磷脂酰肌醇-3-激酶在结直肠癌组织中的表达及意义

目的:通过检测结直肠腺癌组织中磷酯酰肌醇-3-激酶(PI3K)蛋白的表达情况,探讨其表达特点及临床意义。方法:采用免疫组织化学光波/微波(Lightwave/Microwave,LW/MW)-EliVisionTM法检测100例结直肠癌、30例正常黏膜组织中PI3K的表达情况,并分析其与结直肠癌临床病理特点的关系。结果:PI3K蛋白在正常黏膜及结直肠癌组织中表达阳性率分别为6.67%(2/30)、56.00%(56/100),差异有统计学意义(P0.01)。PI3K蛋白表达与结直肠癌TNM分期、浆膜浸润和淋巴结转移相关(P0.05);而与年龄、性别、肿瘤大小、分化程度等指标不相关(P0.05)。结论:PI3K过度激活在结直肠癌的发生、发展、侵袭、转移中起着重要作用,这为其进行药物治疗及分子靶向治疗提供了新的着眼点。     

肾透明细胞癌中PTEN/PI3K/AKT信号通路的活性及其临床意义

目的 探讨人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)/磷脂酰肌醇3-激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(AKT)信号通路在肾透明细胞癌中的活性及其与临床特征之间的关系.方法 收集36对肾透明细胞癌及其邻近非肿瘤肾组织,Western blot检测AKT(p-S473)和PTEN蛋白表达,分析两者表达水平的相关性以及PTEN与肿瘤临床特征之间的关系.结果 非肿瘤肾组织中AKT(p-Ser473)(0.945±0.314),PTEN(3.611±0.947).与之比较,肾透明细胞癌中PTEN表达量下降(1.975±0.648),AKT(p-S473)增加(2.751±0.832),差异有统计学意义(P＜0.05).PTEN的表达水平与肿瘤临床特征之间无明显相关(P＞0.05).结论 肾透明细胞癌中PTEN可以调节PDK/AKT信号通路的活性,同时PTEN蛋白升高与AKT活性增加同时存在,提示在肾透明细胞癌中还存在使VFEN功能下降或促进AKT活性增加的因素.     

肾透明细胞癌中PTEN/PI3K/AKT信号通路的活性及其临床意义

目的 探讨人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)/磷脂酰肌醇3-激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(AKT)信号通路在肾透明细胞癌中的活性及其与临床特征之间的关系.方法 收集36对肾透明细胞癌及其邻近非肿瘤肾组织,Western blot检测AKT(p-S473)和PTEN蛋白表达,分析两者表达水平的相关性以及PTEN与肿瘤临床特征之间的关系.结果 非肿瘤肾组织中AKT(p-Ser473)(0.945±0.314),PTEN(3.611±0.947).与之比较,肾透明细胞癌中PTEN表达量下降(1.975±0.648),AKT(p-S473)增加(2.751±0.832),差异有统计学意义(P＜0.05).PTEN的表达水平与肿瘤临床特征之间无明显相关(P＞0.05).结论 肾透明细胞癌中PTEN可以调节PDK/AKT信号通路的活性,同时PTEN蛋白升高与AKT活性增加同时存在,提示在肾透明细胞癌中还存在使VFEN功能下降或促进AKT活性增加的因素.

Abstract：

Objective To discuss the activity of the phosphatase and tensin homologue deleted on chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/AKT pathway and the clinical significance of PTEN protein expression in clear-cell renal-cell carcinoma( CCRCC) tissues. Methods The protein of PTEN and AKT( phosphorylation at Ser473) of 36 paired CCRCC and adjacent non-neoplastic renal samples were analysed by Western blotting. Results The significantly increased AKT (phosphorylation at Ser473 ) with decreased PTEN protein were observed in CCRCC tissues compared with the adjacent non-neoplastic renal samples while the PTEN protein expression has no significant association with pathological parameters. Conclusion Our findings indicated that as PTEN dominantly inhibits AKT activation, the coexistence of high levels of the PTEN protein with enhanced AKT activation suggests the existence of novel mechanisms which attenuate PTEN function in CCRCC. These mechanisms may reduce PTEN function or increase AKT activation.     

肾透明细胞癌中PTEN/PI3K/AKT信号通路的活性及其临床意义

目的 探讨人第10号染色体缺失的磷酸酶及张力蛋白同源的基因(PTEN)/磷脂酰肌醇3-激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(AKT)信号通路在肾透明细胞癌中的活性及其与临床特征之间的关系.方法 收集36对肾透明细胞癌及其邻近非肿瘤肾组织,Western blot检测AKT(p-S473)和PTEN蛋白表达,分析两者表达水平的相关性以及PTEN与肿瘤临床特征之间的关系.结果 非肿瘤肾组织中AKT(p-Ser473)(0.945±0.314),PTEN(3.611±0.947).与之比较,肾透明细胞癌中PTEN表达量下降(1.975±0.648),AKT(p-S473)增加(2.751±0.832),差异有统计学意义(P＜0.05).PTEN的表达水平与肿瘤临床特征之间无明显相关(P＞0.05).结论 肾透明细胞癌中PTEN可以调节PDK/AKT信号通路的活性,同时PTEN蛋白升高与AKT活性增加同时存在,提示在肾透明细胞癌中还存在使VFEN功能下降或促进AKT活性增加的因素.     

Targeted therapy in colorectal carcinoma: more than a theory

Objective The aim of this review was to examine clinical trial data reporting the use of targeted therapies in colorectal cancer. Method Candidate trials were identified by a comprehensive literature search. Results The data on the use of targeted therapies; usually combined with chemotherapy in the treatment of colorectal cancer is accumulating rapidly. These new agents will increasingly become incorporated into standard treatment schedules. Conclusion Targeted therapy has moved rapidly from the laboratory to the clinic and is opening up potentially new and exciting areas for the development of the systemic treatment of colorectal cancer.     

PI3K/AKT/mTOR信号通路在肾上腺皮质癌组织的表达特点

目的分析PI3K/AKT/mTOR信号通路在肾上腺皮质癌组织中的表达特点及其意义。方法选取2008年1月至2012年12月经手术治疗且具有完整临床、病理资料的肾上腺皮质癌标本15例,以正常肾上腺组织(肾癌患者施行手术时所取下)12例作为对照。研究这些组织中该信号通路关键蛋白p-AKT、p-mTOR、p-S6以及血管内皮生长因子(VEGF)的表达情况。结果与对照组比较,p-AKT(12/15,80%)、p-mTOR(11/15,73.3%)、p-S6(13/15,86.7%)以及VEGF(12/15,80%)在肾上腺皮质癌中明显高表达。结论 PI3K/AKT/mTOR信号通路关键蛋白p-AKT、p-mTOR、p-S6以及VEGF参与肾上腺皮质癌的发生发展。     

VEGF-A及PI3K/AKT通路在结直肠癌中的表达及临床意义

目的 研究血管内皮生长因子（VEGF）-A及磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）通路在结直肠癌中的表达及临床意义。方法 选取我院2013年9月到2015年9月间收治的结直肠癌患者80例,作为观察组,同时切取患者癌旁正常肠黏膜组织作为对照组。分别采用免疫组化法检测癌组织和癌旁组织的VEGF-A及PI3K及 AKT表达水平,并分别统计其阳性表达率,比较两组间差异。同时单因素分析结直肠癌组织中VEGF-A、PI3K及AKT表达水平与患者临床病理资料相关性,同时Spearman相关性分析VEGF-A与PI3K及AKT表达相关性。术后随访5年,比较VEGF-A、PI3K及AKT阳性和阴性表达患者生存率。结果 观察组的VEGF-A及PI3K及AKT表达阳性率均明显高于对照组（P<0.05）。结直肠癌患者的VEGF-A及PI3K及AKT表达水平与性别、年龄、肿瘤直径、肿瘤部位等临床资料无明显相关性（P>0.05）,与TNM分期、分化程度、淋巴结转移及浸润深度明显相关（P<0.05）。结直肠癌组织中的VEGF-A表达与PI3K、AKT表达呈正相关（r=0.625, 0.517; 均P<0.05）。结直肠癌VEGF-A及PI3K及AKT阳性表达患者的5年生存率均低于阴性组（P<0.05）。结论 结直肠癌组织中的VEGF-A及PI3K及 AKT均存在高表达现象,且与肿瘤分期、淋巴结转移、分化及浸润深度呈现明显相关性,同时可减少患者生存期,可能参与结直肠癌病情发生发展,对临床诊断及治疗具有重要意义。     

PI3K/PTEN/AKT/mTOR信号通道在胃癌有关组织中的表达及临床意义

目的 研究PI3K/PTEN/AKT/mTOR信号通道在胃癌组织中的表达及临床意义.方法 采用免疫组化SP法分别检测33例胃癌及30例正常胃组织中mTOR、PTEN的表达情况,并分析二者与性别、年龄、淋巴结转移、浸润深度和分化程度的关系.结果 mTOR蛋白阳性反应产物主要分布于胞质,PTEN的蛋白阳性反应产物主要定位于胞核.将mTOR表达阴性,PTEN表达阳性为一组,与mTOR表达阳性而PTEN表达阴性为一组进行比较,发现对于不同的大体类型,组织学分型,二组间的差异有统计学意义(P<0.05),对于不同的淋巴结转移,临床病理分期,二组间的差异有统计学意义(P<0.01).结论 在PI3K/PTEN/AKT/mTOR信号通道中,mTOR与PTEN的联合表达可作为判断胃癌分期、判断预后的指标和化学治疗的靶点.     

PI3K/AKT/mTOR信号通路在肝癌细胞自体吞噬中的作用研究

目的探讨在肝癌细胞中PI3K／AKT／mTOR信号通路在氨基酸缺乏诱导的自体吞噬中的作用。方法观察HCCLM3、MHCC97L及SMMC-7721肝癌细胞在氨基酸缺乏情况下及与P13K抑制剂3-MA、Wortmannin及LY294002，mTOR抑制剂雷帕霉素协同作用下细胞活力变化。GFP—LC3荧光法观察细胞内自噬体的形成。Western印迹检测LC3-Ⅱ蛋白的表达。结果经无氨基酸培养液EBSS处理后48h，HCCLM3、MHCC97L及SMMC-7721细胞存活率分别为（78．9±3．8）％、（57．2±13．1）％及（3．4±3．0）％（P〈0．01）。与HCCLM3、MHCC97L相比，细胞中的自噬体在SMMC-7721细胞中明显增多，分别为（13．1±3．5）％、（20．0±1．1）％及（48．9±4．5）％（P〈0．01）。Western印迹显示在EBSS处理后早期即有LC3—Ⅱ蛋白的明显表达。在EBSS基础上雷帕霉素20μmol／L处理24h，3种细胞株活力明显下降。3-MA、Wortmannin及LY294002处理也加速EBSS诱导的细胞死亡。结论高转移潜能肝癌细胞比较耐受自噬样细胞死亡。P13K／AKT／mTOR信号通路对自噬样细胞死亡可能起重要调节作用。     

磷脂酰肌醇3-激酶信号转导通路在胃肠道间质瘤中的表达及其意义

目的 研究磷脂酰肌醇3-激酶(PI3K/Akt)信号转导通路在胃肠道间质瘤(GIST)中的表达及其意义.方法 用RT-PCR检测PI3K、Akt和PTEN在124例GIST及瘤旁正常组织中的mRNA水平,Western印迹法检测PI3K、Akt和PTEN蛋白在GIST中的表达.结果在GIST中,随着NIH分级增高,PI3K和Akt的mRNA表达水平上调,低危组分别为0.90、0.53,中危组分别为0.88、0.58,高危组分别为0.97、0.58(P<0.05或P<0.01);而抑癌基因PTEN的mRNA表达水平下调,低危组为0.44,中危组为0.37,高危组为0.32(P<0.05或P<0.01).三者阳性表达率与性别、年龄、组织学类型无关,与肿瘤侵袭转移和黏膜受侵关系密切.PI3K与Akt蛋白阳性表达之间呈显著正相关(r=0.292,P:0.031);PI3K与PTEN蛋白阳性表达之间呈显著负相关(r=-0.412,P=0.036);PTEN与Akt蛋白阳性表达之间呈显著负相关(r=-0.386,P=0.024).结论 PI3K/Akt与PTEN在GIST的发生发展中可能互相拮抗、共同作用,可能是恶性肿瘤生长、侵袭、转移的重要指标.     

靶向抑制PI3K对人体外周血破骨细胞分化p38/c-Fos 信号通路调控的研究

目的探讨靶向抑制磷脂酰肌醇3-激酶(PI3K)调控p38/c-Fos信号通路对破骨细胞分化的影响。方法采用人体外周血分离单个核细胞,并诱导形成破骨细胞,干预组应用靶向抑制PI3K的LY294002对细胞进行处理,通过细胞形态学观察并检测细胞活性,再分别采用RT-PCR和Western-Blot两种方法考察p38MAPK通路及下游转录因子c-fos基因和蛋白的表达水平。结果模型组与干预组细胞形态学观察均可见诱导培养的破骨细胞形态明显;与模型组比较,干预组破骨细胞样细胞的形成减少,活性减弱,具有统计学意义(P0.05);RT-PCR结果均显示,干预组p38MAPK通路下游转录因子c-fos表达水平较对照组下降,有显著性差异(P0.01);Western-Blot检测结果显示干预组c-fos蛋白表达水平较对照组降低,具有统计学意义(P0.05)。结论靶向抑制PI3K在体外细胞培养中可抑制破骨细胞形态,并可通过下调p38MAPK通路中c-fos基因和蛋白水平的表达,从而减弱破骨细胞吸收功能。     

PTEN、P13 K和Akt蛋白在膀胱移行细胞癌中的表达及临床意义

目的:探讨PTEN、PI3K和Akt在膀胱移行细胞癌(BTCC)中的表达及其与BTCC临床病理特征的关系,以及PTEN、PI3K和Akt三者之间的相关性。方法:采用免疫组织化学S-P法检测60例BTCC组织和10例正常膀胱黏膜中PTEN、PI3K和Akt的蛋白的表达。结果:BTCC组织中PTEN、PI3K、Akt蛋白的阳性表达率分别为58.33%(35/60)、71.67%(43/60)和56.67%(34/60)。BTCC中PTEN、PI3K、Akt蛋白的阳性表达率与正常膀胱黏膜组织相比,差异均有统计学意义(P<0.05)。在60例BTCC组织中,PTEN分别与PI3K、Akt表达之间均呈显著负相关性(P<0.05),PI3K与Akt表达呈显著正相关性(P=0.001)。结论:PTEN、PI3K、Akt蛋白的表达均与BTCC发生发展及侵袭有关,但PTEN与BTCC发生发展的关系更加密切。在BTCC的发生发展中,PI3K、PTEN的作用可能互相拮抗;PTEN蛋白的表达缺失可能与Akt过表达有关;PI3K和Akt协同促进BTCC的恶性进展。     

Fractalkine deficiency attenuates LPS-induced acute kidney injury and podocyte apoptosis by targeting the PI3K/Akt signal pathway

Clinical and Experimental Nephrology - Podocyte injury is a major biomarker of primary glomerular disease, which leads to massive proteinuria and kidney failure. The increased production of the...     

Targeted RNA interference of PI3K pathway components sensitizes colon cancer cells to TNF-related apoptosis-inducing ligand (TRAIL)

BACKGROUND: The phosphoinositide 3-kinase (PI3K/Akt) pathway transduces signals initiated from growth factors. Previously, we identified an important role for PI3K/Akt in colon cancer progression. The purpose of this study was to determine (1) whether short interfering RNA (siRNA) directed to PI3K/Akt components can render colon cancer cells sensitive to treatment with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and (2) the cellular mechanisms contributing to the enhanced sensitivity. METHODS: Human colon cancer cells KM20 and KM12C (both TRAIL resistant) were transfected with siRNA directed against the PI3K p85alpha regulatory subunit Akt1 or nontargeting control sequence and then treated with TRAIL (100 ng/mL) or vehicle. A ribonuclease protection assay was performed to assess changes in TRAIL receptor expression. Protein was extracted and analyzed by Western blot for expression of cleavage of TRAIL receptors (death receptor (DR) 4 and 5), caspase-3, caspase-8, and BID. Apoptosis was measured by enzyme-linked immunosorbent assay of DNA fragmentation. RESULTS: Combination treatment with p85alpha or Akt1 siRNA and TRAIL increased apoptosis in KM20 and KM12C cells, compared with TRAIL alone; these results were corroborated further by complete inhibition of apoptosis by Z-acetyl-Asp-Glu-Val-Asp-(DEVD)-fmk, a caspase-3 inhibitor. Furthermore, siRNA-mediated PI3K pathway inhibition resulted in increased expression of the TRAIL death receptors 4 and 5. CONCLUSIONS: Inhibition of PI3K/Akt by RNA interference sensitizes resistant colon cancer cells to TRAIL-induced cell death through the induction of TRAIL receptors and activation of caspase-3 and caspase-8. Agents that selectively target the PI3K/Akt pathway may enhance the effects of chemotherapeutic agents and provide novel adjuvant treatment for selected colon cancers.     

Targeted biotherapy in metastatic colorectal carcinoma: Current practice

Targeted therapy has become an indispensable tool in the management of metastatic colorectal cancer (mCRC). The combination of monoclonal antibodies with conventional polychemotherapy has proven its efficacy as the median overall survival now exceeds 24 months: these novel molecules act by targeting circulating vascular endothelial growth factor (VEGF) and the receptor of epidermal growth factor (EGFR). At the present time, no factor has been identified to predict the efficacy of bevacizumab, an inhibitor of circulating VEGF. On the other hand, mutation of the KRAS oncogen has been proven to be a factor of non-response, or even of deleterious response to the use of EGFR, therefore limiting its use to patients whose tumors bear the wild type KRAS oncogen. Treatment toxicity for these molecules is moderate, specific, and is not cumulative with chemotherapy-related toxicity. On the other hand, combined targeted therapy (association of several targeted therapy drugs) has not been shown to be of any benefit. Other biotherapies continue to be developed, but there is not yet a consensus of how to best target the tumor nor which anti-tumoral molecules to use in the treatment of mCRC.     

miR-155靶向调节PIK3R1对类风湿关节炎大鼠模型PI3K/Akt/mTOR信号通路的影响

目的 探讨miR-155靶向调节PIK3R1对类风湿关节炎（rheumatoid arthritis,RA）大鼠PI3K/Akt/mTOR信号通路的影响。方法 SD大鼠随机分为对照组、模型组、miR-155 agomir组、miR-155 antagomir组、miR-155阴性对照组,诱导RA模型,分组处理后,观察关节症状,检测关节炎指数及后足趾容积;HE染色观察大鼠关节组织病理形态;使用试剂盒检测大鼠关节组织炎性因子IL-6、IL-17及IL-18水平;免疫印迹检测大鼠关节组织PI3K/Akt/mTOR信号通路蛋白表达;qRT-PCR实验检测大鼠关节组织miR-155及PIK3R1 mRNA水平;双荧光素酶报告基因实验检测miR-155对PIK3R1的靶向调控作用。结果 与对照组比较,模型组大鼠关节炎症状明显,关节炎指数、后足趾容积、关节组织炎性因子IL-6、IL-17及IL-18水平、关节组织p-PI3K/PI3K、p-Akt/Akt及p-mTOR/mTOR水平、关节组织miR-155水平明显升高（P＜0.05）,PIK3R1 mRNA水平明显降低（P＜0.05）。与模型组、miR-155阴性对照组比较,miR-155 antagomir组大鼠上述指标得到明显改善（P＜0.05）;miR-155 agomir组与miR-155 antagomir组趋势相反（P＜0.05）。结论 miR-155可靶向下调PIK3R1的表达,激活PI3K/Akt/mTOR信号,加重RA大鼠关节炎症损伤,下调miR-155表达,可抑制PI3K/Akt/mTOR信号激活及炎症反应发生发展,改善关节炎症状。     

Targeted therapy in renal cell carcinoma: moving from molecular agents to specific immunotherapy

Non-specific immunotherapy has been for a long time a standard treatment option for patients with metastatic renal cell carcinoma but was redeemed by specific targeted molecular therapies, namely the VEGF and mTOR inhibitors. After moving treatment for mRCC to specific molecular agents with a well-defined mode of action, immunotherapy still needs this further development to increase its accuracy. Nowadays, an evolution from a rather non-specific cytokine treatment to sophisticated targeted approaches in specific immunotherapy led to a re-launch of immunotherapy in clinical studies. Recent steps in the development of immunotherapy strategies are discussed in this review with a special focus on peptide vaccination which aims at a tumor targeting by specific T lymphocytes. In addition, different combinatory strategies with immunomodulating agents like cyclophosphamide or sunitinib are outlined, and the effects of immune checkpoint modulators as anti-CTLA-4 or PD-1 antibodies are discussed.     

磷脂酰肌醇3磷酸激酶信号通路在前列腺癌病程演进中的研究进展

去势和抗雄激素疗法是目前处理转移性前列腺癌的基本手段,但是最终大多数患者演变为雄激素非依赖型癌症而复发,此时的前列腺癌细胞在极低水平的雄激素条件下得以继续生存和恶化。最新研究报告指出磷脂酰肌醇3磷酸激酶（PI3K）信号传导通路的活性升高是前列腺癌细胞在去势条件下得以存活和增殖的关键因素。PI3K信号传导途径还参与雄激素受体（AR）介导的生物学效应,调节前列腺癌细胞的存活与增殖。本文简要阐述PI3K在前列腺癌发展中的关键作用以及与其他信号传导途径的相互调节。     

PI3K突变与出现镶嵌血管的结直肠癌远处转移

目的 探讨出现镶嵌血管的结直肠癌中癌细胞PI3K的突变与结直肠癌远处转移间的关系.方法 收集手术切除,并经免疫组织化学染色证实存在镶嵌血管的168例结直肠癌标本;PCR测序鉴别PI3K突变,并分析其临床、病理与随访资料,进而探讨PI3K突变在出现镶嵌血管的结直肠癌远处转移中的意义.结果 168例出现镶嵌血管的结直肠癌标本,其PI3K突变率为25.0％,外显子9突变率为15.5％,外显子20突变率为9.5％.56例患者术前即出现远处转移,其中发生PI3K突变的为20例,其远处转移率为47.6％,而未出现PI3K突变的36例,其远处转移率为28.6％,两者差异有统计学意义.随访中,发生PI3K突变的复发率及复发伴转移率与未发生PI3K突变相比,其差异均有统计学意义.结论 PI3K突变为出现镶嵌血管结直肠癌转移的相关影响因子之一,在出现镶嵌血管的结直肠癌中检测PI3K基因的突变有利于进一步判断患者预后.