TBL1XR1 induces cell proliferation and inhibit cell apoptosis by the PI3K/AKT pathway in pancreatic ductal adenocarcinoma

BACKGROUND Pancreatic ductal adenocarcinoma(PDAC) is one of the deadliest solid tumors. Identification of diagnostic and therapeutic biomarkers for PDAC is urgently needed. Transducin(β)-like 1 X-linked receptor 1(TBL1 XR1) has been linked to the progression of various human cancers. Nevertheless, the function and role of TBL1 XR1 in pancreatic cancers are unclear.AIM To elucidate the function and potential mechanism of TBL1 XR1 in the development of PDAC.METHODS Ninety patients with histologically-confirmed PDAC were included in this study. PDAC tumor samples and cell lines were used to determine the expression of TBL1 XR1. CCK-8 assays and colony formation assays were carried out to assess PDAC cell viability. Flow cytometry was performed to measure the changes in the cell cycle and cell apoptosis. Changes in related protein expression were measured by western blot analysis. Animal analysis was conducted to confirm the impact of TBL1 XR1 in vivo.RESULTS Patients with TBL1 XR1-positive tumors had worse overall survival than those with TBL1 XR1-negative tumors. Moreover, we found that TBL1 XR1 strongly promoted PDAC cell proliferation and inhibited PDAC cell apoptosis. Moreover, knockdown of TBL1 XR1 induced G0/G1 phase arrest. In vivo animal studies confirmed that TBL1 XR1 accelerated tumor cell growth. The results of western blot analysis showed that TBL1 XR1 might play a key role in regulating PDAC cell proliferation and apoptosis via the PI3 K/AKT pathway.CONCLUSION TBL1 XR1 promoted PDAC cell progression and might be an effective diagnostic and therapeutic marker for pancreatic cancer.     

AKT pathway in neuroblastoma and its therapeutic implication

Neuroblastoma is a frequent pediatric tumor with a poor outcome in spite of aggressive treatment, even with autologous hematopoietic stem cell transplantation. The overall cure rate of 40% is unsatisfactory and new therapeutic strategies are urgently needed. AKT is a major mediator of survival signals that protect cells from apoptosis and regulate cell proliferation. The AKT signaling network is considered a key determinant of the biological aggressiveness of these tumors. In this article, the authors discuss the relation between activators of AKT in neuroblastoma, in particular, growth factors such as IGF-1, TRK, GDNF, VEGF and EGF, and their effects on tumoral proliferation, differentiation and apoptosis. Numerous other proteins interact with AKT in neuroblastoma. Several are relatively well characterized, such as PTEN and retinoic acid; others are new and potentially interesting, such as PKC and anaplastic lymphoma kinase. Specific inhibition of AKT has been studied, such as with LY249002, with significant effects on cell progression and apoptosis in tumoral cells. Moreover, a series of new drugs, such as geldanamycin and rapamycin, directly modify the expression of AKT in tumoral cells. Few specific inhibitors of AKT are available; less specific inhibitors are probably unsuitable therapeutic options in neuroblastoma. Drugs with a direct or indirect inhibitory effect on the AKT pathway, used alone or in combination with other drugs, seem to hold great promise as a new therapeutic modality in neuroblastoma.     

间变性大细胞淋巴瘤中AKT、mTOR途径活化的研究

目的 研究间变性大细胞淋巴瘤(ALCL)患者中间变性淋巴瘤激酶(ALK)及磷酸化AKT(p-AKT)、mTOR(p-mTOR)、4E-BPI(p-4E-BPI)和p70S6K(p-p70S6K)的表达特点、临床意义及相互关系.方法 应用免疫组织化学EnVision法检测ALK蛋白及p-AKT、p-mTOR、p-4E-BP1、p-p70S6K蛋白的表达.结果 81例ALCL患者中有51例(63.0%)表达ALK蛋白,30例(37.0%)不表达,ALK阳性患者预后优于阴性患者(P<0.05).71例患者中54例(76.1%)表达p-AKT,p-AKT的表达与ALK表达相关(P<0.05);57例(80.3%)表达p-mTOR,p-mTOR的表达与ALK、p-AKT表达相关(P<0.05);64例(90.1%)表达p-4E-BP1,66例(93.0%)表达p-p70S6K,p-4E-BP1及p-p70S6K的表达与p-mTOR表达相关(P<0.05),与ALK、磷酸化p-AKT表达无关(P>0.05).p-AKT、P-mTOR、p-4E-BP1及p-p70S6K的表达与预后无关(P>0.05).COX比例风险回归分析表明ALK的表达、体质性症状对患者生存影响有统计学意义(P<0.05),其中,ALK的表达对生存的影响最大.结论 p-AKT、P-mTOR、p-4E-BP1和p-p70S6K在ALCL患者中均有表达,但在ALK阳性患者中表达率显著高于阴性患者.p-AKT、P-mTOR表达与ALK表达相关,提示在ALK阳性ALCL患者中存在AKT/mTOR通路的激活,但无明显的预后意义. 无关(P>0.05).p-AKT、P-mTOR、p-4E-BP1及p-p70S6K的表达与预后无关(P>0.05).COX比例风险回归分 表明ALK的表达、体质性症状对患者生存影响有统计学意义(P<0.05),其中,ALK的表达对生存的影响最大.结论 p-AKT、P-mTOR、p-4E-BP1和p-p70S6K在ALCL患者中均有表达,但在ALK阳性患者中表达率显著高于阴性患者.p-AKT、P-mTOR表达与ALK表达相关,提示在ALK阳性ALCL患者中存在AKT/mTOR通路的激活,但无明显的预后意义. 无关(P>0.05).     

胡桃醌对胃癌细胞耐药性及细胞中ANXA2、ERCC1表达和AKT/mTOR信号通路的影响

目的研究胡桃醌对胃癌细胞顺铂耐药性及细胞中膜联蛋白A2(ANXA2)和切除修复交叉互补基因1(ERCC1)表达,及对AKT/mTOR信号通路的影响。方法将BGC-823/DDP细胞株随机分为5组:对照组、胡桃醌组、顺铂组、胡桃醌联合顺铂组、LY294002(AKT抑制剂)联合顺铂组。采用MTT法和流式细胞术检测细胞活性和凋亡率,Western blotting检测细胞中相关蛋白表达。结果与对照组比较,顺铂组细胞凋亡率及细胞中Caspase-3、Caspase-9蛋白表达水平升高(P<0.05),而ANXA2、ERCC1、p-AKT、p-mTOR蛋白表达水平降低(P<0.05);与顺铂组比较,胡桃醌不同浓度组细胞中P-gp、MRP1蛋白表达水平降低(P<0.05),细胞增殖抑制率、细胞凋亡率及细胞中Caspase-3、Caspase-9蛋白表达水平升高(P<0.05),ANXA2、ERCC1、p-AKT、p-mTOR蛋白表达水平降低(P<0.05);与顺铂组比较,LY294002处理后,细胞增殖抑制率和凋亡率升高(P<0.05)。结论胡桃醌增强胃癌细胞对顺铂的敏感性,抑制细胞中ANXA2和ERCC1表达,抑制AKT/mTOR信号通路。     

氯沙坦对大鼠脑缺血再灌注损伤的保护作用及分子机制

目的研究氯沙坦对大鼠脑缺血再灌注(IR)损伤的保护作用及分子机制。方法 80只成年雄性SD大鼠随机分为假手术组、IR组、2. 5 mg/kg氯沙坦组、5. 0 mg/kg氯沙坦组、5. 0 mg/kg氯沙坦+LY组。2. 5 mg/kg氯沙坦组、5. 0 mg/kg氯沙坦组、5. 0 mg/kg氯沙坦+LY组在造模前连续给予氯沙坦灌胃干预14天,5. 0 mg/kg氯沙坦+LY组在造模前30 min给予磷脂酰肌醇3激酶(PI3K)抑制剂LY294002侧脑室注射。采用线栓法建立大鼠脑IR损伤模型,在再灌注后24 h评价神经功能缺损并测定脑组织含水量、细胞凋亡情况、凋亡相关基因表达水平。结果与假手术组比较,IR组大鼠的脑组织含水量、神经功能缺损评分、TUNEL阳性率及脑组织中Bcl-2相关X蛋白(Bax)/GAPDH、细胞色素C(Cyt C)/GAPDH、Cleaved-caspase-3/Pro-caspase-3的水平均显著增加,脑组织中B淋巴细胞瘤2蛋白(Bcl-2)/GAPDH、p-PI3K/PI3K、p-AKT/AKT的水平显著减少;与IR组比较,2. 5 mg/kg氯沙坦组和5. 0 mg/kg氯沙坦组大鼠的脑组织含水量、神经功能缺损评分、TUNEL阳性率及脑组织中Bax/GAPDH、Cyt C/GAPDH、Cleaved-caspase-3/Pro-caspase-3的水平均显著减少,脑组织中Bcl-2/GAPDH、p-PI3K/PI3K、p-AKT/AKT的水平显著增加;与5. 0 mg/kg氯沙坦组比较,5. 0 mg/kg氯沙坦+LY组大鼠的脑组织含水量、神经功能缺损评分、TUNEL阳性率及脑组织中Bax/GAPDH、Cyt C/GAPDH、Cleaved-caspase-3/Pro-caspase-3的水平均显著增加,脑组织中Bcl-2/GAPDH、p-PI3K/PI3K、p-AKT/AKT的水平显著减少。结论氯沙坦通过上调PI3K/AKT通路减轻大鼠脑IR损伤。     

S-allyl-l-cysteine attenuates bleomycin-induced pulmonary fibrosis and inflammation via AKT/NF-κB signaling pathway in mice

Idiopathic pulmonary fibrosis (IPF) is a progressive and lethal lung disease characterized by inflammation, multifocal fibrotic lesions and excessive collagen deposition with limited therapies. As a major bioactive compound in garlic, S-allyl-l-cysteine (SAC) is a neuroprotective drug candidate to prevent cognitive decline, however, its anti-pulmonary fibrotic activity remains unknown. Here, we investigated whether SAC could attenuate bleomycin (BLM)-induced pulmonary fibrosis and inflammation in mice. Our results showed that SAC dose-dependently reduced the infiltration of inflammatory cells, pulmonary lesions and collagen deposition in BLM treated mice with downregulated mRNA expression levels of fibrotic genes including alpha smooth muscle actin (α-SMA), fibronectin, collagen I and collagen III as well as the protein level of α-SMA. In addition, SAC could also reduce the mRNA expression of inflammatory mediators such as TNF-α and iNOS. Furthermore, higher phosphorylation of AKT and NF-κB p65 in IPF patient samples and murine samples was verified by immunohistochemistry while SAC could decrease the phosphorylation level of AKT and NF-κB p65 in mice stimulated with BLM. These findings, for the first time, indicate that SAC might mediate AKT/NF-κB signaling pathway to inhibit BLM-induced pulmonary fibrosis and support the potential role of SAC as an anti-pulmonary fibrosis agent.     

西黄丸组分中药调节肿瘤微环境中Treg细胞PI3K/AKT通路的抗肿瘤作用机制研究

目的 研究西黄丸组分中药是否与西黄丸具有相似的抗癌作用及机制。方法 4T1乳腺癌细胞荷瘤建立动物模型,西黄丸低、中、高剂量（0.39、0.78、1.95 g/kg）和西黄丸组分中药0.3 mL （74.5 mg/kg榄香烯、2.73 mg/kg牛磺酸、0.52mg/kg麝香酮和4.75 mg/kg 11-羰基-β乙酯乳香酸混合物,对应于西黄丸高剂量组） ig给药14 d,剥离肿瘤组织,称质量,匀浆,制备单细胞悬液,免疫磁硃分离Treg细胞。流式细胞术和免疫组化检测肿瘤微环境中Treg细胞数量变化情况,Western Blotting检测肿瘤微环境中Treg细胞磷脂酰肌醇3-激酶/蛋白激酶B （PI3K/AKT）蛋白表达情况,实时荧光定量PCR （qRT-PCR）检测肿瘤微环境中Treg细胞PI3K、AKT mRNA表达情况。结果 与模型组及西黄丸低、中剂量组比较,西黄丸组分中药组肿瘤质量显著减少（P<0.05）;流式细胞术和免疫组化结果显示,肿瘤微环境中Treg细胞数量显著减少（P<0.05）;Western Blotting和qRT-PCR结果显示,肿瘤微环境中Treg细胞PI3K、AKT蛋白及mRNA表达显著下降（P<0.05）。与西黄丸高剂量组比较,西黄丸组分中药组肿瘤质量、Treg细胞数量和PI3K、AKT蛋白及mRNA表达均明显升高（P<0.05）。结论 西黄丸组分中药通过抑制肿瘤微环境中Treg细胞PI3K/AKT信号通路的表达减少Treg细胞数量,进而表现与西黄丸相似的抗癌作用和机制。