Role of autophagy in tumorigenesis,metastasis,targeted therapy and drug resistance of hepatocellular carcinoma

Autophagy is a "self-degradative" process and is involved in the maintenance of cellular homeostasis and the control of cellular components by facilitating the clearance or turnover of long-lived or misfolded proteins, protein aggregates, and damaged organelles. Autophagy plays a dual role in cancer, including in tumor progression and tumor promotion, suggesting that autophagy acts as a double-edged sword in cancer cells. Liver cancer is one of the greatest leading causes of cancer death worldwide due to its high recurrence rate and poor prognosis. Especially in China, liver cancer has become one of the most common cancers due to the high infection rate of hepatitis virus. In primary liver cancer, hepatocellular carcinoma (HCC) is the most common type. Considering the perniciousness and complexity of HCC, it is essential to elucidate the function of autophagy in HCC. In this review, we summarize the physiological function of autophagy in cancer, analyze the role of autophagy in tumorigenesis and metastasis, discuss the therapeutic strategies targeting autophagy and the mechanisms of drug-resistance in HCC, and provide potential methods to circumvent resistance and combined anticancer strategies for HCC patients.

Multifaceted p21 in carcinogenesis,stemness of tumor and tumor therapy

Cancer cells possess metabolic properties that are different from those of benign cells.p21,encoded by CDKN1A gene,also named p21Cip1/WAF1,was first identified as a cyclin-dependent kinase regulator that suppresses cell cycle G1/S phase and retinoblastoma protein phosphorylation.CDKN1A(p21)acts as the downstream target gene of TP53(p53),and its expression is induced by wild-type p53 and it is not associated with mutant p53.p21 has been characterized as a vital regulator that involves multiple cell functions,including G1/S cell cycle progression,cell growth,DNA damage,and cell stemness.In 1994,p21 was found as a tumor suppressor in brain,lung and colon cancer by targeting p53 and was associated with tumorigenesis and metastasis.Notably,p21 plays a significant role in tumor development through p53-dependent and p53-independent pathways.In addition,expression of p21 is closely related to the resting state or terminal differentiation of cells.p21 is also associated with cancer stem cells and acts as a biomarker for such cells.In cancer therapy,given the importance of p21 in regulating the G1/S and G2 check points,it is not surprising that p21 is implicated in response to many cancer treatments and p21 promotes the effect of oncolytic virotherapy.

Receptor tyrosine kinase-like orphan receptor 1:A novel antitumor target in gastrointestinal cancers

Receptor tyrosine kinase-like orphan receptor 1(ROR1)is a member of the type I receptor tyrosine kinase family.ROR1 is pivotal in embryonic development and cancer,and serves as a biomarker and therapeutic target.It has soluble and membrane-bound subtypes,with the latter highly expressed in tumors.ROR1 is conserved throughout evolution and may play a role in the development of gastrointestinal cancer through multiple signaling pathways and molecular mechanisms.Studies suggest that overexpression of ROR1 may increase tumor invasiveness and metastasis.Additionally,ROR1 may regulate the cell cycle,stem cell characteristics,and interact with other signaling pathways to affect cancer progression.This review explores the structure,expression and role of ROR1 in the development of gastrointestinal cancers.It discusses current antitumor strategies,outlining challenges and prospects for treatment.

Targeting gene-virotherapy of cancer

我们的目的到完全在动物肿瘤的异种皮移植肿瘤的消除当模特儿以便为痊愈得出 protocal 耐心。为癌症的基因治疗和病毒的治疗得到了一些治疗学的效果，但是两个都没有大突破。因此，我们得出了叫的新策略指向是基因治疗和 virotherapy 的优点的联合的癌症的 Gene-Virotherapy。这新策略独自他们比也有更强壮的反肿瘤效果。一个肿瘤特定的 replicative 侵入人体气管粘膜的病菌向量 ZD55 (E1B 55KD 删除了副词) 它类似于在指向 fuction 的 ONYX-015 但是重要在建设不同被生产，各种各样的单个治疗学的基因被插入到 ZD55。现在，象指向癌症的 Gene-Virotherapy 的如此的一个概念被提起，尽管独立 ONYX-015-tk， -cd 或 cd/-tk 等等上有一些工作，全身的联盟者以前学习了。ZD55 基因的反肿瘤效果(例如 IL-24 基因) 独自比 ZD55 (virotherapy ) 好一些并且比 Ad-IL-24 (基因治疗) 的高的百褶层独自一个。ZD55-IL-24 在在 ZD55-IL-24 治疗学习的 preclinal，完全，肿瘤质量的消除在一些老鼠然而并非在所有老鼠被发生，那意味着一基因不是足够的 effictive 在所有老鼠消除所有肿瘤团。因此，有赔偿或合作的效果的二基因独立被插入到 ZD55 并且在联合使用了。这策略被叫指向癌症的双 Gene-Virotherapy (与 % 专利) 。那么好一些的结果被获得，所有异种皮移植肿瘤群众完全在所有老鼠被消除，如果二合适的基因被选择。根据二基因结果的开始，使用二个肿瘤倡导者控制病毒向量将对指向的效果和这些药的 safty 更好，这被考虑。然后双的肿瘤控制了怀有二基因的病毒向量因为癌症治疗被得出。更好的结果被获得了，另一项专利被使用了。这反肿瘤策略能被用来与正常房间的最小的损坏在所有老鼠完全杀死所有肿瘤房间。

Therapy of cancer by cytokines mediated by gene therapy approach

基因治疗为癌症的治疗提供一条新途径。编码 immunostimulatory cytokines 的基因的转移与显著成功被使用了在动物消除癌症。然而，在有这策略的病人的临床的试用限制了功效。因此，基因转移向量系统的改进是必要的。混合病毒的向量，与鼠科的 IL-12 或记者 LacZ 基因由 SFV replicon 组成，被构造。这混合向量在 vitro 并且在 vivo 在 HCC 显示出表示的特性和高水平。在一个老鼠 orthotropic 肝肿瘤模型，没有伴随毒性，由有 mIL-12 基因的混合向量的确定的肿瘤的治疗导致了一项强壮的反肿瘤活动。随后，助手依赖者侵入人体气管粘膜的病菌包含 mifepristone (RU486 ) 的向量可诱导的系统被构造为控制并且人的 interleukin 的肝特定的表示 12 (hIL-12 )(HD-Ad/RUhIL-12 ) 并且鼠标 IL-12 (mIL-12 )(HD-Ad/RUmIL-12 ) 。数据证明 hIL-12 的高、支撑的浆液层次能被继续 RU486 的管理达到每 12 或 24 h。hIL-12 的重复正式就职能被获得在上，至少在 HD-Ad/RUhIL-12 的单个注射以后的 48 个星期的一个时期。肝转移与的处理 HD-Ad/RUmIL-12，正 RU846 在所有动物导致了完全的肿瘤回归。然后，不同 cytokine 基因被插入到有条件的 replicative 侵入人体气管粘膜的病菌向量(也叫的 oncolytic 侵入人体气管粘膜的病菌) 。在肿瘤房间的侵入人体气管粘膜的病菌的复制将杀死肿瘤房间和版本病毒，它感染包围肿瘤房间。由 oncolytic 侵入人体气管粘膜的病菌的 cytopathic 效果和 transgene 的生物效果的联合将施加强壮的反肿瘤活动。向量的这些新类型可以为癌症基因治疗提供一个有势力和安全工具。

Antitumor activity of an hTERT promoter-regulated tumor-selective oncolytic adenovirus in human hepatocellular carcinoma

AIM: To construct a tumor-selective replication-competent adenovirus (RCAd), SG300, using a modified promoter of human telomerase reverse transcriptase (hTERT). METHODS: The antitumor efficacy of SG300 in hepatocellular carcinoma was assessed in vitro and in vivo. In vitro cell viability by MTT assay was used to assess the tumor-selective oncolysis and safety features of SG300, and in vivo antitumor activity of SG300 was assessed in established hepatocellular carcinoma models in nude mice. RESULTS: SG300 could lyse hepatocellular carcinoma cells at a low multiplicity of infection (MOI), but could not affect growth of normal cells even at a high MOI. Both in Hep3B and SMMC-7721 xenograft models of hepatocellular carcinoma, SG300 had an obvious antitumor effect, resulting in a decrease in tumor volume. Its selective oncolysis to tumor cells and safety to normal cells was also superior to that of ONYX-015. Pathological examination of tumor specimens showed that SG300 replicated selectively in cancer cells and resulted in apoptosis and necrosis of cancer cells. CONCLUSION: hTERT promoter-regulated replicativeadenovirus SG300 has a better cancer-selective replication-competent ability, and can specifically kill a wide range of cancer cells with positive telomerase activity, and thus has better potential for targeting therapy of hepatocellular carcinoma.

VEGI-armed oncolytic adenovirus inhibits tumor neovascularization and directly induces mitochondria-mediated cancer cell apoptosis

脉管的 endothelial 细胞生长禁止者(VEGI ) 是肿瘤坏死因素总科的一个成员并且在脉管的动态平衡起一个重要作用。在这研究，调查 anticancer 这基因的治疗学的潜力， VEGI (VEGI-251 ) 的分泌 isoform 与 E1B 55 kDa 基因删除(ZD55 ) 被插入到有选择地复制的侵入人体气管粘膜的病菌构造 ZD55-VEGI-251。我们这里报导 potently 从 ZD55-VEGI-251-infected 癌症房间生产的分泌 VEGI-251 禁止 endothelial 房间增长，在在 vivo 的小鸡 chorioallantoic 膜的 vitro 和 angiogenesis 的试管形成。另外， ZD55-VEGI-251 感染在几根人的癌症房间线上比控制病毒导致更严重的 cytopathic 效果，包括颈的癌症房间线 HeLa， hepatoma 房间线 SMMC-7721 和 colorectal 癌症房间线 SW620。进一步的学习表明增加的 cytotoxicity 是 caspase-9 激活，提高的 caspase-3 激活和 PARP 劈开伴随的 VEGI-251 autocrine 依赖的、调停线粒体的 apoptosis 的结果。而且，显著地忍受颈的人和 colorectal 肿瘤异种皮移植的 athymic 裸体人老鼠的 ZD55-VEGI-251-treatment 压制了肿瘤生长。我们的调查结果显示 antiangiogenesis 和 apoptosis 感应的活动的联合效果为癌症使 VEGI-251-armed oncolytic 侵入人体气管粘膜的病菌成为一个有希望的治疗学的代理人。

Targeting adeno-associated virus and adenoviral genetherapy for hepatocellular carcinoma

Human hepatocellular carcinoma(HCC)heavily endangers human heath worldwide.HCC is one of most frequent cancers in China because patients with liver disease,such as chronic hepatitis,have the highest cancer susceptibility.Traditional therapeutic approaches have limited efficacy in advanced liver cancer,and novel strategies are urgently needed to improve the limited treatment options for HCC.This review summarizes the basic knowledge,current advances,and future challenges and prospects of adeno-associated virus(AAV)and adenoviruses as vectors for gene therapy of HCC.This paper also reviews the clinical trials of gene therapy using adenovirus vectors,immunotherapy,toxicity and immunological barriers for AAV and adenoviruses,and proposes several alternative strategies to overcome the therapeutic barriers to using AAV and adenoviruses as vectors.

Targeting gene-virotherapy of cancer

Due to concerns regarding the overlapping figures in this review that are identical to those contained in a reviewarticle that we have co-authored and published earlier,we retract the above paper we published in Cell Research.Weapologize for any confusion that may be caused by this matter,although we stand by the scientific contents containedin the Cell Research paper.

Local signals in stem cell-based bone marrow regeneration

骨头髓(BM ) 的细胞的基础织物开发和新生通过造血的干细胞(HSC ) 和间充质的干细胞(MSC ) 被调停。在造血的房间和 BM stromal 房间(BMSC ) 之间的本地相互影响在 myelosuppression 以后决定造血作用的体质。这里，我们在侮辱以后控制 BM 新生考察 BM 本地信号。造血的生长因素(HGF ) 和 BMSC 生产的 cytokines 是在 BM 造血作用的规定的主要因素。对在多重种类的早胚胎开发批评的 Morphogens 被加到 HSC 管理者的家庭，包括 Wnt 蛋白质，槽口 ligands， BMP，和刺猬的家庭。HSC 和 BMSC 的全球基因表示分析开始为两种房间类型揭示了基因的签名组。更重要地，生物化学、生物的结合的全球基因表示的分析在 BM 期间本地信号学习新生强烈建议了 HGF 和 cytokines 不能是为 BM 恢复的主要本地管理者， chemokines ( SDF-1 ， FGF-4 )并且 angiogenic 生长因素( VEGF --一， Ang-1 )在 myelosuppression 以后在 BM 宪法起有启发性的作用。BM 毒性的管理的一个新方向从 BM 再生管理者的鉴定正在出现。

Oncolytic viruses against cancer stem cells: A promising approach for gastrointestinal cancer

Gastrointestinal cancer has been one of the five most commonly diagnosed and leading causes of cancer mortality over the past few decades. Great progress in traditional therapies has been made, which prolonged survival in patients with early cancer, yet tumor relapse and drug resistance still occurred, which is explained by the cancer stem cell(CSC) theory. Oncolytic virotherapy has attracted increasing interest in cancer because of its ability to infect and lyse CSCs. This paper reviews the basic knowledge, CSC markers and therapeutics of gastrointestinal cancer(liver, gastric, colon and pancreatic cancer), as well as research advances and possible molecular mechanisms of various oncolytic viruses against gastrointestinal CSCs. This paper also summarizes the existing obstacles to oncolytic virotherapy and proposes several alternative suggestions to overcome the therapeutic limitations.

Effect of silk sericin on morphology and structure of calcium carbonate crystal

In this paper, silk sericin was employed to regulate the mineralization of calcium carbonate （CaC03）. CaCO3 composite particles were prepared by the precipita- tion reaction of sodium carbonate with calcium chloride solution in the presence of silk sericin. The as-prepared samples were collected at different reaction time to study the crystallization process of CaCO3 by scanning electron microscopy （SEM）, Fourier transform infrared spectroscopy （FTIR）, thermogravimetric analysis （TGA） and X-ray diffraction （XRD）. The results showed that silk sericin significantly affected the morphology and crystallographic polymorph of CaCO3. With increasing the reaction time, the crystal phase of CaCO3 transferred from calcite dominated to vaterite dominated mixtures, while the morphology of CaCO3 changed from disk-like calcite crystal to spherical vaterite crystal. These studies showed the potential of silk sericin used as a template molecule to control the growth of inorganic crystal.

Capsid modification of adeno-associated virus and tumor targeting gene therapy

Targeting is critical for successful tumor gene therapy.The adeno-associated virus(AAV) has aroused wide concern due to its excellent advantages over other viral vectors in gene therapy.AAV has a broad infection spectrum,which also results in poor specificity towards tissues or cells and low transduction efficiency.Therefore,it is imperative to improve target and transduction efficiency in AAV-mediated gene therapy.Up to now,researchers have developed many strategies to modify AAV capsids for improving targeting or retargeting only desired cells.These strategies include not only traditional chemical modification,phage display technology,modification of AAV capsid genome,chimeric vectors and so on,but also many novel strategies involved in marker rescue strategy,direct evolution of capsid proteins,direct display random peptides on AAV capsid,AAVP(AAV-Phage),and etc.This review will summarize the advances of researches on the capsid modification of AAV to target malignant cells.

Interferon-related secretome from direct interaction between immune cells and tumor cells is required for upregulation of PD-L1 in tumor cells

Dear Editor, PD-L1, also known as CD274, plays a vital role in tumor cell related immune escape. It can be expressed on the cell surface of many solid tumors （Brahmer et al., 2012） and inhibits T cell proliferation and cytokine production by bind- ing to the Tcell surface receptor programmed death 1 （PD-1） or B7-1 （McClanahan et al., 2015）. In 2013, targeting PD-1/ PD-L1 signaling for cancer immunotherapy was selected as the No.1 scientific breakthrough of the year by the editors of Science. Interferons （IFNs） are a group of pleiotropic cytokines, demonstrated anti-viral, anti-tumor, and immune regulatory functions （York et al., 2015）. Type I interferon binds a heterodimeric receptor composed of IFNAR1 and IFNAR2. This activates a canonical JAK/STAT signaling pathway that ultimately induces a set of interferon-stimulated genes to exert its biological activity （Ejlerskov et al., 2015）. Recently, PD-L1 was reported to be downstream of IFN signaling in human oral squamous carcinoma, melanoma, and human acute myeloid leukemia blast cells （Chen et al., 2012; Furuta et al., 2014; Kronig et al., 2014）.

SNORD126 promotes HCC and CRC cell growth by activating the PI3K-AKT pathway through FGFR2

小核仁的 RNA (snoRNA ) 机能障碍与癌症开发被联系了。SNORD126 是在它的主机基因的 introns 5-6 以内被编码的孤儿 C/D 框 snoRNA， cyclin 交往 B1 蛋白质 1 (CCNB1IP1 ) 。SNORD126 触发的联系癌症的分子的机制充分没被理解。这里，我们证明 SNORD126 高度在 hepatocellular 癌(HCC ) 被表示， colorectal 癌症(CRC ) 病人取样。SNORD126 在裸体老鼠增加了 Huh-7 和 SW480 细胞生长和 tumorigenicity。SNORD126 击倒禁止的 HepG2 和 LS174T 房间生长。我们证实 SNORD126 没与 miRNA 活动被处理成小 RNA。而且， SNORD126 没在 HCC 样品与 CCNB1IP1 显示出重要表示关联或调整 CCNB1IP1 表示。我们的基因表示侧面分析显示了基因经常印射到 PI3K-AKT 小径的那 SNORD126-upregulated。SNORD126 overexpression 增加了 phosphorylated AKT， GSK-3，和 p70S6K 的层次并且提高了成纤维细胞生长因素受体 2 (FGFR2 ) 表示。在 SNORD126-overexpressing Huh-7 细胞的 FGFR2 的调停 siRNA 的击倒或调停 AZD4547 的 inactivation 禁止了 AKT phosphorylation 并且压制了细胞生长。这些调查结果在癌症为 SNORD126 显示一个 oncogenic 角色并且作为一个治疗学的目标建议它的潜力。

Preparation of 3-D porous fibroin scaffolds by freeze drying with treatment of methanol solutions

In this study, silk scaffolds with appropriate porous structures were prepared by adjusting solution concentrations and providing treatment with methanol solutions in the way of freeze drying. The effects of the preparation conditions on the microstructures and properties of the scaffolds were discussed. Fibroin solutions with different concentrations of 4, 6, 8, 10 wt% were used respectively to prepare the scaffolds. The effects of the addition of 20 vol% methanol before or after freeze drying to the 4 wt% fibroin solution were investigated. As demonstrated by Scanning Electron Microscope (SEM), the fib-roin scaffolds prepared without methanol had porous microstructures composed of thin sheets, and the sizes of the pores decreased with the increase of the fibroin solution concentrations, while the scaffolds prepared in the presence of methanol showed porous microstructures formed by fine-particle aggregates. The porosities and mechanical properties of the prepared fibroin scaffolds under different conditions were tested. The crystalline structures and conformations of the fibroin scaffolds were detected by Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD).

Targeting strategies for adeno-associated viral vector

Adeno-associated virus (AAV) has rapidly become a promising gene delivery vector for its excellent advantages of low pathogenicity and long-term gene expression over other viral vectors for human gene therapy. However, the disadvantage of AAV for clinical gene therapy is the broad host range and the lack of tissue or cell specificity, which makes AAVs relatively inefficient gene delivery to target cells and short of safety. Therefore, it is required to improve the targeting capability of recombinant adeno-associated virus (rAAV) for successful gene therapy. At present, the successful identification of vari- ous-serotype AAVs and the improvement of rAAV production technology make it feasible to explore new applications of rAAV vector in clinical gene therapy trials. Moreover, the current development in targeting strategy has brought a bright prospect to rAAV vectors. Many studies have demonstrated that targeting gene therapy based on rAAV vectors could be achieved through transcription targeted, con-jugate targeted, receptor targeted, gene recombinant targeted strategies, etc. This report gives a com- prehensive review of the recent developments of targeting strategies for rAAV vectors.

Scalable manufacturing methodologies for improving adeno-associated virus-based pharmaprojects

Adeno-associated virus(AAV)is a promising viral vector and meets most requirements of being a safe biological agent.However,the commercialization of AAV has been hampered due to the limitation of large-scale production,and only a small number of clinical trials have been launched.In recent years,progresses in scalable manufacturing of AAV have dramatically improved AAVbased clinical researches,and have assisted the development of investigational drug products.An AAV1-based investigational product,Glybera,has been formally approved by European Commission for the treatment of lipoprotein lipase deficiency(LPLD).Glybera was the first gene therapy product in the western world,and the production process involves a scalable baculovirus-insect cell system.However,many problems still need to be solved to improve the productivity and quality of AAV.The present review gives critical insights into current state-of-the-art scalable producing methodologies of AAV,such as baculovirus-insect cell system,HSV complementation system,and Ad complementation system,along with a discussion on the problems,solutions,and developmental trends.Novel AAV-producing platforms in Saccharomyces cerevisiae and vaccinia virus complementation system will also be discussed.