shRNA靶向干扰COX-2促进人肝癌裸鼠皮下移植瘤肿瘤细胞凋亡

目的：探讨重组干扰质粒pshRNA-COX-2对人肝癌细胞Hep3B裸鼠皮下移植瘤肿瘤细胞凋亡的促进作用。方法：重组干扰质粒pshRNA-COX-2转染Hep3B细胞并筛选后,将被成功转染的Hep3B细胞种植于裸鼠皮下,测量肿瘤大小,成瘤4周后处死裸鼠,绘制肿瘤生长曲线,RT-PCR和Western blot检测肿瘤组...     

双氢青蒿素诱导前列腺癌PC-3细胞凋亡的形态学研究

目的：研究双氢青蒿素（DHA）诱导前列腺癌PC-3细胞凋亡作用,并观察其形态学变化。方法：以前列腺癌PC-3细胞为研究对象,在体外经不同浓度的DHA处理48小时后,扫描电镜（SEM）观察细胞凋亡;并建立PC-3细胞裸鼠种植瘤模型,20只模型鼠随机分为对照组、溶剂组、DHA大剂量组及DHA小剂量组,经13天干预后,透射电镜（TEM）观察肿瘤组织形态学变化,体视学分析肿瘤微血管密度（MVD）。结果：PC-3细胞经DHA处理后,SEM观察到典型的凋亡细胞;裸鼠种植瘤经治疗后,TEM观察肿瘤组织中散在凋亡细胞及凋亡小体,且DHA大小剂量组MVD均低于对照组（P〈0．05）。结论：DHA具有较强的抗肿瘤作用,其作用机制可能与诱导肿瘤细胞凋亡有关。     

激光光敏剂“扬州光卟啉”作用于小鼠黑色素瘤细胞疗效原理的超微结构研究

关于卟啉类物质与细胞结合的主要部位,从现有文献资料上尚无一致结论。国内实验证明国产血卟啉“扬州光卟啉”对肿瘤细胞有较强的杀伤能力。我们用透射电镜和扫描电镜研究了小鼠黑色素瘤细胞(B_(16)细胞)在“扬州光卟啉”光敏化后细胞的早期超微结构形态变化。B_(16)细胞培养在16微克/毫升和8微克/毫升“扬州光卟啉”培养液中3.5小时,再用黑光灯照射2分钟后,16微克组即可见B_(16)细胞线粒体有灶性空化。光照5分钟后,即可见B_(16)细胞肿胀,线粒体肿胀,空化及核染色质凝聚。随光照时间延长,细胞病变愈加明显,尤以线粒体病变显著。8微克组细胞也在光照5分钟后线粒体开始出现病变。这可能与“扬州光卟啉”在肿瘤细胞线粒体有很强的杀伤能力     

双氢青蒿素对前列腺癌裸鼠种植瘤CHOP表达的影响

目的：研究双氢青蒿素对前列腺癌PC-3细胞裸鼠种植瘤CHOP表达的情况,并探讨其作用途径及机制。方法：采用人前列腺癌PC-3细胞建立裸鼠种植瘤模型,20只模型鼠随机分为对照组、溶剂组、实验1组及实验2组,经13d干预后,免疫组织化学法检测CHOP的表达水平;透射电子显微镜观察DHA对PC-3细胞种植瘤超微结构的影响。结...     

C6脑胶质瘤动物模型肿瘤细胞的超微结构观察

本文对C6脑胶质瘤动物模型肿瘤标本进行了超微结构观察,以判定肿瘤生长方式及恶性程度。取成年雄性Wistar大鼠作为实验对象,采用立体定向方法将对数生长期的C6胶质瘤细胞悬液(1×106ml)注入靶点。2～3周后手术切除肿瘤并进行电镜标本制备,H600A型透射电镜观察。观察结果显示:肿瘤中心区有两种肿瘤细胞核型;不规则核型大于55%,细胞浆结构发育较差,核浆比例明显大于1。肿瘤与正常脑组织无明显边界,亦无包膜,呈浸润型生长,病理分级约有Grade～(WHO1993)。我们认为:C6细胞脑胶质瘤动物模型超微结构的观察,对进一步研究人类脑胶质瘤的恶性程度,预后判定及综合治疗具有指导意义     

胸腺上皮肿瘤的超微结构观察

目的：观察胸腺上皮肿瘤(thymic epithelial tumor,TET)WHO病理分型的超微结构特征。方法：对14例TET采用WHO病理分型,分别观察组织病理学和超微结构的形态学特征。结果：按WHO病理分型诊断胸腺瘤A型1例,AB型3例,B1型1例,B2型4例,B3型3例,C型2例。各型胸腺瘤均具有上皮性肿瘤的结构特点,如：张力原纤维、桥粒等结构;C型胸腺瘤(胸腺癌)异型性很明显,常见肿瘤性坏死,其他类型则有一定程度的异型性。A型胸腺瘤细胞核异染色质丰富,核仁不明显,无异型性;AB型和B1型异型性不明显,细胞核常染色质较均匀,可见小核仁;B2、B3型胸腺瘤异型性较明显,常染色质较丰富,核仁明显,细胞质宽大,有突起。结论：从超微结构特点来看：A型胸腺瘤生长不活跃,AB型和B1型胸腺瘤生长略活跃;B2、B3型胸腺瘤生长较活跃;C型胸腺瘤生长很活跃。     

激光诱导荧光技术在体探测裸鼠腹膜胃癌播散

以腹膜接种人胃癌细胞SGC-7901的裸鼠为胃癌腹膜播散的动物模型,模拟外科手术.在体探测不同种植期的裸鼠腹膜痛结节及止常腹膜组织的激光诱导白体荧光光谱,比对各组光谱形态,采用层次聚类分析和判别分析方法进行分类.结果表明.还原烟烟酰胺腺嘌呤二核苷酸(NADH)荧光峰强度随肿瘤细胞生长时间的增加而降低,而原卟啉IX峰逐渐升高;当类间距离的测度方法选择为欧氏距离平方,分类数取3时,可有效区分正常组、肿瘤细胞种植时间较长和较短的肿瘤组的在体荧光光谱.激光诱导自体荧光(LIAF)技术可辅助提高手术中转移癌的判断准确性.     

生长抑素对人红白血病K562细胞超微结构的影响

目的：观察生长抑素（somatostatin,SMS)对人红白血病K562细胞超微结构的影响。方法：采用体外细胞培养，加入不同浓度的8肽生长抑素（奥曲肽，octreotide)作用于K562细胞，透射电镜下观察不同浓度SMS对K562细胞超微结构影响的变化。结果：空白对照组细胞质内有各种发达的细胞器；SMS处理组随SMS浓度的增加细胞器有不同程度的变化，以10^-7mol/L-10^-6mol/L变化最明显，细胞核染色质靠核膜边集呈块状或新月形，胞浆中见核碎裂团块与空泡，线粒体基质深染、空泡样变和髓样变。结论：生长抑素可影响K562细胞的增殖，并呈现典型的细胞凋亡的超微结构变化。     

基于Ce6和MMP-2抑制剂的多级控释脂质纳米复合体用于光动力-免疫协同治疗黑色素瘤

光动力疗法（PDT）能诱导肿瘤细胞免疫原性死亡，与免疫治疗联合能够提高对各种癌症的治疗效果。基于自然杀伤（NK）细胞的免疫疗法能够增强PDT的抗癌能力。以肿瘤组织中NK细胞免疫作用的分子途径为靶点，通过药物调控增强其对肿瘤的免疫作用，并进一步联合PDT可以有效提升抗肿瘤疗效。将包封基质金属蛋白酶-2(MMP-2)抑制剂SB-3CT的β-环糊精（β-CD）与负载光敏剂Ce6的脂质体通过MMP-2响应多肽组装在一起，构建了一种肿瘤微环境/光多重响应的脂质纳米药物控制系统（MRPL-SC），在细胞和裸鼠移植瘤模型中证明了MRPLSC介导的PDT和NK免疫疗法协同抗肿瘤作用。MRPL-SC到达肿瘤部位后，连接β-CD与脂质体核心的MMP-2响应性多肽会在肿瘤组织微环境中过表达的MMP-2的作用下水解，释放包封在β-CD中的SB-3CT,SB-3CT通过直接对MMP-2进行原位抑制来有效提升肿瘤组织中NKG2DLs的表达和NK细胞免疫响应；封装了Ce6的脂质体通过细胞内吞进入肿瘤细胞后，在660 nm激光照射下通过PDT诱导肿瘤细胞凋亡，同时进一步诱导肿瘤细胞NKG2DLs表达，实现对肿瘤组织中...     

肝癌细胞黏附的压电石英谐振器实验研究

肿瘤细胞与细胞外基质(ECM)的粘附特性与肿瘤的侵袭转移密切相关,利用压电石英谐振器(QCR)实时监测HepG2肝癌细胞与ECM的黏附过程,得到频率变化(Δf)和耗散因子变化(ΔD)的响应曲线。结果表明,HepG2细胞在QCR表面经黏附、延展等行为,与ECM相互作用,此作用引起谐振晶体表面质量和粘弹性发生相应变化,Δf=(740±10)Hz,与此同时,ΔD≤5×10-5,可以认为未引起耗散因子变化,说明ECM具有刚性特征。     

RNAi沉默整合素连接激酶基因对人膀胱癌BIU-87细胞体内成瘤的影响

目的：研究RNAi沉默整合素连接激酶（integrin-linked kinase,ILK）基因对人膀胱癌BIU-87细胞裸鼠皮下成瘤的影响。方法：构建4条针对ILK基因的特异性miRNA干扰载体和1条阴性对照载体,并利用脂质体转染法将其转染BIU-87细胞,并筛选获得稳定转染细胞株。利用RT-PCR和Western blot分别从mRNA和蛋白水平检测抑制效果。将10只裸鼠随机分为2组,皮下分别接种转染组细胞和未转染组细胞,观察瘤体的生长情况,并于接种4周后处死裸鼠,测量其瘤体体积和重量,并将瘤体做HE染色,观察瘤体病理情况。结果：转染组细胞ILK的表达明显受到抑制,以miR-3组的抑制效率最高。裸鼠皮下成瘤实验检测发现：两组均有瘤体形成,未转染组较转染组瘤体生长快,未转染组体积平均为：289.56±36.49mm3,转染组为：56.67±4.32mm3。瘤体重量分别为：1.265±0.02 g和0.518±0.03g。转染组与未转染组相比差异具有统计学意义（P〈0.05）。结论：利用RNAi技术能有效抑制靶基因ILK的表达,从而降低膀胱癌细胞的体内的增殖能力。     

Enhancement of antitumor response to sarcoma 45 in rats by combination of whole-body hyperthermia and interleukin-2

AIM: To evaluate the summarized effect of hyperthermia and interleukin-2 (IL-2) administration on antitumor defense in tumor-bearing rats. METHODS: Nonbred rats after subcutaneous inoculation of sarcoma 45 cells were treated with whole-body hyperthermia (WBH, +42.5 degrees C, 60 min) and interleukin-2 (IL-2, 10,000 U/kg of body weight). Parameters of tumor growth and survival of animals were monitored till day 33 after tumor cell inoculation. RESULTS: Combined application of WBH and IL-2 at 5th day after tumor cell transplantation resulted in a delay of tumor growth and improvement of survival parameters in comparison with control group or animals that received separate treatment. Therapeutic efficacy of WBH combined with IL-2 was analogous to a single-dose chemotherapy with cyclophosphamide. CONCLUSION: Combined application of WBH and IL-2 is the useful approach for cancer immunotherapy.     

Artificial Immunogenic Cell Death Lipid Nanoparticle Functions as a Therapeutic Vaccine for Cancer

Anticancer drug-mediated induction of immunogenic cell death (ICD) blocks metastasis or recurrence in cancer cells by promoting specific immune activity against cancer antigens. However, this strategy has failed to afford adequate treatment efficiency. Overcoming the failure of ICD-mediated cancer therapy, lipid nanoparticles (LNPs) containing cancer cell surface proteins are synthesized using sonication and extrusion without microfluidics. In addition, these LNPs are decorated with high-mobility group box 1 protein and calreticulin, indicators of ICD, and named artificial ICD LNPs (AiLNPs). Administration of AiLNPs effectively targets dendritic cells (DCs) and induces DC activation in mice. Moreover, treating CT-26 tumor-bearing mice with AiLNPs inhibits tumor growth by inducing CT-26 antigen-specific T-cell immunity. Furthermore, AiLNPs containing Lewis lung carcinoma (LLC1) membrane proteins can prevent metastatic LLC1 tumor growth in the lung via LLC1 antigen-specific T-cell activation. Finally, AiLNPs synthesized with human breast cancer membrane proteins activate DC-mediated antigen-specific T-cell immunity, effectively killing tumor cells. Therefore, AiLNPs are expected to be developed as a patient-specific cancer treatment to prevent cancer recurrence and metastasis.     

近红外激光拉曼技术在体探测胃癌腹膜播散

以腹膜接种人胃癌细胞SGC-7901的裸鼠为胃癌腹膜播散的动物模型,进行模拟外科手术,在体探测不同种植期的裸鼠腹膜癌结节及正常腹膜组织的激光拉曼光谱,对比光谱差异,采用支持向量机(SVM)算法对光谱进行分类和分期判决。结果表明,癌结节和正常组织拉曼光谱差异显著,用支持向量机算法进行分类的灵敏度、特异度和诊断准确度分别为95.73%、70.73%和90.73%;不同生长期的癌结节组织拉曼光谱也存在明显差异,用支持向量机算法进行分期的结果分别为98.82%、98.73%和98.78%。从分类结果可以看出,此方法对指导外科手术中癌变组织的识别有重要的意义。     

A NIR-II Fluorescent PolyBodipy Delivering Cationic Pt-NHC with Type II Immunogenic Cell Death for Combined Chemotherapy and Photodynamic Immunotherapy

Tumor immunotherapy has emerged as one of the most promising clinical techniques to treat cancer tumors. Despite its clinical application, the cancerous immunosuppressive microenvironment limits the therapeutic efficiency of the treatment. To generate a stronger immunogenic therapeutic effect, herein, a platinum complex for chemotherapy and a BODIPY photosensitizer for photodynamic therapy are encapsulated into multimodal type II immunogenic cell death (ICD) induce nanoparticles. As the platinum complex and the photosensitizer are able to induce type II ICD, an exceptionally strong immune response is observed in triple-negative breast cancer cells. While remaining stable and therefore poorly cytotoxic in the dark, the nanomaterial is found to quickly dissociate upon exposure to near-infrared light, causing a multimodal mechanism of action in cancer cells as well as multicellular tumor spheroids through combined chemotherapy, photodynamic therapy, and immunotherapy. The nanoparticles are found to nearly fully eradicate a triple-negative breast cancer tumor and therefore to strongly enhance the survival of tumor-bearing mice models using low drug and light doses.     

光动力治疗对荷瘤小鼠肿瘤淋巴细胞表型的影响

目的:观察光动力治疗(PDT)肿瘤留置对残存肿瘤局部淋巴细胞表型的影响。方法:采用双侧小鼠荷H22移植型肝癌模型。将30只双侧荷瘤小鼠分为3组:PDT组,切除组,对照组。光动力治疗一侧肿瘤后,定量观察残存肿瘤和脾脏内CD4+和CD8+细胞的变化。使用免疫组织化学及图像分析技术。结果:在原位肿瘤内,有极少量的CD4+细胞。在残存肿瘤内,未见CD4+细胞,CD8+细胞呈明显的带状分布,在光动力治疗组,CD8+细胞的密度明显增高,但仍不足以杀死残存肿瘤,其活性也有待进一步研究。     

A Redox‐Triggered Bispecific Supramolecular Nanomedicine Based on Peptide Self‐Assembly for High‐Efficacy and Low‐Toxic Cancer Therapy

Polo‐like kinase 1 (PLK1) and polo‐like kinase 4 (PLK4) are closely associated with the progression of several cancers, and their bispecific inhibitors can kill tumor cells effectively. Herein, a redox‐responsive bispecific supramolecular nanomedicine based on the self‐assembly of a cyclic peptide, termed as C‐1, targeting both PLK1 and PLK4 as a potent anticancer agent is reported. C‐1 is a cyclic peptide in response to reducing agents such as glutathione (GSH), which is constructed by a combined approach of pharmacophore modeling, molecular docking, and reversible cyclization. After entering the cytosol of cancer cell, the disulfide linkage is reduced by intracellular GSH, with the resulting linear conformation self‐assembling into bispecific nanofibers. C‐1 can lead to apoptotic cell death by inducing caspase‐3 activation and PARP cleavage in HeLa cells. Moreover, it suppresses the growth of HeLa cells in cell assays, and inhibits the progression of HeLa cells‐induced xenografts in nude mice without inducing notable side effects. This work provides a successful example of developing the redox‐responsive bispecific nanomedicine for high‐efficacy and low‐toxic cancer therapy.