An orally available small-molecule inhibitor of c-Met, PF-2341066, exhibits cytoreductive antitumor efficacy through antiproliferative and antiangiogenic mechanisms

The c-Met receptor tyrosine kinase and its ligand, hepatocyte growth factor (HGF), have been implicated in the progression of several human cancers and are attractive therapeutic targets. PF-2341066 was identified as a potent, orally bioavailable, ATP-competitive small-molecule inhibitor of the catalytic activity of c-Met kinase. PF-2341066 was selective for c-Met (and anaplastic lymphoma kinase) compared with a panel of >120 diverse tyrosine and serine-threonine kinases. PF-2341066 potently inhibited c-Met phosphorylation and c-Met-dependent proliferation, migration, or invasion of human tumor cells in vitro (IC(50) values, 5-20 nmol/L). In addition, PF-2341066 potently inhibited HGF-stimulated endothelial cell survival or invasion and serum-stimulated tubulogenesis in vitro, suggesting that this agent also exhibits antiangiogenic properties. PF-2341066 showed efficacy at well-tolerated doses, including marked cytoreductive antitumor activity, in several tumor models that expressed activated c-Met. The antitumor efficacy of PF-2341066 was dose dependent and showed a strong correlation to inhibition of c-Met phosphorylation in vivo. Near-maximal inhibition of c-Met activity for the full dosing interval was necessary to maximize the efficacy of PF-2341066. Additional mechanism-of-action studies showed dose-dependent inhibition of c-Met-dependent signal transduction, tumor cell proliferation (Ki67), induction of apoptosis (caspase-3), and reduction of microvessel density (CD31). These results indicated that the antitumor activity of PF-2341066 may be mediated by direct effects on tumor cell growth or survival as well as antiangiogenic mechanisms. Collectively, these results show the therapeutic potential of targeting c-Met with selective small-molecule inhibitors for the treatment of human cancers.     

Ras Farnesylation Inhibitor FTI-277 Restores the E-Cadherin/Catenin Cell Adhesion System in Human Cancer Cells and Reduces Cancer Metastasis

The E-cadherin/catenin cell adhesion system is often down-regulated in epithelial tumors. This is thought to play an important role in cancer invasion and metastasis, and restoration of this system may suppress metastatic spread of cancer. In this study, the effects of a Ras farnesylation inhibitor (FTI-277) on E-cadherin-mediated cell-cell adhesion and metastatic potential were examined. In cell aggregation assays, FTI-277 stimulated aggregation of colon, liver and breast cancer cells. In vitro cultures of cancer cells showed that FIT-277 induced strong cell-cell contact. Immunoblotting analysis showed that FTI-277 increased E-cadherin/catenin (α, β and γ) expression and strongly stabilized E-cadherin/catenin with the actin cytoskeleton. Northern blotting studies indicated that the observed increase in the E-cadherin/catenin protein content was due to increased expression of their genes. After inoculation of the spleens of mice with severe combined immunodeficiency (SCID) with cancer cells, FTI-277 treatment for 3 weeks markedly reduced splenic primary tumor growth and the rate of liver metastasis compared with control counterparts. Our data demonstrate that FTI-277 can activate functioning of the E-cadherin-mediated cell adhesion system, which is associated with suppression of cancer cell metastasis. Therefore, selective inhibition of Ras activation may be useful for preventing cancer metastasis.     

ALDH5A1在卵巢癌中的表达及其对细胞侵袭特性和预后的影响

目的探讨醛脱氢酶5A1(ALDH5A1)在卵巢癌组织中的表达、临床意义及其影响卵巢癌细胞侵袭转移的作用和分子机制。方法首先基于GEO数据库比较ALDH5A1在卵巢癌转移组织和原发灶的表达差异。然后通过转染小干扰RNA(siRNA)方式建立ALDH5A1表达下调的卵巢癌细胞株SKOV3,划痕实验和Transwell侵袭实验检测细胞迁移侵袭能力的变化。cBioPortal数据库描绘了ALDH5A1在卵巢癌细胞株和卵巢癌患者中的共表达基因谱。最后运用TCGA和GEO数据库分析ALDH5A1表达水平与卵巢癌预后的相关性,HPA数据库再次确认卵巢癌患者中ALDH5A1和MMPs的相对表达水平。结果相比于卵巢癌原发灶,转移组织中的ALDH5A1表达下调。细胞实验结果显示下调ALDH5A1表达可促进卵巢癌细胞株迁移和侵袭。富集分析发现ALDH5A1的共表达基因在细胞外基质(ECM)组织通路中显著富集,进一步通过数据库转录组数据验证ECM组织通路中发挥重要作用的基质金属蛋白酶(MMP)表达与ALDH5A1表达水平呈负相关,提示ALDH5A1可能通过ECM组织通路影响卵巢癌的转移和侵袭能力。生存分析结果提示ALDH5A1低表达的卵巢癌患者预后更差。结论ALDH5A1表达下调可能促进卵巢癌侵袭转移,且与预后不良相关。     

Recombinant Human Endostatin Endostar Inhibits Tumor Growth and Metastasis in a Mouse Xenograft Model of Colon Cancer

To investigate the effects of recombinant human endostatin Endostar on metastasis and angiogenesis and lymphangiogenesis of colorectal cancer cells in a mouse xenograft model. Colon cancer cells SW620 were injected subcutaneously into the left hind flank of nude mice to establish mouse xenograft models. The mice were treated with normal saline or Endostar subcutaneously every other day. The growth and lymph node metastasis of tumor cells, angiogenesis and lymphangiogenesis in tumor tissue were detected. Apoptosis and cell cycle distribution were studied by flow cytometry. The expression of VEGF-A, -C, or -D in SW620 cells was determined by immunoblotting assays. Endostar inhibited tumor growth and the rate of lymph node metastasis (P < 0.01). The density of blood vessels in or around the tumor area was 12.27 ± 1.21 and 22.25 ± 2.69 per field in Endostar-treated mice and controls (P < 0.05), respectively. Endostar also decreased the density of lymphatic vessels in tumor tissues (7.84 ± 0.81 vs. 13.83 ± 1.08, P < 0.05). Endostar suppresses angiogenesis and lymphangiogenesis in the lymph nodes with metastases, simultaneously. The expression of VEGF-A, -C and -D in SW620 cells treated with Endostar was substantially lower than that of controls. Endostar inhibited growth and lymph node metastasis of colon cancer cells by inhibiting angiogenesis and lymphangiogenesis in a mouse xenograft model of colon cancer.     

c-Met 、TIMP2 与宫颈鳞癌浸润转移的相关性

 目的 研究肝细胞生长因子的受体c-Met与基质金属蛋白酶抑制剂TIMP2在宫颈鳞癌组织中表达的意义及其与宫颈鳞癌临床分期、病理分级和淋巴结转移的关系。方法 采用SP法检测40例宫颈鳞癌、13例宫颈上皮内瘤样病变（CIN）、15例正常宫颈组织中c-Met和TIMP2的表达，对结果进行秩和检验分析。结果 正常宫颈、CIN和宫颈鳞癌组织中c-Met表达依次增高，宫颈癌的临床分期越晚，c-Met表达越强。TIMP2表达CIN中明显高于正常组织，到宫颈鳞癌又呈下降趋势，临床分期越晚，表达越弱。有淋巴结转移组与无淋巴结转移组比较，c-Met表达增高，TIMP2表达降低，而两者均与病理分级无关。结论 c-Met与宫颈鳞癌发生发展有关，TIMP2可能是宫颈鳞癌发展过程中的一早期事件。c-Met、TIMP2可成为判断宫颈癌转移、预后的良好参考指标。     

卵巢癌肠道转移的危险因素及预后分析

[目的]分析卵巢癌肠道转移的危险因素。[方法]回顾性分析2005年6月至2008年6月经手术治疗的235例晚期卵巢恶性肿瘤病例的临床资料,经术后病理证实肠道转移者107例。[结果]吸烟、体重指数〉30kg/m2、肿瘤直径〉4cm、原发肿瘤累及两侧卵巢、临床分期较晚、组织分化较差、伴有大量腹水（≥1500ml）、CA125较高（≥500U/ml）与卵巢癌肠道转移有关（P〈0.05）。转移肠段切除患者3年和5年生存率分别为57.7%和43.8%,肿瘤局部切除患者3年和5年生存率分别为30.4%和12.2%,两组3年和5年生存率差异有统计学意义（P〈0.05）。残余病灶≥2cm的患者半年内复发率（54.5%）比残余病灶〈2cm患者（18.8%）高（P〈0.05）,首次切除的患者半年内复发率（24.4%）比两次或以上切除患者（55.2%）低（P〈0.05）。[结论]为提高卵巢癌肠道转移者疗效,术前应全面分析肠道转移的危险因素,评估肠道受累程度,做好充分的肠道准备;术中行肿瘤细胞减灭术应尽量采取转移肠道切除术。     

卵巢癌细胞上皮间质转化及侵袭转移过程中microRNA-9的调控作用

目的 探讨microRNA-9（miR-9）参与调控卵巢癌细胞EMT过程,以及影响卵巢癌细胞侵袭及转移的分子机制。方法 使用TargetScan及PicTar数据库,预测可能靶向E-cadherin 3’UTR区的miRNA,双荧光素酶报告体系进行验证;上调候选miR后,用qRT-PCR和Western blot检测E-cadherin的表达变化;细胞免疫荧光观察E-cadherin、β-Catenin和Vimentin的表达;划痕实验、Transwell实验,观察卵巢癌细胞运动和侵袭能力的改变。结果 TargetScan、PicTar预测发现miR-9是唯一可与CDH1结合的miRNA。双荧光素酶报告系统验证预测结果正确。在SKOV-3中上调miR-9后,E-cadherin的表达受到显著抑制;细胞形态向间质细胞样转变,发生EMT分子水平的特征性改变;体外实验表明,卵巢癌细胞的运动和侵袭能力得到明显促进。结论 miR-9可以通过靶向调控E-cadherin表达,促进卵巢癌细胞的EMT进程,对卵巢癌细胞的运动和侵袭能力产生重要影响。     

nm23及NDRG1 蛋白在卵巢癌肿瘤抑郁模型中的表达*

目的:探讨不良心理应激影响肿瘤发生发展的分子学机制。方法:采用蛋白组学方法筛选、鉴定荷卵巢癌裸鼠不良心理应激模型移植瘤中的差异表达蛋白,并用Western Blot验证;用免疫组化SP法检测25例应激组和20例对照组移植瘤中的nm23及NDRG 1 蛋白的表达特点。结果:两组在pI 7.1、Mr=21ku和pI 5.5、Mr=43ku处有两个显著差异点,前一个在应激组表达上调而后一个表达下调,通过质谱分别鉴定为nm23及NDRG 1 蛋白;Western Blot验证结果同蛋白组学的一致;免疫组化显示nm23蛋白在应激组和对照组中阳性表达率分别为100% 和70% ,NDRG 1 蛋白为56% 和85% ,差异均有统计学意义（P<0.05）。 结论:不良心理应激可能通过激活癌基因和失活抑癌基因来影响肿瘤的发生发展,nm23及NDRG 1 可作为基因研究和基因治疗的靶点。      

结肠癌中肿瘤相关巨噬细胞与肿瘤浸润转移的关系

[目的]了解结肠癌组织中肿瘤相关巨噬细胞的浸润情况,探讨其与结肠癌侵袭转移的相关性。[方法]采用免疫组化染色的方法,对69例结肠癌标本中巨噬细胞的浸润情况进行检测,同时行巨噬细胞、淋巴管计数,并对巨噬细胞数量与临床病理参数之间的关系进行统计分析。[结果]结肠癌组织中有大量的肿瘤相关巨噬细胞浸润。结肠癌巨噬细胞计数与Duke’s分期有相关性(P=0.023),分期偏晚者巨噬细胞计数高于分期偏早者。巨噬细胞计数在年龄、性别、肿瘤分化程度等方面不存在差异(P>0.05)。巨噬细胞数量也与淋巴管数量相关(r=0.432,P<0.01)。[结论]结肠癌肿瘤组织中肿瘤相关巨噬细胞与淋巴管生成及肿瘤的侵袭转移有关。     

Progesterone and Estradiol Synergistically Promote the Lung Metastasis of Tuberin-Deficient Cells in a Preclinical Model of Lymphangioleiomyomatosis

Lymphangioleiomyomatosis (LAM) is a female-predominant lung disease that can lead to respiratory failure. LAM cells typically have inactivating tuberous sclerosis 2 (TSC2) mutations, leading to mTORC1 hyperactivation. The gender specificity of LAM suggests that female hormones contribute to disease progression. Clinical findings indicate that estradiol exacerbates LAM behaviors and symptoms. Although hormonal therapy with progesterone has been employed, the benefit in LAM improvement has not been achieved. We have previously found that estradiol promotes the survival and lung metastasis of cells lacking tuberin in a preclinical model of LAM. In this study, we hypothesize that progesterone alone or in combination with estradiol promotes metastatic behaviors of TSC2-deficient cells. In cell culture models of TSC2-deficient LAM patient-derived and rat uterine leiomyoma-derived cells, we found that progesterone treatment or progesterone plus estradiol resulted in increased phosphorylation of Protein Kinase B (Akt) and Extracellular signal-regulated kinases1/2 (ERK1/2), induced the proliferation, and enhanced the migration and invasiveness. In addition, treatment of progesterone plus estradiol synergistically decreased the levels of reactive oxygen species and enhanced cell survival under oxidative stress. In a murine model of LAM, treatment of progesterone plus estradiol promoted the growth of xenograft tumors; however, progesterone treatment did not affect the development of xenograft tumors of Tsc2-deficient cells. Importantly, treatment of progesterone plus estradiol resulted in alteration of lung morphology and significantly increased the number of lung micrometastases of Tsc2-deficient cells compared with estradiol treatment alone. Collectively, these data indicate that progesterone increases the metastatic potential of Tsc2-deficient LAM patient-derived cells in vitro and lung metastasis in vivo. Thus, targeting progesterone-mediated signaling events may have therapeutic benefit for LAM and possibly other hormonally dependent cancers.     

肿瘤相关巨噬细胞与胃癌血管生成及转移关系

[目的]探讨肿瘤相关巨噬细胞（TAMs）浸润与胃癌血管生成及转移的关系。[方法]采用免疫组织化学法检测51例胃癌组织微血管密度（MVD）、TAMs和血管内皮生长因子（VEGF）的表达。[结果]TAMs的表达在高分化癌组织中明显低于中-低分化组（P〈0．05）；淋巴结转移组高于无淋巴结转移组（P〈0．05）；TAMs与VEGF及MVD均呈正相关（r=0．74，P〈0．01：r=0．58，P〈0．05）。[结论]胃癌组织中TAMs在血管生成及肿瘤转移中可能起重要作用。     

人卵巢上皮癌裸鼠皮下移植瘤模型的建立及生物学性状的鉴定

目的　建立人卵巢上皮性癌裸鼠皮下移植瘤模型 ,为人卵巢癌的诊断及治疗诸方面提供理想的在体研究工具。方法体外培养人卵巢上皮癌细胞株SKOV 3 ,采用皮下注射方式接种至裸小鼠 ,待肿瘤直径达 0 .8cm～ 1.0cm时 ,将其切下剪成直径为1mm～ 2mm小块 ,再接种至另一裸鼠皮下进行传代。病理组织学检查或透射电镜观察肿瘤 ,并观察染色体特征 ,检测裸鼠血清和肿瘤组织中肿瘤标志物CA12 5、CEA、AFP的浓度。结果　成功地建立了人卵巢上皮性癌细胞株SKOV 3裸小鼠皮下移植瘤模型 ,并传至第二代 ,原代接种成功率为 69.2 % (2 7/ 3 9) ,鼠间传代移植成功率为 10 0 .0 % ,具备人肿瘤生物学特点 ,但未观察到转移行为。裸鼠血清CA 12 5浓度低 ,但在肿瘤冻融产物中较高。结论　建立的人卵巢癌裸鼠皮下移植瘤模型具有人卵巢癌特征 ,可用于研究卵巢癌的生物学特性 ,为卵巢癌临床诊断及治疗研究提供有用工具。     

1990—2019年中国卵巢癌疾病负担及其变化趋势分析

[目的]分析1990—2019年中国卵巢癌疾病负担长期变化趋势。[方法]通过全球疾病负担研究（GBD）数据,应用Joinpoint 4.9.1.0软件分析1990—2019年中国卵巢癌标化患病率、发病率、死亡率、伤残调整寿命年（DALYs）、过早死亡损失寿命年（YLLs）和伤残损失寿命年（YLDs）等指标的平均年度变化百分比（AAPC）。[结果] 1990—2019年中国卵巢癌的患病数、发病数和死亡数均有明显增加。标化患病率从10.11/10万增加至20.44/10万,标化发病率从2.56/10万增加至4.54/10万,标化死亡率从1.76/10万增加至2.77/10万,标化DALYs率从55.57/10万增加至80.52/10万。标化患病率、标化发病率、标化死亡率、标化DALYs率、标化YLLs率和标化YLDs率整体均呈上升趋势,AAPC值分别为2.48%、2.02%、1.58%、1.32%、1.32%和2.26%,差异均具有统计学意义（P<0.001）。从35岁开始卵巢癌疾病负担快速上升,发病率和DALYs率在65～69岁组最高,死亡率在≥70岁组最高。[结论] 1990—2...     

Fas/FasL系统与卵巢肿瘤的免疫逃逸的研究

目的 :检测Fas、FasL在上皮性卵巢癌及其肿瘤浸润淋巴细胞 (TIL)中的表达 ,探讨Fas系统在卵巢癌免疫逃逸中的作用。方法 :用流式细胞术检测了 31份上皮性卵巢癌、2 0份卵巢良性肿瘤、10份正常卵巢组织以及 31份卵巢癌TIL、12份卵巢良性肿瘤TIL的Fas及FasL的表达。结果 :卵巢癌组织的Fas表达明显低于卵巢良性肿瘤及正常卵巢组织 ,P <0 0 1;而其FasL表达明显高于后者 ,P <0 0 1。卵巢良性肿瘤及正常卵巢组织的Fas、FasL表达差异无显著意义 ,P >0 0 5。卵巢癌组织中含有丰富的TIL ,其Fas及FasL的表达明显高于卵巢良性肿瘤TIL ,差异有显著意义 ,P <0 0 5。卵巢癌组织Fas的表达与临床分期、组织学分级及淋巴结转移无关 ,P >0 0 5。FasL的表达与临床分期无关 ,但随组织学分级的升高而增加 ,P <0 0 5 ,有淋巴结转移者FasL的表达明显高于无淋巴结转移者 ,P <0 0 5。卵巢癌TIL中Fas及FasL的表达与各临床病理参数无关 ,P >0 0 5。结论 :上皮性卵巢癌组织中存在Fas表达的下调和FasL表达的增加 ,FasL高表达者预后不良。肿瘤细胞可能通过FasL的过度表达 ,逃避免疫监视 ,诱导Fas敏感的TIL凋亡 ,发生浸润和转移。     

Effects of Valproic Acid on Proliferation,Apoptosis, Angiogenesis and Metastasis of Ovarian Cancer in Vitro and in Vivo

Inhibitors of histone deacetylase activity are emerging as a potentially important new class of anticanceragents. In this study, we assessed the anticancer effects of valproic acid (VPA) on ovarian cancer in vitro and invivo. Cultured SKOV3 cells were treated by VPA with different concentrations and time, then the effects on cellgrowth, cell cycle, apoptosis, and related events were investigated. A human ovarian cancer model transplantedsubcutaneously in nude mice was established, and the efficacy of VPA used alone and in combination withdiammine dichloroplatinum (DDP) to inhibit the growth of tumors was also assessed. Proliferation of SKOV3 cellswas inhibited by VPA in a dose and time dependent fashion. The cell cycle distribution changed one treatmentwith VPA, with decrease in the number of S-phase cells and increase in G1-phase. VPA could significantly inhibitthe growth of the epithelial ovarian cancer SKOV3 cells in vivo without toxic side effects. Treatment with VPAcombined with DDP demonstrated enhanced anticancer effects. The result of flow cytometry (FCM) indicatedthat after VPA in vitro and in vivo, the expression of E-cadherin was increased whereas vascular endothelialgrowth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were decreased. This study suggests that VPAcould be a novel attractive agent for treatment of ovarian cancer.     

程序性细胞死亡6对促进卵巢癌细胞增殖和转移作用的效应分子研究

[目的]PDCD6是一种程序性细胞死亡基因。近年来,已有不少研究证明PDCD6在一定程度上可以促进卵巢癌细胞的增殖和迁移。然而,它的确切分子机制还有待进一步验证。[方法]利用慢病毒sh RNA敲除HO-8910PM的PDCD6水平,以空载体作为阴性对照。提取两组细胞的总RNA,用于Allumila基因芯片分析,每组实验重复2次,并用荧光定量PCR验证表达有差异的基因。[结果]基因芯片结果表明,PDCD6敲除后共有133个基因表达有显著差异。其中,101个基因表达下调,32个基因表达上调。基因功能分析显示它们分别与细胞凋亡、增殖、周期、迁移和血管生成等有关。信号通路富集度显示MAPK信号通路激活,主要激活的基因有FGFR、MAP2K1/MEK1、MAP2K2/MEK2、MYC、FOS。RT-PCR验证结果显示,PDCD6敲除后CCND1、MYC、ANGPTL4、BMP2、CXCL16基因m RNA的表达比对照组明显下调,与基因芯片的结果相同。[结论]PDCD6可能通过上调CCND1、MYC、ANGPT4、BMP2和CXCL16基因,激活MAPK信号通路,从而促进卵巢癌细胞的增殖和转移。