetes Research,2007年2007:67435 ISSN：2314-6745
通讯作者：
Lin, GP
作者机构：
[Lin, Guo-Ping; Jiang, Tao; Hu, Xiao-Bo; Qiao, Xin-Hui; Tuo, Qin-Hui] Univ S China, Sch Life Sci & Technol, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Lin, GP ] ;Univ S China, Sch Life Sci & Technol, Hengyang 421001, Hunan, Peoples R China.
摘要：
The Siraitia grosvenorii polysaccharide (SGP) from the Siraitia grosvenorii (Swingle) was isolated and purified. The therapeutic effects of SGP on diabetic rabbits induced by feeding high fat/high sucrose chow were studied. After administration of SGP for 4 weeks, the fasting blood glucose (FBG), plasma insulin levels (INS), plasma total cholesterol (TC), triglyceride (TG), and HDL-C were assayed. The results showed that administration of SGP can significantly decrease plasma total cholesterol, triglyceride, and glucose levels; and increase HDL-C levels after 4 weeks of treatment. The antihyperglycaemic effect of SGP at dose of 100 mg.kg(-1) bw was the most significant in three dosage groups. Furthermore, SGP could restore the blood lipid levels of diabetic rabbits (P<.05). These data indicate that SGP not only ameliorates the lipid disorder, but also lowers plasma glucose levels. So SGP have obvious glucose-lowering effect on hyperglycaemic rabbits induced by feeding high fat/high sucrose chow, its mechanism may be related to amelioration of lipid metabolism and restoring the blood lipid levels of hyperglycaemic rabbits.
