Scoparone inhibits pancreatic cancer through PI3K/Akt signaling pathway

BACKGROUND Pancreatic cancer is a highly malignant tumor of the gastrointestinal system whose emerging resistance to chemotherapy has necessitated the development of novel antitumor treatments.Scoparone,a traditional Chinese medicine monomer with a wide range of pharmacological properties,has attracted considerable attention for its antitumor activity.AIM To explore the potential antitumor effect of scoparone on pancreatic cancer and the possible molecular mechanism of action.METHODS The target genes of scoparone were determined using both the bioinformatics and multiplatform analyses.The effect of scoparone on pancreatic cancer cell proliferation,migration,invasion,cell cycle,and apoptosis was detected in vitro.The expression of hub genes was tested using quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the molecular mechanism was analyzed using Western blot.The in vivo effect of scoparone on pancreatic cancer cell proliferation was detected using a xenograft tumor model in nude mice as well as immunohistochemistry.RESULTS The hub genes involved in the suppression of pancreatic cancer by scoparone were obtained by network bioinformatics analyses using publicly available databases and platforms,including SwissTargetPrediction,STITCH,GeneCards,CTD,STRING,WebGestalt,Cytoscape,and Gepia;AKT1 was confirmed using qRT-PCR to be the hub gene.Cell Counting Kit-8 assay revealed that the viability of Capan-2 and SW1990 cells was significantly reduced by scoparone treatment exhibiting IC50 values of 225.2μmol/L and 209.1μmol/L,respectively.Wound healing and transwell assays showed that scoparone inhibited the migration and invasion of pancreatic cancer cells.Additionally,flow cytometry confirmed that scoparone caused cell cycle arrest and induced apoptosis.Scoparone also increased the expression levels of Bax and cleaved caspase-3,decreased the levels of MMP9 and Bcl-2,and suppressed the phosphorylation of Akt without affecting total PI3K and Akt.Moreover,compared with the control group,xenograft tumors,in the 200μmol/L scoparone treatment group,were smaller in volume and lighter in weight,and the percentages of Ki65-and PCNA-positive cells were decreased.CONCLUSION Our findings indicate that scoparone inhibits pancreatic cancer cell proliferation in vitro and in vivo,inhibits migration and invasion,and induces cycle arrest and apoptosis in vitro through the PI3K/Akt signaling pathway.

Scorpiones,Scolopendra and Gekko Inhibit Lung Cancer Growth and Metastasis by Ameliorating Hypoxic Tumor Microenvironment via PI3K/AKT/mTOR/HIF-1αSignaling Pathway

Objective:To investigate whether Buthus martensii karsch(Scorpiones),Scolopendra subspinipes mutilans L.Koch(Scolopendra)and Gekko gecko Linnaeus(Gekko)could ameliorate the hypoxic tumor microenvironment and inhibit lung cancer growth and metastasis by regulating phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin/hypoxia-inducible factor-1α(PI3K/AKT/mTOR/HIF-1α)signaling pathway.Methods:Male C57BL/6J mice were inoculated with luciferase labeled LL/2-luc-M38 cell suspension to develop lung cancer models,with rapamycin and cyclophosphamide as positive controls.Carboxy methyl cellulose solutions of Scorpiones,Scolopendra and Gekko were administered intragastrically as 0.33,0.33,and 0.83 g/kg,respectively once daily for 21 days.Fluorescent expression were detected every 7 days after inoculation,and tumor growth curves were plotted.Immunohistochemistry was performed to determine CD31 and HIF-1αexpressions in tumor tissue and microvessel density(MVD)was analyzed.Western blot was performed to detect the expression of PI3K/AKT/mTOR/HIF-1αsignaling pathway-related proteins.Enzyme-linked immunosorbent assay was performed to detect serum basic fibroblast growth factor(bFGF),transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF)in mice.Results:Scorpiones,Scolopendra and Gekko prolonged the survival time and inhibited lung cancer metastasis and expression of HIF-1α(all P<0.01).Moreover,Scorpiones,Scolopendra and Gekko inhibited the phosphorylation of AKT and ribosomal protein S6 kinase(p70S6K)(P<0.05 or P<0.01).In addition,they also decreased the expression of CD31,MVD,bFGF,TGF-β1 and VEGF compared with the model group(P<0.05 or P<0.01).Conclusion:Scorpiones,Scolopendra and Gekko all showed beneficial effects on lung cancer by ameliorating the hypoxic tumor microenvironment via PI3K/AKT/mTOR/HIF-1αsignaling pathway.

Ramulus Cinnamomi extract attenuates neuroinflammatory responses via downregulating TLR4/MyD88 signaling pathway in BV2 cells

Ramulus Cinnamomi （RC）, a traditional Chinese herb, has been used to attenuate inflammatory responses. The purpose of this study was to investigate the effect of RC extract on lipopolysaccharide （LPS）-induced neuroinflammation in BV2 microglial cells and the underlying mechanisms involved. BV2 cells were incubated with normal medium （control group）, LPS, LPS plus 30 pg/mL RC extract, or LPS plus 100 pg/mL RC extract. The BV2 cell morphology was observed under an optical microscope and cell viability was detected by MTT assay. Nitric oxide level in BV2 cells was detected using Griess regents, and the levels of interleukin-6, interleukin-1 β, and tumor necrosis factor u in BV2 cells were determined by ELISA. The expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 proteins were detected by western blot assay. Compared with the LPS group, both 30 and 100 μg/mL RC extract had no significant effect on the viability of BV2 cells. The levels of nitric oxide, interleukin-6, interleukin-1β and tumor necrosis factor ct in BV2 cells were all significantly increased after LPS induction, and the levels were significantly reversed after treatment with 30 and 100 μg/mL RC extract. Furthermore, RC extract significantly inhibited the protein expression levels of cyclooxygenase-2, Toll-like receptor 4 and myeloid differentiation factor 88 in LPS-induced BV2 cells. Our findings suggest that RC extract alleviates neuroinflammation by downregulating the TLR4/MyD88 signaling pathway.

Molecular regulation of vasculogenic mimicry in tumors and potential tumor-target therapy

"Vasculogenic mimicry(VM)",is a term that describes the unique ability of highly aggressive tumor cells to express a multipotent,stem cell-like phenotype,and form a pattern of vasculogenic-like networks in threedimensional culture.As an angiogenesis-independent pathway,VM and/or periodic acid-schiff-positive patterns are associated with poor prognosis in tumor patients.Moreover,VM is resistant to angiogenesis inhibitors.Here,we will review the advances in research on biochemical and molecular signaling pathways of VM in tumors and on potential anti-VM therapy strategy.

Transferrin receptor-targeted immunostimulant for photodynamic immunotherapy against metastatic tumors through b-catenin/CREB interruption

The immunosuppressive phenotype of tumor cells extensively attenuates the immune activation effects of traditional treatments.In this work,a transferrin receptor(TfR)targeted immunostimulant(PTI)is fabricated for photodynamic immunotherapy against metastatic tumors by interrupting b-catenin signal pathway.To synthesize PTI,the photosensitizer conjugated TfR targeting peptide moiety(Palmitic-K(PpIX)-HAIYPRH)is unitized to encapsulate the transcription interrupter of ICG-001.On the one hand,the recognition of PTI and TfR can promote drug delivery into tumor cells to destruct primary tumors through photodynamic therapy and initiate an immunogenic cell death with the release of tumorassociated antigens.On the other hand,PTI will interrupt the binding between b-catenin andcAMP response element-binding protein(CREB),regulating the gene transcription to downregulate programmed death ligand 1(PD-L1)while upregulating CeC motif chemokine ligand 4(CCL4).Furthermore,the elevated CCL4 can recruit the dendritic cells to present tumor-specific antigens and promote T cells activation and infiltration,and the downregulated PD-L1 can avoid the immune evasion of tumor cells and activate systemic anti-tumor immunity to eradicate lung metastasis.This work may inspire the development of antibody antibody-free strategy to activate systemic immune response in consideration of immunosuppressive conditions.

PI3K signaling pathway targeting by using different molecular approaches to treat cancer

Recent evidence of research has been proposed that the phosphoinositide 3-kinase(PI3K) pathway is noticeable target for searching novel anticancer agents. The phosphoinositide 3-kinase(PI3K) is accountable for harmonizing a diverse range of cell functions, such as transcription, proliferation, cell survival, cell growth, degranulation, vesicular trafficking and cell migration, which are mostly involved in carcinogenesis. Particularly, PI3K-mediated signaling molecules and its effects on gene expression contribute to tumorigenesis. PI3Ks generally are grouped into three distinct classes: Ⅰ, Ⅱ and Ⅲ according to their structure and function. The class IA of PI3K includes an alpha, beta or delta p110 catalytic subunit(p110α, p110β, or p110γ), which are associated with the activation of RTKs. Mutations in PIK3CA, the gene encoding the p110α catalytic subunit of PI3K, have just been recognized as novel mechanisms of inducing oncogenic PI3K signaling. Therefore, the class IA PI3K is the only one of most evidently implicated in cancer. The PI3K pathway is mostly mutated in more cancer patients compared with normal person, making it an eyecatching molecular target for analyses based on inhibitor molecule. In this article, we highlighted the signaling effects and regulation pathway of PI3K involved in the development and survival of tumor cells. The consequence and intricacy of PI3K pathway made it an essential beneficial target for cancer treatment.

Emerging role of Hippo pathway in gastric and othergastrointestinal cancers

More evidence has underscored the importance of Hippo signaling pathway in gastrointestinal tissue homeostasis, whereas its deregulation induces tumorigenesis. Yes-associated protein 1(YAP1) and its close paralog TAZ, transcriptional co-activator with a PDZbinding motif, function as key effectors negatively controlled by the Hippo pathway. YAP1/TAZ exerts oncogenic activities by transcriptional regulation via physical interaction with TEAD transcription factors. In various cancers, Hippo pathway cross-talks with pro- or anti-tumorigenic pathways such as GPCR, Wnt/β-catenin, Notch and TGF-β signaling and is deregulated by multiple factors including cell density/junction and micro RNAs. As YAP1 expression is significantly associated with poor prognosis of gastric and other gastrointestinal cancers, detailed delineation of Hippo regulation in tumorigenesis provides novel insight for therapeutic intervention. In current review, we summarized the recent research progresses on the deregulation of Hippo pathway in the gastrointestinal tract including stomach and discuss the molecular consequences leading to tumorigenesis.

Novel molecular targets in hepatocellular carcinoma

Globally,hepatocellular carcinoma(HCC)is a leading cause of cancer and cancerrelated deaths.The therapeutic efficacy of locoregional and systemic treatment in patients with advanced HCC remains low,which results in a poor prognosis.The development of sorafenib for the treatment of HCC has resulted in a new era of molecular targeted therapy for this disease.However,the median overall survival was reported to be barely higher in the sorafenib treatment group than in the control group.Hence,in this review we describe the importance of developing more effective targeted therapies for the management of advanced HCC.Recent investigations of molecular signaling pathways in several cancers have provided some insights into developing molecular therapies that target critical members of these signaling pathways.Proteins involved in the Hedgehog and Notch signaling pathways,Polo-like kinase 1,arginine,histone deacetylases and Glypican-3 can be potential targets in the treatment of HCC.Monotherapy has limited therapeutic efficacy due to the development of inhibitory feedback mechanisms and induction of chemoresistance.Thus,emphasis is now on the development of personalized and combination molecular targeted therapies that can serve as ideal therapeutic strategies for improved management of HCC.

Discoidin domain receptor 1 as a potent therapeutic target in solid tumors

Despite significant discoveries in basic cancer research and improvements in treatment options and clinical outcomes,cancer remains a major public health concern worldwide.Today,the main focus of cancer research is the signaling pathways that are crucial for cell survival,cell proliferation,and cell migration.The aberrant expression of proteins involved in these signaling pathways often leads to abnormal cell growth,cell metastasis,and invasion of healthy tis-sues.One such protein is discoidin domain receptor 1(DDR1)which belongs to the family of receptor tyrosine kinases(RTKs)and is activated upon collagen binding,as a result,downstream signaling pathways are stimulated which are responsible for cell survival,cell growth,adhesion,extracellular matrix remodeling,and cell migration.DDR1 is found to have abnormally elevated expression in various solid tumors,implying a critical role in cancer progression.Tradi-tional cancer treatment involves the use of cytotoxic drugs,chemotherapy,radiotherapy,and surgery,which do not pro-vide long-term survival and often result in cancer recurrence.Numerous small-molecule kinase inhibitors have been synthesized against RTKs including DDR1 and have been highly efficacious in tumor reduction.More recently,targeting the DDR1 extracellular domain(ECD)has garnered much attention from researchers,as inhibiting the DDR1-collagen binding has been attributed to maximizing the likelihood of the combined cytotoxic effect of both immune cells and tar-geted drugs.This review focuses on the structure,function,activation,and signaling partners of DDR1,its role in different solid tumors,andfinally discusses about designing more DDR1 non-kinase inhibitors as promising therapeutic strategies against DDR1-driven tumors.

星形细胞上调基因-1在肿瘤中的研究进展

星形细胞上调基因-1(AEG-1)最早于2002年被发现并成功克隆。AEG-1是一种癌基因,在多种肿瘤中高表达,与肿瘤分化、淋巴结转移、TNM分期和不良预后有关。AEG-1基因能激活PI3K/AKT、NF-κB、MAPK、Wnt等重要信号通路,通过改变下游基因的表达调控细胞的生理和病理过程,包括肿瘤的增殖、侵袭、转移、血管的形成及耐药性等。研究发现,AEG-1对于恶性肿瘤的诊断及预后的判断具有重要的参考价值,其有望成为肿瘤预后评估的重要指标及基因治疗的新靶点。

卵巢癌组织中DNMT1、STAT2表达变化及临床意义

目的:探讨卵巢癌组织中DNA甲基转移酶-1(DNMT1)、信号转导及转录活化因子2(STAT2)表达及临床意义。方法:选取2017年7月-2021年7月本院行手术切除卵巢癌患者80例为卵巢癌组,同期因子宫肌瘤、子宫腺肌症行全子宫+双附件切除术患者80例为对照组,应用免疫组化法测定卵巢癌组织与正常卵巢组织中DNMT1、STAT2表达,分析卵巢癌组织中DNMT1、STAT2表达相关性及与临床病理参数的关系,以Kaplan-Meier生存曲线分析癌组织中DNMT1、STAT2表达与患者1年生存率的关系,Cox分析卵巢癌患者预后影响因素。结果:卵巢癌组DNMT1(83.8%)、STAT2(67.5%)表达阳性率均高于对照组(21.3%、8.8%)(P<0.05);FIGOⅢ+Ⅳ期、肿瘤分化程度低分化、有淋巴结转移、有腹水卵巢癌患者癌组织中DNMT1、STAT2表达阳性率高于Ⅰ+Ⅱ期、中、高分化、无淋巴结转移、无腹水患者(P<0.05);Spearman相关性分析显示,卵巢癌组织中DNMT1、STAT2表达呈显著正相关(P<0.05);KaplanMeier生存曲线分析显示,卵巢癌组DNMT1表达阳性与阴性患者1年总生存率(72.3%、92.3%)无差异(P>0.05),STAT2表达阴性患者1年总生存率(92.0%)高于阳性患者(67.9%)(P<0.05);行Cox分析发现,FIGO分期、肿瘤分化程度、淋巴结转移及癌组织中STAT2高表达均为患者1年预后死亡的影响因素(P<0.05)。结论:卵巢癌组织中DNMT1、STAT2表达显著增高,两者可能协同参与了卵巢癌发生发展及转移,且与患者1年预后有一定相关性。

瘤组织中O-岩藻糖基转移酶1基因表达变化对相关肿瘤预后的影响及其作用机制

目的探讨癌组织中O-岩藻糖基转移酶1基因(POFUT1)的表达变化对相关肿瘤预后的影响及其作用机制。方法结合TCGA数据库及GTEx数据库获取的泛癌数据集,利用GEPIA2及TIMER2数据集,比较多种类型肿瘤与配对的正常组织中POFUT1的表达水平,选出比较有统计学差异的肿瘤类型;比较不同POFUT1水平的上述肿瘤患者的预后。利用TIMER数据库及Estimate软件包筛选POFUT1与免疫细胞浸润水平关联性最强的肿瘤,并分析POFUT1与上述肿瘤组织基质评分、免疫细胞浸润评分及综合评分的相关性。进一步利用String数据库及GE⁃PIA2筛选POFUT1关联基因,并通过DAVID数据库对POFUT1及其关联基因进行信号通路富集分析。结果与配对的正常组织比较,POFUT1在胆管癌、结肠癌、乳腺癌、食管癌、胶质细胞瘤、头颈鳞状细胞癌、肾嫌色细胞癌、脑低级别胶质瘤、肝细胞癌、肺腺癌、肺鳞癌、胰腺癌、直肠腺癌、皮肤黑色素瘤、胃癌、胃食管癌、甲状腺癌组织中的表达差异具有统计学意义(P均<0.05)。POFUT1在TIMER2和GEPIA数据库中同时均表达差异的肿瘤类型有7种(结肠癌、食管癌、胶质细胞瘤、头颈鳞状细胞癌、肾嫌色细胞癌、直肠腺癌、胃癌)。肾透明细胞癌和直肠腺癌中,POFUT1高表达组的总生存期高于POFUT1低表达组(P均<0.05);脑低级别胶质瘤、间皮瘤、葡萄膜黑色素瘤中,POFUT1低表达组总生存期高于POFUT1高表达组(P均<0.05)。POFUT1与免疫细胞浸润水平关联性最强的3种肿瘤分别为脑低级别胶质瘤、结肠癌、膀胱尿路上皮癌;POFUT1表达水平与脑低级别胶质瘤组织的基质、免疫细胞浸润、综合评分及结直肠癌组织的免疫细胞浸润评分均具有相关性(P均<0.05)。POFUT1及其关联基因富集的通路主要为蛋白质结合及RNA结合途径、丝氨酸相关酶途径、细胞凋亡通路和Notch信号通路。结论POFUT1在17种肿瘤中表达水平上升;POFUT1高表达的肾透明细胞癌和直肠腺癌预后好,POFUT1低表达的脑低级别胶质瘤、间皮瘤、葡萄膜黑色素瘤预后好;POFUT1可能通过调控肿瘤免疫浸润水平及信号通路如蛋白质结合及RNA结合途径、Notch信号通路等影响肿瘤患者预后。

WTAPP1对肾母细胞瘤细胞增殖、侵袭、迁移及Wnt/β-catenin信号通路的影响

目的:探讨长链非编码RNA Wilms瘤1相关蛋白假基因1(Wilms tumor 1associated protein pseudogene 1,WTAPP1)对肾母细胞瘤细胞增殖、侵袭、迁移及Wnt/β-catenin信号通路的影响。方法:实时荧光定量PCR法检测48例肾母细胞瘤组织及及其配对的癌旁组织、人肾母细胞瘤细胞(SK-NEP-1)和正常肾上皮细胞(PCS-400-010、PCS-400-011和PCS-400-012)中WTAPP1的相对表达量。分析比较WTAPP1高表达和低表达组患者的临床病理特征及预后。采用慢病毒感染的方法将携带有WTAPP1全基因(过表达WTAPP1)的慢病毒载体或特异性针对WTAPP1基因的shRNA(shWTAPP1)的慢病毒载体分别转入SK-NEP-1细胞。分别采用克隆形成实验和CCK-8法检测SK-NEP-1细胞的增殖能力,细胞划痕愈合实验和Transwell小室实验检测细胞迁移和侵袭能力,蛋白质印迹法检测SK-NEP-1细胞中Wnt3a、β-catenin、C-myc和Survivin蛋白的相对表达量。结果:癌组织中WTAPP1的相对表达量明显高于癌旁组织(P<0.05)。SK-NEP-1细胞WTAPP1的表达水平明显高于正常肾上皮细胞系(PCS-400-010、PCS-400-011和PCS-400-012细胞)(P<0.05)。WTAPP1表达与肾母细胞瘤临床分期具有相关性(P<0.05),而与患者的年龄、性别、肿瘤直径和是否发生淋巴结转移均无相关性(P均>0.05)。WTAPP1高表达患者的5年生存率明显低于低表达者(P<0.05)。WTAPP1过表达后,SK-NEP-1细胞的增殖能力、迁移能力和侵袭能力均明显提高(P均<0.05);而沉默WTAPP1表达后,SK-NEP-1细胞的增殖能力、迁移能力和侵袭能力均明显降低(P均<0.05)。WTAPP1过表达后,SK-NEP-1细胞中Wnt3a、β-catenin、C-myc和Survivin蛋白的表达水平均明显上调(P均<0.05);沉默WTAPP1表达后,SK-NEP-1细胞中Wnt3a、β-catenin、C-myc和Survivin蛋白的表达水平均明显下调(P均<0.05)。结论:WTAPP1与肾母细胞瘤临床分期和生存预后有关,其可能通过激活Wnt/β-catenin信号通路促进瘤细胞的增殖、侵袭和迁移。

PTEN和S6K1在Ⅰ型子宫内膜癌中的表达及意义

目的探讨子宫内膜癌组织中10号染色体缺失的磷酸酶及张力蛋白同源物(PETN)和核糖体蛋白S6激酶-1(S6K1)的表达情况及其临床意义。方法选择2013年1月至2015年6月徐州医科大学附属医院及徐州市中心医院妇科行手术治疗的患者在病理科存档的组织蜡块,标本中子宫内膜癌组织65例,子宫内膜不典型增生组织30例,良性增生的子宫内膜组织30例,用免疫组化法分别检测PTEN、S6K1在不同内膜组织中的表达情况,分析两者的表达与子宫内膜癌患者各临床病理特征的相关性;对所有子宫内膜癌患者进行术后随访,采用Kaplan-Meier法进行生存分析,Cox比例风险回归模型分析子宫内膜癌患者预后的影响因素。结果(1)PTEN在子宫内膜癌组织中的阳性表达率低于正常子宫内膜组织及不典型增生子宫内膜组织(P<0.05);S6K1在子宫内膜癌组织中的阳性表达率较不典型增生子宫内膜组织、正常子宫内膜组织均增高(P<0.05)。(2)不同年龄及FIGO分期、是否淋巴结转移子宫内膜癌组织中的PTEN表达差异无统计学意义(P>0.05)。浸润深度为≥1/2肌层的PTEN阳性表达率(27.3%)显著低于无或<1/2组(59.4%),低-中分化的PTEN阳性表达率(23.3%)显著低于高分化组(60.0%)。不同年龄、不同肌层浸润深度子宫内膜癌组织S6K1阳性表达率差异无统计学意义(P>0.05)。Ⅱ~Ⅳ期、低-中分化及有淋巴结转移的S6K1阳性表达率(分别为72.4%、76.7%、81.3%)显著高于Ⅰ期、高分化及无淋巴结转移者(47.2%、42.9%、51.0%)。(3)Kaplan-Meier生存分析显示,PTEN阴性表达患者的平均生存时间短于阳性表达患者(P<0.05);S6K1阳性表达患者的平均生存时间短于阴性表达患者(P<0.05)。(4)Cox回归分析显示FIGO分期、病理分化程度、肌层浸润深度、淋巴结转移及PTEN、S6K1的表达是子宫内膜癌患者预后影响因素(P<0.05,P<0.01),其中FIGO分期、淋巴结转移及PTEN、S6K1的表达是影响子宫内膜癌预后的独立影响因素(P<0.05,P<0.01)。结论在子宫内膜癌组织中,PTEN的阴性表达和S6K1的阳性表达与子宫内膜癌的发生、发展及预后相关。