CADM1在大肠癌细胞系中的表达及与启动子甲基化的关系

目的：探讨CADM1在大肠癌细胞系中的表达及与启动子甲基化的关系。方法用RT－PCR法检测大肠癌细胞系中 CADM1基因 mRNA 表达水平,Western blot 法检测细胞系 CADM1蛋白表达情况,BSP 法和 MSP法检测 CADM1基因启动子甲基化状态。结果CADM1 mRNA 在24％大肠癌细胞系表达缺失,在50％细胞系中CADM1 mRNA 表达减少。75％大肠癌细胞系检测到 CADM1基因启动子甲基化阳性,且表达缺失。结论CADM1基因启动子甲基化与大肠癌的发生发展有密切关系,该基因的甲基化检测可作为癌症早期筛选、监测预后的肿瘤标志物。     

AGE‐modified basement membrane cooperates with Endo180 to promote epithelial cell invasiveness and decrease prostate cancer survival

Biomechanical strain imposed by age‐related thickening of the basal lamina and augmented tissue stiffness in the prostate gland coincides with increased cancer risk. Here we hypothesized that the structural alterations in the basal lamina associated with age can induce mechanotransduction pathways in prostate epithelial cells (PECs) to promote invasiveness and cancer progression. To demonstrate this, we developed a 3D model of PEC acini in which thickening and stiffening of basal lamina matrix was induced by advanced glycation end‐product (AGE)‐dependent non‐enzymatic crosslinking of its major components, collagen IV and laminin. We used this model to demonstrate that antibody targeted blockade of CTLD2, the second of eight C‐type lectin‐like domains in Endo180 (CD280, CLEC13E, KIAA0709, MRC2, TEM9, uPARAP) that can recognize glycosylated collagens, reversed actinomyosin‐based contractility [myosin‐light chain‐2 (MLC2) phosphorylation], loss of cell polarity, loss of cell–cell junctions, luminal infiltration and basal invasion induced by AGE‐modified basal lamina matrix in PEC acini. Our in vitro results were concordant with luminal occlusion of acini in the prostate glands of adult Endo180^{ΔEx2–6/ΔEx2–6} mice, with constitutively exposed CTLD2 and decreased survival of men with early (non‐invasive) prostate cancer with high epithelial Endo180 expression and levels of AGE. These findings indicate that AGE‐dependent modification of the basal lamina induces invasive behaviour in non‐transformed PECs via a molecular mechanism linked to cancer progression. This study provides a rationale for targeting CTLD2 in Endo180 in prostate cancer and other pathologies in which increased basal lamina thickness and tissue stiffness are driving factors. Copyright © 2014 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

健康人心率变异时域法正常值分析

为了解国人心率变异(HRV)时域法指标正常值,检测1524例17—94岁健康人24h动态心电图,分析HRV时域法5项指标。结果显示:(1)SDNN、SDANN、SDNN_(Index),rMSSD和PNN_(50)均值分别为127±33、116±32、49±14、29±12(ms)和8±9％,与国内研究一致。(2)HRV时域法各指标随年龄增加而降低(r=0.30——0.48,P<0.01),中青年组与老年组HRV差异有显著意义(P<0.01)。(3)男性SDNN、SDANN和SDNN_Index大于女性(P<0.05),而rMSSD和PNN_(50)小于女性(P<0.05)。SnNN、SDANN、SDNN_(Index)和rMSSD单侧下限值分别为73、64、26和9ms,PNN_(50)呈偏态分布,临床意义有待探讨。     

Plasminogen activator induction and platelet aggregation by phorbol and some of its derivatives: Correlation with skin irritancy and tumor-promoting activity

Summary Phorbol and eight of its derivatives were investigated for their ability to stimulate the synthesis of the enzyme plasminogen activator in cultured chick embryo fibroblasts and to aggregate human blood platelets and have been assayed for tumor, promoting and skin, irritant activities. Over a range of concentrations, elevation in the levels of plasminogen activator activity induced by phorbol derivatives correlates well with their promoting and irritant properties. In the platelet aggregation assay however, the parallelism between the activities measured in different biological assays was less complete. While strong promoters, such as TPA, are potent aggregating agents, and weak promoters, such as PDA, are poor or ineffective inducers of aggregation, two derivatives, PDD and PDB, deviate from this general result. Platelets must be exposed to PDD in relatively high concentrations before they will aggregate, and PDB was found to be the most potent aggregating agent of all the derivatives tested.

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Abbreviations TPA 12-O-tetradecanoylphorbol-13-acetate - PDD phorbol-12,13-didecanoate - TPA--oxide 12-O-tetradecanoylphorbol-13-acetate-6,7-oxide - PDB phorbol-12,13-dibenzoate - PDA phorbol-12,13-diacetate - 4-OMe-TPA 4-O-methyl-12-O-tetradecanoylphorbol-13-acetate - 1,2-dihydro-TPA--oxide 1,2-dihydro-12-O-tetradecanoylphorbol-13-acetate-6,7-oxide - 4-PDD 4-phorbol-12,13-didecanoate This work was supported by a Grant-in-Aid from the American Heart Association and with funds contributed in part by the Suffolk Heart Association (Grant 78 747), the National Cancer Institute (Grants CA08290 and CA00118), and the American Cancer Society (Grant PDT1)     

作用于雄激素受体不同结合位点的前列腺癌治疗药物的研究进展

前列腺癌是最常见的男性泌尿生殖系统的恶性肿瘤。雄激素受体在前列腺癌的发生、发展中起着重要作用。目前,所有治疗前列腺癌的药物(包括第一代的氟他胺、比卡鲁胺、尼鲁米特和第二代的恩扎鲁胺)都与雄激素受体的配体结合口袋结合,并且这些药物有着相似的分子结构,这可能引起药物之间的交叉耐药。为了避免耐药性的产生,研究者们致力于发现雄激素受体上新的药物结合位点。除雄激素受体配体结合位点外,主要对作用于氮端结合位点上的第一活性功能区(AF1)、第二活性功能区(AF2)、AF2附近的第三结合功能区(BF3)和DNA结合位点(DBD)的药物进行综述。     

Muscarinic modulation of TREK currents in mouse sympathetic superior cervical ganglion neurons

Muscarinic receptors play a key role in the control of neurotransmission in the autonomic ganglia, which has mainly been ascribed to the regulation of potassium M‐currents and voltage‐dependent calcium currents. Muscarinic agonists provoke depolarization of the membrane potential and a reduction in spike frequency adaptation in postganglionic neurons, effects that may be explained by M‐current inhibition. Here, we report the presence of a riluzole‐activated current (I_RIL) that flows through the TREK‐2 channels, and that is also inhibited by muscarinic agonists in neurons of the mouse superior cervical ganglion (mSCG). The muscarinic agonist oxotremorine‐M (Oxo‐M) inhibited the I_RIL by 50%, an effect that was abolished by pretreatment with atropine or pirenzepine, but was unaffected in the presence of himbacine. Moreover, these antagonists had similar effects on single‐channel TREK‐2 currents. I_RIL inhibition was unaffected by pretreatment with pertussis toxin. The protein kinase C blocker bisindolylmaleimide did not have an effect, and neither did the inositol triphosphate antagonist 2‐aminoethoxydiphenylborane. Nevertheless, the I_RIL was markedly attenuated by the phospholipase C (PLC) inhibitor ET‐18‐OCH3. Finally, the phosphatidylinositol‐3‐kinase/phosphatidylinositol‐4‐kinase inhibitor wortmannin strongly attenuated the I_RIL, whereas blocking phosphatidylinositol 4,5‐bisphosphate (PIP₂) depletion consistently prevented I_RIL inhibition by Oxo‐M. These results demonstrate that TREK‐2 currents in mSCG neurons are inhibited by muscarinic agonists that activate M1 muscarinic receptors, reducing PIP₂ levels via a PLC‐dependent pathway. The similarities between the signaling pathways regulating the I_RIL and the M‐current in the same neurons reflect an important role of this new pathway in the control of autonomic ganglia excitability.     

靶向p21基因saRNA抑制肝癌细胞生长实验研究

目的 探讨RNA激活p21对肝癌HepG2、Hep3b和SMMC-7721细胞生长和侵袭力的影响.方法 化学合成靶向p21的saRNA、阴性对照dsRNA,将其转染肝癌细胞系HepG2、Hep3b和SMMC-7721.每个细胞系分为3组,分别为p21-322组、阴性对照组和空白对照组,每组复3孔;p21-322组和阴性对照组分别采用p21-322 saRNA和阴性对照dsRNA进行转染,空白对照组不干预.转染后72 h,采用RT-PCR和Western blot检测各组细胞中的p21 mRNA和P21蛋白表达水平,采用MTT法检测细胞生长情况及划痕实验观察细胞侵袭能力的变化.结果 HepG2、Hep3b和SMMC-7721细胞p21-322组p21 mRNA相对表达水平分别为23.43±2.29、16.87±1.61、31.77±5.06,P21蛋白相对表达水平分别为55.93±12.66、32.91±5.17、24.96±6.81;空白对照组分别为3.53±0.07、2.39±0.02、5.70±0.89,3.21±0.03、2.91±0.14、4.15±0.12;阴性对照组分别为3.87±0.97、2.57±0.71、5.87±1.73,3.11±0.70、3.01±0.97、5.13±2.14;p21-322组p21 mRNA和蛋白相对表达水平明显高于空白对照组和阴性对照组(P＜0.01),空白对照组与阴性对照组比较差异无统计学意义(P＞0.05);细胞转染后第6天时HepG2、Hep3b、SMMC-7721细胞p21-322转染组细胞平均生长抑制率分别为41％、48％和52％;HepG2细胞p21-322组24 h后空白区域占原划痕区域面积的百分比((76±11)％)明显高于空白对照组((13±6)％)和阴性对照组((17±8)％)(P＜0.01),阴性对照组与空白对照组比较差异无统计学意义(P＞0.05).结论 靶向p21的RNAa能抑制肝癌细胞的生长和侵袭力,p21可作为一个具有肝癌治疗应用价值的靶基因.     

Intersectin1‐s,A multidomain adapter protein,Is essential for malignant glioma proliferation

Glioblastomas, the most aggressive form of primary brain tumors with a tendency to invade surrounding healthy brain tissues, remains an incurable disease. Intersectin (ITSN) is a multidomain adapter protein implicated in endocytosis, exocytosis, and multiple signaling pathways. Prior research of ours has shown intersectin1‐S (ITSN1‐S) is critical for the migration and invasion of glioma cells by regulating several key proteins. In this study, we established ITSN1‐S expression patterns in human tumor tissues. We discovered that ITSN1‐S expression was positively correlated with histological grade of gliomas and with poor patient prognosis. We also found that the expression of ITSN1‐S protein was essential to glioblastoma cell proliferation. Furthermore, through a series of expression constructs encoding different ITSN1‐S domains, we identified the critical roles of ITSN1‐S SH3 domains in the regulation of cell proliferation. This study also demonstrates evidence suggesting that the regulation of ITSN1‐S on glioblastoma cells proliferation is through the Raf/MEK/ERK pathway. In conclusion, this study suggests critical roles of ITSN1‐S in malignant glioma proliferation, indicating a potential usage of ITSN1‐S in the therapeutic intervention as a novel molecular target. GLIA 2015;63:1595–1605