MiR-21通过PTEN/PI3K/AKT信号通路抑制胰腺癌细胞系生长侵袭能力的体外研究

目的 探讨MiR-21对胰腺癌细胞生长和侵袭的影响.方法 采用脂质体Lipofectamine 2 000将MiR-21 inhibitor和MiR-21 inhibitor阴性对照转染至胰腺癌细胞ASPC-1,qRT-PCR检测转染效果,细胞免疫荧光、Western blot检测转染后目的蛋白的表达,Transwell、划痕实验检测细胞迁移侵袭能力,MTT法检测细胞的增殖活性,流式细胞术检测细胞周期分布的改变.结果 成功构建稳定下调MiR-21的胰腺癌细胞亚系ASPC-1-MiR-21,荧光定量PCR结果显示MiR-21表达量明显下调,抑制MiR-21表达后ASPC细胞的侵袭和增殖能力均受到明显抑制;抑制MiR-21能明显增强PTEN蛋白和减低p-AKT蛋白表达水平,对AKT蛋白水平无明显影响.结论 抑制MiR-21表达可能通过调控PTEN/AKT通路抑制胰腺癌细胞AS-PC-1增殖和侵袭.     

Proteus syndrome review: molecular,clinical, and pathologic features

Proteus syndrome is caused by an activating AKT1 mutation (c.49G>A, p.Glu17Lys). Many variable features are possible in this mosaic disorder, including: (i) disproportionate, asymmetric, and distorting overgrowth; (ii) bone abnormalities different from those observed in other disorders; (iii) a characteristic cerebriform connective tissue nevus made up of highly collagenized connective tissue; (iv) epidermal nevi in early life, consisting of acanthosis and hyperkeratosis; (v) vascular malformations of the capillary, venous, or lymphatic types; (vi) dysregulated adipose tissue including lipomas, lipohypoplasia, fatty overgrowth, and localized fat deposits; (vii) other unusual features, including bullous lung alterations; specific neoplasms; a facial phenotype associated with intellectual disability and/or seizures, and/or brain malformations; and (viii) deep vein thrombosis, resulting in premature death. Concluding remarks address diagnostic criteria, natural history, management, psychosocial issues, and differential diagnosis.     

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。     

Mammalian target of rapamycin inhibition in hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death worldwide. It is associated with a poor prognosis and has limited treatment options. Sorafenib, a multi-targeted kinase inhibitor, is the only available systemic agent for treatment of HCC that improves overall survival for patients with advanced stage disease; unfortunately, an effective second-line agent for the treatment of progressive or sorafenib-resistant HCC has yet to be identified. This review focuses on components of the mammalian target of rapamycin (mTOR) pathway, its role in HCC pathogenesis, and dual mTOR inhibition as a therapeutic option with potential efficacy in advanced HCC. There are several important upstream and downstream signals in the mTOR pathway, and alternative tumor-promoting pathways are known to exist beyond mTORC1 inhibition in HCC. This review analyzes the relationships of the upstream and downstream regulators of mTORC1 and mTORC2 signaling; it also provides a comprehensive global picture of the interaction between mTORC1 and mTORC2 which demonstrates the pre-clinical relevance of the mTOR pathway in HCC pathogenesis and progression. Finally, it provides scientific rationale for dual mTORC1 and mTORC2 inhibition in the treatment of HCC. Clinical trials utilizing mTORC1 inhibitors and dual mTOR inhibitors in HCC are discussed as well. The mTOR pathway is comprised of two main components, mTORC1 and mTORC2; each has a unique role in the pathogenesis and progression of HCC. In phase III studies, mTORC1 inhibitors demonstrate anti-tumor activity in advanced HCC, but dual mTOR (mTORC1 and mTORC2) inhibition has greater therapeutic potential in HCC treatment which warrants further clinical investigation.     

miR-93-3p对高糖诱导的肾小管上皮细胞损伤及炎症因子分泌的影响

目的 微小miRNA-93-3p（miR-93-3p）对高糖诱导的肾小管上皮细胞损伤及炎症因子分泌的影响。方法 体外培养肾小管上皮细胞,分为NC组、HG组、anti miR NC组、anti miR-93-3p组、HG+ miR NC组、HG+miR-93-3p组。采用实时荧光定量聚合酶链反应（qRT-PCR）检测miR 93 3p的表达;酶联免疫吸附（ELISA）法检测肿瘤坏死因子 α（TNF-α）、白细胞介素6（IL-6）水平;甲基噻唑基四唑（MTT）检测细胞增殖能力;流式细胞术检测细胞凋亡率;蛋白免疫印迹法（Western blot）检测细胞周期蛋白1（CyclinD1）、活化的含半胱氨酸的天冬氨酸蛋白水解酶3（C caspase-3）、磷酸化 磷脂酰肌醇激酶（p-PI3K）、磷酸化 蛋白激酶 B（p-AKT）表达量。结果 与NC组相比,HG组miR 93 3p的表达水平显著降低（P＜0.05）,细胞存活率显著降低（P＜0.05）,CyclinD1蛋白表达水平显著降低（P＜0.05）,而细胞凋亡率显著升高（P＜0.05）,C caspase 3、p PI3K、p AKT蛋白表达水平显著升高（P＜0.05）,TNF α、IL 6水平显著升高（P＜0.05）;与anti miR NC组相比,anti miR-93-3p组细胞存活率显著降低（P＜0.05）,细胞凋亡率显著升高（P＜0.05）,C caspase 3蛋白表达水平显著升高（P＜0.05）,TNF α、IL 6水平显著升高（P＜0.05）;与HG+ miR NC组相比,HG+miR 93 3p组细胞存活率显著升高（P＜0.05）,细胞凋亡率显著降低（P＜0.05）,CyclinD1蛋白表达水平显著升高（P＜0.05）,C caspase 3、p PI3K、p AKT蛋白表达水平显著降低（P＜0.05）,TNF α、IL 6水平显著降低（P＜0.05）。结论 miR 93 3p通过调控PI3K/AKT信号通路抑制高糖诱导的肾小管上皮细胞损伤及炎症因子分泌。     

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。     

AKT2、PTEN在胃癌组织中的表达及临床意义

目的探讨癌基因AKT2、磷酸酶基因(PTEN)在胃癌组织中的表达及临床意义。方法采用免疫组化法检测胃癌组织、癌旁正常组织以及萎缩性胃炎组织中AKT2和PTEN的表达。结果胃癌组织与萎缩性胃炎组织中AKT-2阳性率高于癌旁正常组织(P<0.01);胃癌组织中AKT-2阳性率明显高于萎缩性胃炎组织(P<0.05)。胃癌组织与萎缩性胃炎组织中PTEN阳性率低于癌旁正常组织(P<0.01);胃癌组织中PTEN阳性率亦低于萎缩性胃炎组织(P<0.01)。AKT2、PTEN的表达与肿瘤分化程度、有无淋巴转移及临床分期具有相关性(P<0.05);胃癌组织中PTEN和AKT2蛋白表达强度呈明显负相关(r=-0.349,P<0.05)。结论联合检测AKT2和PTEN蛋白的表达可作为判断胃癌生物学行为的辅助指标。     

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。     

Early pregnancy induced expression of prostaglandin E2 receptors EP2 and EP4 in the ovine endometrium and regulated by interferon tau through multiple cell signaling pathways

Prostaglandin E2 (PGE₂) plays pleiotropic roles at fetal-maternal interface during establishment of pregnancy. The objectives of the study were to: (i) determine regulation of PGE2 receptors EP1, EP2, EP3, and EP4 in the endometrium during the estrous cycle and early pregnancy; and (ii) understand endometrial epithelial and stromal cell-specific hormonal regulation of EP2 and EP4 in sheep. Results indicate that: (i) early pregnancy induces expression of EP2 and EP4 but not EP1 and EP3 proteins in the endometrium on days 12-16 compared to that of estrous cycle; (ii) intrauterine infusion of interferon tau (IFNT) increases expression of EP2 and EP4 proteins in endometrium; and (iii) IFNT activates distinct epithelial and stromal cell-specific JAK, EGFR, ERK1/2, AKT, or JNK signaling module to regulate expression of EP2 and EP4 proteins in the ovine endometrium. Our results indicate a role for EP2 and EP4-mediated PGE₂ signaling in endometrial functions and establishment of pregnancy in ruminants.