摘要：
Lung cancer is the leading cause of cancer-related mortality in humans worldwide. Moreover, the overall 5-year survival rate is only 15%. Pathologically almost 80% of all lung cancer cases are non-small cell lung cancer (NSCLC). Cancer-associated fibroblasts (CAFs) have been found to exist in a large number of NSCLCs. CAFs have been proven to promote tumor progression, metastasis and resistance to therapy through paracrine effects in most solid tumors. In the present study, firstly we isolated CAFs from patient tissues and demonstrated that they promoted cell proliferation and chemoresistance to cisplatin in the lung cancer cell lines A549 and 95D in a paracrine manner. Secondly, using ELISA and quantative PCR, we found that a higher amount of stromal cell derived factor 1 (SDF-1) existed in the CAFs rather than that observed in the normal fibroblasts (NFs). Thirdly, we detected that SDF-1 facilitated lung cancer cell proliferation and drug resistance via the CXCR4-mediated signaling pathway which involved NF-kappa B and Bcl-xL. Moreover, we also confirmed that the expression level of SDF-1 in the CAFs was negatively regulated by microRNA mir-1 through microRNA overexpression and quantitative PCR. Overall, our data provide one explanation for the effects of CAFs on lung cancer cells. Meanwhile, our results also suggest CAFs as a potential therapeutic target in tumor treatment.
作者：
Bu, Yiwen;Cai, Guoshuai;Shen, Yi;Huang, Chenfei;Zeng, Xi;...
期刊：
Cancer Letters,2016年383(2):261-271 ISSN：0304-3835
通讯作者：
Cao, Deliang;Liao, Duan-Fang
作者机构：
[Huang, Chenfei; Shen, Yi; Cao, Yu; Bu, Yiwen; Cao, Deliang] Southern Illinois Univ, Sch Med, Simmons Canc Inst, Dept Med Microbiol Immunol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.;[Cai, Guoshuai] Geisel Sch Med Dartmouth, Dept Genet, Hanover, NH 03755 USA.;[Zeng, Xi] Univ South China, Canc Res Inst, Hengyang 421001, Hunan, Peoples R China.;[Wang, Yuhong; Cao, Deliang; Huang, Dan; Liao, Duan-Fang; Cai, Chuan] Hunan Univ Chinese Med, State Key Lab Chinese Med Powder & Med Innovat Hu, Div Stem Cell Regulat & Applicat, Changsha 410208, Hunan, Peoples R China.;[Liao, Duan-Fang] Hunan Univ Tradit Chinese Med, Div Stem Cell Regulat & Applicat, 1 Xiangzui Rd, Changsha 410208, Hunan, Peoples R China.
通讯机构：
[Cao, Deliang] S;[Liao, Duan-Fang] H;Southern Illinois Univ, Sch Med, Simmons Canc Inst, Dept Med Microbiol Immunol & Cell Biol, 913 N Rutledge St, Springfield, IL 62794 USA.;Hunan Univ Tradit Chinese Med, Div Stem Cell Regulat & Applicat, 1 Xiangzui Rd, Changsha 410208, Hunan, Peoples R China.
关键词：
NF-kappa B RelA/p65;ABCC6;RITA;Chemoresistance
摘要：
Inactivation of p53 occurs frequently in various cancers. RITA is a promising anticancer small molecule that dissociates p53-MDM2 interaction, reactivates p53 and induces exclusive apoptosis in cancer cells, but acquired RITA resistance remains a major drawback. This study found that the site-differential phosphorylation of nuclear factor-kappa B (NF-kappa B) RelA/p65 creates a barcode for RITA chemosensitivity in cancer cells. In naive MCF7 and HCT116 cells where RITA triggered vast apoptosis, phosphorylation of RelA/p65 increased at Ser536, but decreased at Ser276 and Ser468; oppositely, in RITA-resistant cells, RelA/p65 phosphorylation decreased at Ser536, but increased at Ser276 and Ser468. A phosphomimetic mutation at Ser536 (p65/S536D) or silencing of endogenous RelA/p65 resensitized the RITA-resistant cells to RITA while the phosphomimetic mutant at Ser276 (p65/S276D) led to RITA resistance of naive cells. In mouse xenografts, intratumoral delivery of the phosphomimetic p65/S536D mutant increased the antitumor activity of RITA. Furthermore, in the RITA-resistant cells ATP-binding cassette transporter ABCC6 was upregulated, and silencing of ABCC6 expression in these cells restored RITA sensitivity. In the naive cells, ABCC6 delivery led to RITA resistance and blockage of p65/S536D mutant-induced RITA sensitivity. Taken together, these data suggest that the site-differential phosphorylation of RelA/p65 modulates RITA sensitivity in cancer cells, which may provide an avenue to manipulate RITA resistance. (C) 2016 Elsevier Ireland Ltd. All rights reserved.
摘要：
Low shear stress plays a crucial role in the initiation and progression of atherosclerotic lesions. However, the detailed mechanisms of these processes remain unclear. In this study, the effect of low shear stress on endothelial cell autophagy and its potential mechanism were investigated. Results showed autophagy dysfunction and ten-eleven translocation 2 (TET2) protein downregulation during atherosclerotic lesion progression. Autophagic markers BECLIN 1 and LC3II/LC3I under low shear stress (5 dyne/cm(2)) obviously decreased compared with those under physiological shear stress (15 dyne/cm(2)), whereas autophagic substrate p62 increased. TET2 expression was also downregulated under low shear stress. Endothelial cell autophagy was improved with TET2 overexpression but was impaired by TET2 siRNA treatment. Moreover, TET2 overexpression upregulated the expression of endothelial cell nitric oxide synthase (eNOS) and downregulated the expression of endothelin-1 (ET-1). TET2 siRNA further attenuated eNOS expression and stimulated ET-1 expression. Overall, the results showed that low shear stress downregulated endothelial cell autophagy by impaired TET2 expression, which might contribute to the atherogenic process.
摘要：
Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-kappaB) signaling plays a dominant role in the pathogenesis of liver ischemia-reperfusion (IR) injury. Dexmedetomidine (Dex) protects the liver against IR injury via alpha(2)-adrenoceptor activation, but the contribution of TLR4 signaling remains unknown. The authors aimed to examine whether pretreatment with Dex produces hepatic protection and investigate the influence of Dex on TLR4/NF-kappaB signaling. Dex was given via intraperitoneal injection 30 min prior to orthotopic autologous liver transplantation (OALT) in rats, and three alpha(2)-adrenoceptor antagonists including atipamezole (a nonselective alpha(2) receptor blocker), ARC-239 (a specific alpha2B/C blocker) and BRL-44408 (a specific alpha2A blocker) were injected intraperitoneally 10 min before Dex administration. Histopathologic evaluation of the liver and the measurement of serum alanine aminotransferase activity, TLR4/NF-kappaB expression in the liver, and pro-inflammatory factors (serum tumor necrosis factor-alpha, interleukin-1beta and hepatic myeloperoxidase) concentrations were performed 8 h after OALT. Dex ameliorated liver injury after OALT probably by suppressing the TLR4/NF-kappaB pathway and decreasing inflammatory mediator levels. The protective effects of Dex were reversed by atipamezole and BRL-44408, but not by ARC-239, suggesting that these effects were mediated in part by the alpha2A subtype. In conclusion, Dex attenuates liver injury partly via the alpha2A-adrenoceptor subtype, and the mechanism is due to the suppression of the TLR4/NF-kappaB pathway.
摘要：
BACKGROUND: To investigate whether pretreatment with dexmedetomidine (Dex) has a protective effect against acute lung injury (ALI) in an orthotopic autologous liver transplantation (OALT) rat model and to explore the mechanisms responsible for the protective effect of Dex against lung injury. METHODS: Forty-eight rats underwent OALT and were randomly divided into six groups (n = 8 in each group) that received 10 microg/kg Dex, 50 microg/kg Dex, 50 microg/kg Dex + nonspecific alpha2-adrenergic receptor (AR) antagonist atipamezole, 50 microg/kg Dex + specific alpha2B/C-AR antagonist ARC-239, 50 microg/kg Dex + specific alpha2A-AR antagonist BRL-44408, or the same amount of normal saline. The sham rats (n = 8) underwent anesthesia induction, laparotomy, and separation of the portal vein without liver ischemia and reperfusion. Lung tissue sections were stained with hematoxylin and eosin (HE) to visualize the damage. The expression of Toll-like receptor 4 (TLR4) and the phospho-nuclear factor (NF)-kappaB p65 subunit as well as inflammatory cytokines was measured. RESULTS: Rats exhibited increased histological lung injury scores and pulmonary edema following OALT. Pretreatment with 50 mug/kg Dex attenuated OALT-induced lung injury in rats, probably by inhibiting the activation of the TLR4-NF-kappaB signaling pathway. The protective effect of Dex could be blocked by atipamezole or BRL-44408, but not by ARC-239, suggesting these effects of Dex were mediated, at least in part, by the alpha2A-AR. CONCLUSIONS: Dex exerts protective effects against ALI following OALT, and this protection is associated with the suppression of TLR4-NF-kappaB signaling. Thus, pretreatment with Dex may be a useful method for reducing lung damage caused by liver transplantation.
作者机构：
[Wang, Yi; Ma, Ai-Jing; Luo, Li-Juan; Ye, Chang-Yun; Liu, Kai; Jin, Dong; Wang, Yan] Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, Beijing 102206, Peoples R China.;[Wang, Yi; Ma, Ai-Jing; Luo, Li-Juan; Ye, Chang-Yun; Liu, Kai; Jin, Dong; Wang, Yan] Collaborat Innovat Ctr Diag & Treatment Infect Di, Hangzhou, Zhejiang, Peoples R China.;[Li, Dong-Xun] Guiyang Med Univ, Dept Microbiol, Guiyang 550004, Guizhou, Peoples R China.;[Liu, Dong-Xin] Univ South China, Pathogen Biol Inst, Hengyang 421000, Hunan, Peoples R China.;[Ye, Chang-Yun] Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, Changbai Rd 155, Beijing 102206, Peoples R China.
通讯机构：
[Ye, Chang-Yun] C;Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, Changbai Rd 155, Beijing 102206, Peoples R China.
摘要：
We have devised a novel amplification strategy based on isothermal strand-displacement polymerization reaction, which was termed multiple cross displacement amplification (MCDA). The approach employed a set of ten specially designed primers spanning ten distinct regions of target sequence and was preceded at a constant temperature (61-65 degrees C). At the assay temperature, the double-stranded DNAs were at dynamic reaction environment of primer-template hybrid, thus the high concentration of primers annealed to the template strands without a denaturing step to initiate the synthesis. For the subsequent isothermal amplification step, a series of primer binding and extension events yielded several single-stranded DNAs and single-stranded single stem-loop DNA structures. Then, these DNA products enabled the strand-displacement reaction to enter into the exponential amplification. Three mainstream methods, including colorimetric indicators, agarose gel electrophoresis and real-time turbidity, were selected for monitoring the MCDA reaction. Moreover, the practical application of the MCDA assay was successfully evaluated by detecting the target pathogen nucleic acid in pork samples, which offered advantages on quick results, modest equipment requirements, easiness in operation, and high specificity and sensitivity. Here we expounded the basic MCDA mechanism and also provided details on an alternative (Single-MCDA assay, S-MCDA) to MCDA technique.
作者机构：
[Huang, Pinjie; Cai, Jun; Chi, Xinjin; Wu, Shan; Yao, Hui; Wang, Yiheng] Sun Yat Sen Univ, Dept Anesthesiol, Affiliated Hosp 3, Guangzhou 510630, Guangdong, Peoples R China.;[Jin, Yi] Sun Yat Sen Univ, Dept Pathol, Affiliated Hosp 3, Guangzhou 510630, Guangdong, Peoples R China.;[Wang, Yiheng] Univ South China, Dept Anesthesiol, Affiliated Hosp 1, Hengyang 421001, Peoples R China.;[Xia, Zhengyuan] Univ Hong Kong, Dept Anesthesiol, Li Ka Shing Fac Med, Hong Kong, Hong Kong, Peoples R China.
通讯机构：
[Chi, Xinjin] S;Sun Yat Sen Univ, Dept Anesthesiol, Affiliated Hosp 3, Guangzhou 510630, Guangdong, Peoples R China.
摘要：
Patients who undergo orthotopic liver transplantation often sustain acute kidney injury(AKI). The toll-like receptor 4(TLR4)/Nuclear factor-кB(NF-кB) pathway plays a role in AKI. Dexmedetomidine(Dex) has been shown to attenuate AKI. The current study aimed to determine whether liver transplantation-induced AKI is associated with inflammatory response, and to assess the effects of dexmedetomidine pretreatment on kidneys in rats following orthotopic autologous liver transplantation(OALT). Seventy-seven adult male rats were randomized into 11 groups. Kidney tissue histopathology and levels of blood urea nitrogen(BUN) and serum creatinine(SCr) were evaluated. Levels of TLR4, NF-κB, tumor necrosis factor-α, and interleukin-1β levels were measured in kidney tissues. OALT resulted in significant kidney functional impairment and tissue injury. Pre-treatment with dexmedetomidine decreased BUN and SCr levels and reduced kidney pathological injury, TLR4 expression, translocation of NF-κB, and cytokine production. The effects of dexmedetomidine were reversed by pre-treatment with atipamezole and BRL44408, but not ARC239. These results were confirmed by using α2A-adrenergic receptor siRNA which reversed the protective effect of dexmedetomidine on attenuating NRK-52E cells injury induced by hypoxia reoxygenation. In conclusion, Dexmedetomidine-pretreatment attenuates OALT-induced AKI in rats which may be contributable to its inhibition of TLR4/MyD88/NF-κB pathway activation. The renoprotective effects are related to α2A-adrenergic receptor subtypes.
摘要：
Aims: Induction of heme oxygenase-1 (HO-1) has been widely accepted to be neuro-protective. This study aimed to examine whether hemin (a HO-1 inducer) attenuates neuronal damage in the hippocampus induced by orthotopic autologous liver transplantation (OALT) in adult rats. Main methods: Rats were randomly allocated into four groups (n = 8 each): (i) Sham control group; (ii) OALT model group; (iii) Hemin + OALT group, with intra-peritoneal (i.p.) injection of hemin (5 mg/kg) 24 hours (h) before the OALT; and (iv) ZnPP (a HO-1 inhibitor) + OALT group, with i.p. injection of ZnPP (32 mg/kg) 24 h before the OALT. Twenty four hours after the surgery, the hippocampal tissues were collected for electron microscopic examination and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) analysis. The levels of hippocampal HO-1 protein and serum S-100 beta, the concentrations of regional tumor necrosis factor-alpha (TNF-alpha) and interleukins (IL-6, IL-10), as well as the status of malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) in the hippocampus were assessed. Key findings: Rats suffered severe neuronal damage in the hippocampus after OALT, mainly in apoptosis. Pretreatment with hemin obviously alleviated the damage; up-regulated the HO-1 protein level; inhibited the release of TNF-alpha, IL-6 and MDA; and promoted the activities of SOD, CAT and IL-10; however, pre-treatment with ZnPP did not exhibit the opposite effect, except that a marked increase in serum S-100 beta level was detected. Significance: Hemin up-regulated the expression of HO-1 and attenuated hippocampal neuronal damage induced by OALT. (C) 2015 Elsevier Inc. All rights reserved.
摘要：
UNLABELLED: Aeromonas hydrophila has been increasingly implicated as the aetiologic agent of various human diseases. Therefore, reliable laboratory detection and identification of this bacterium has become clinically and epidemiologically desirable. We developed a nearly instrument-free, simple molecular method for rapid detection of Aer. hydrophila using a cross-priming amplification (CPA) assay with the desA gene as the target. The desA gene is crucial for the survival and growth of Aer. hydrophila under iron starvation. The results can be visualized as colour changes without opening the reaction tubes. No false-positive results were observed for the 33 non-Aer. hydrophila strains tested to evaluate assay specificity. The limit of detection for Aer. hydrophila was approximately 200 copies of desA per reaction (on reference plasmids) and 5 x 10(3) CFU g(-1) Aer. hydrophila in simulated human stool, which is the same sensitivity as a qPCR assay. The performance of the CPA assay was also evaluated with 100 stool specimens from diarrhoea patients and 40 environmental water samples. In conclusion, the simplicity, cost-effectiveness and nearly instrument-free platform of the CPA assay make it practical for use in primary care facilities and smaller clinical laboratories. SIGNIFICANCE AND IMPACT OF THE STUDY: Aeromonas hydrophila is a human pathogen that infects via exposed wounds or ingestion of contaminated water and food. In this study, a CPA-based PCR method was developed for specific, rapid, cost-effective detection of Aer. hydrophila, and the test results could be visualized without opening the reaction tubes. This is the first report on the application of the CPA method for the detection of Aer. hydrophila. This novel method could be practical for use in primary care facilities and smaller clinical laboratories.
摘要：
FBXW7 gene (F-box and WD-40 domain protein 7) is also named HCDC4 and is a significant tumor suppressor gene, which can regulate human cell cycle, proliferation and differentiation. In this study, we tend to investigate protein expression and related biological functions of FBXW7 gene. FBXW7 expression level in renal cell carcinoma (RCC) tissues is highly related to its clinical pathologic grade (P = 0.0094) and TNM phase (P = 0.0080) and is highly lower than in paracancerous normal tissues through immunohistochemistry study. FBXW7 high-expression patients have overall better prognosis than low-expression patients (P < 0.001). After transfected with FBXW7 plasmid, the RCC cell lines ACHN and A704 showed a depressed proliferation activity and high proportion of apoptosis through CCK8, colony formation and flow cytometry assay studies. By Western blot analysis, expression of cell proliferation-activating protein c-Myc and c-Jun is downregulated in FBXW7 high-expression RCC compared with negative control. These data suggested that FBXW7 is a significant tumor suppressor gene in RCC.
