125I标记的Flt4多抗检测荷瘤小鼠前哨淋巴结的探讨

目的 探讨125I标记的Flt4多抗(125I-Flt4PcAb)对荷瘤小鼠前哨淋巴结(sentinel lymph node,SLN)的检测,为应用Flt4PcAb进行SLN特异性定位提供实验依据.方法 建立BALB/c裸小鼠后肢荷瘤模型,7周后应用125IFh4PcAb检测肿瘤的SLN,健侧作为对照,切取(月园)窝淋巴结探测γ射线的每分钟计数率(count per minute,Cpm),并进行HE及角蛋白免疫组化染色;分析不同状态淋巴结摄取125I-Flt4PcAb的特点.结果 BALB/c裸小鼠后肢皮下注射Tca-8113细胞悬液,移植瘤成功率达100%;患侧70枚(月园)窝淋巴结中,HE染色3枚(4.3%)出现肿瘤转移;角蛋白免疫组化染色为5枚(7.1%);125I-Flt4PcAb摄取方面,肿瘤转移淋巴结、反应性增生淋巴结和正常淋巴结之间均有显著差异(P＜0.05).结论 BAIB/c裸小鼠后肢皮下注射Tca-8113细胞悬液可成功致瘤,但淋巴结转移率低;免疫组化染色较HE染色检测肿瘤转移更敏感;125I-Flt4PcAb能够检测到荷瘤鼠的SLN,肿瘤转移淋巴结对125I-Flt4PcAb的摄取下降.     

~(125)Ⅰ标记的Flt4多抗检测荷瘤小鼠前哨淋巴结的探讨

目的探讨125I标记的Flt4多抗(125I-Flt4PcAb)对荷瘤小鼠前哨淋巴结(sentinel lymph node,SLN)的检测,为应用Flt4PcAb进行SLN特异性定位提供实验依据。方法建立BALB/c裸小鼠后肢荷瘤模型,7周后应用125I-Flt4PcAb检测肿瘤的SLN,健侧作为对照,切取窝淋巴结探测γ射线的每分钟计数率(count per minute,Cpm),并进行HE及角蛋白免疫组化染色;分析不同状态淋巴结摄取125I-Flt4PcAb的特点。结果BALB/c裸小鼠后肢皮下注射Tca-8113细胞悬液,移植瘤成功率达100%;患侧70枚窝淋巴结中,HE染色3枚(4.3%)出现肿瘤转移;角蛋白免疫组化染色为5枚(7.1%);125I-Flt4PcAb摄取方面,肿瘤转移淋巴结、反应性增生淋巴结和正常淋巴结之间均有显著差异(P<0.05)。结论BALB/c裸小鼠后肢皮下注射Tca-8113细胞悬液可成功致瘤,但淋巴结转移率低;免疫组化染色较HE染色检测肿瘤转移更敏感;125I-Flt4PcAb能够检测到荷瘤鼠的SLN,肿瘤转移淋巴结对125I-Flt4PcAb的摄取下降。     

两种不同淋巴转移能力小鼠肝癌细胞亚系的建立和生物学特性

通过单细胞分离(克隆)培养和局部淋巴结转移灶筛选的方法,建立两种不同淋巴转移能力HepA肝癌细胞亚系,分别命名为HepA—H和HepA—L,并比较其生长、游走和贴壁能力等生物学特性。瘤细胞接种于小鼠足垫后,同侧膕窝淋巴结转移率,HepA—H为83．3％,HepA—L为16．7％,两者具有显著差异(p＜0．01)。两种细胞亚系均未见肺、肝、脾、肾等脏器转移灶。体外实验显示,HepA—H的生长、游走和贴壁能力均强于HepA-L。HepA肝癌不同淋巴转移能力亚系的建立,为研究肿瘤经淋巴管转移机理提供了良好的肿瘤淋巴转移动物模型。     

两种不同淋巴转移能力小鼠肝癌细胞亚系的建立和生物学特性

通过单细胞分离(克隆)培养和局部淋巴结转移灶筛选的方法,建立两种不同淋巴转移能力HepA肝癌细胞亚系,分别命名为HepA-H和HepA-L,并比较其生长、游走和贴壁能力等生物学特性。瘤细胞接种于小鼠足垫后,同侧腘窝淋巴结转移率,HepA-H为83.3%,HepA-L为16.7%,两者具有显著差异(p<0.01)。两种细胞亚系均未见肺、肝、脾、肾等脏器转移灶。体外实验显示,HepA-H的生长、游走和贴壁能力均强于HepA-L。HepA肝癌不同淋巴转移能力亚系的建立,为研究肿瘤经淋巴管转移机理提供了良好的肿瘤淋巴转移动物模型。     

裸鼠体内高转移人肺癌模型的筛选

将人肺巨细胞癌ＰＬＡ－８０１Ｄ裸鼠皮下移植瘤作为瘤源,建立了ＰＬＡ－８０１Ｄ－ＡＳ裸鼠腹水鼠模型,其转移率及转移程度明显增高;以ＰＬＡ－８０１Ｄ－ＡＳ腹水瘤模型的肺转移灶细胞进行裸鼠皮下连续传代,建立了肺转移率及程度高于ＰＬＡ－８０１Ｄ的ＰＬＡ－８０１ＤＬ皮下移植瘤株。本文从肿瘤的异质性以及肿瘤与宿主相互作用的角度讨论了其肺转移提高的原因。     

小鼠Lewis肺癌不同部位皮下移植瘤模型的比较

目的探究不同部位肺癌皮下移植瘤存在的差异,为肺癌研究者提供提供基础依据。方法将稳定表达虫萤光素酶的小鼠Lewis肺腺癌细胞(ll2-luc-m38)分别注射于C57 BL/6小鼠的右腋皮下、右腹股沟皮下、脚垫皮下,定期利用小动物活体成像系统观察小鼠皮下移植瘤成瘤情况及转移情况,观察不同部位荷瘤小鼠的生存时间和死亡率,肿瘤组织取材,采用石蜡包埋、切片、HE染色做出病理学诊断。并将小鼠处死后立即进行肺部组织固定、观察转移灶。对皮下移植瘤行切除术观察小鼠术后生存情况和转移情况。结果腋下组和腹股沟组成瘤时间较早,成瘤率为100%,脚垫组成瘤时间较晚,成瘤率为33%;腹股沟组和腋下组瘤体积增长迅速,其中腹股沟组瘤体积增长最快;接种后第21天腋下组70%的小鼠出现了肺转移,转移灶数目较多;腹股沟组50%小鼠出现肺转移,且转移灶数目较少;脚垫组小鼠未观察到肺转移灶。脚垫组小鼠死亡率最高。腹股沟组和腋下组小鼠可行皮下移植瘤切除术,术后存活率为100%。结论小鼠lewis肺癌皮下移植瘤模型中,腹股沟组和腋下组成瘤率高,可耐受移植瘤切除术,手术死亡风险低,便于监测,操作简单且可重复性高,其中腋下组转移性更好。     

人乳腺癌MCF-7细胞在放射线处理的SCID小鼠中转移的实验研究

目的建立人乳腺癌MCF-7细胞SCID(Severe combined immunodeficiency,SCID)小鼠转移动物模型。方法采用人乳腺癌细胞株MCF-7细胞悬液,分别接种于5只经放射线处理的SCID小鼠腋背部皮下。记录肿瘤生长情况,处死荷瘤鼠并做病理切片,观察各脏器转移情况。结果接种SCID小鼠后6～10d成瘤,成瘤率为5/5只,潜伏期平均(7.4±1.3)d。接种后5只鼠分别于第60～68天拉颈处死,检测荷瘤,平均直径为(26.6±2.2)mm,平均重量为5.28g。病理学检查,转移脏器有3个部位,出现肺转移的为4/5只、骨转移的为3/5只和淋巴结转移的为1/5只。结论建立了人乳腺癌SCID小鼠转移动物模型,该模型可为肿瘤转移研究提供重要的实验工具。     

A549人肺癌细胞系/615-SCID小鼠转移瘤的生物学特征

目的通过建立A549人肺腺癌细胞/615-SCID小鼠模型,评价重度联合免疫缺陷615-SCID小鼠在建立人类肺癌转移模型方面的应用价值.方法将1×107A549细胞接种到615-SCID及SCID小鼠右上肢背部皮下,观察成瘤时间、成瘤率、肿瘤生长速度及转移发生.结果两品系小鼠接种后的成瘤率均为100%,615-SCID小鼠移植瘤潜伏期较长、生长较缓慢,更容易发生转移.结论 615-SCID小鼠比SCID小鼠更易于构建人类肺腺癌转移模型,对于肺癌转移特性研究具有较大的意义.     

GFP标记的肺癌细胞生长和转移的在体荧光成像

目的:利用稳定表达GFP的人高转移肺巨细胞癌细胞株95D,在体观察肿瘤的生长和淋巴结转移情况.方法:利用逆转录病毒感染人高转移肺巨细胞癌细胞株95D,经过G418筛选获得稳定表达GFP的细胞株.接种裸鼠皮下,成瘤后使用Nikon公司SMZ1000 P-FLA型荧光体视显微镜观察皮下原发肿瘤病灶及转移淋巴结.结果:使用逆转录病毒感染,可以获得稳定表达高强度GFP的肺癌细胞株,且生物学行为无明显改变.接种至裸鼠皮下后,2周左右成瘤,皮下肿瘤原发病灶在荧光体视镜下可清晰观察到肿瘤侵犯范围及周边血管生长情况.切开皮肤,可以观察到表达的GFP的转移淋巴结.结论:GFP标记人高转移肺巨细胞癌细胞株95D,建立裸鼠移植瘤模型,能更方便的观察肿瘤的生长、侵润和转移,为研究肿瘤的生长转移机制打下基础.     

骨髓间充质干细胞全细胞抗原干预移植瘤生长的实验研究

目的:探讨骨髓间充质干细胞(Mesenchymal Stem cells,MSCs)的全细胞抗原(WCAs)对于人肺腺癌细胞株A549移植瘤的预防接种作用,为发掘肿瘤免疫治疗提供新策略。方法:采用全骨髓贴壁法原代培养小鼠MSCs并以流式细胞仪鉴定,取3~5代MSCs细胞以15Gy X线灭活获取(Whole cell antigens,WCAs),以该抗原皮下接种于BALB/c小鼠(1次/3 d,共2周),获得免疫接种小鼠模型,对照组皮下注射同体积的PBS;为了方便示踪,以Lipofectamine TM 2000细胞脂质体转染绿色荧光蛋白(GFP),获得标记GFP-A549细胞株,皮下移植瘤株进行荷瘤,采用小动物荧光成像仪观察肿瘤生长;同时评估肿瘤直径并计算肿瘤体积,行Ki-67免疫组化染色初步分析肿瘤增殖能力。结果:肺腺癌GFP-A549细胞株皮下移植成功使小鼠荷瘤,实验组小鼠的肿瘤体积显著小于对照组(P0.05,P0.01),小动物荧光成像仪结果与解剖结果一致;实验组Ki-67的表达低于对照组。结论:本研究采用MSCs获得WCAs在荷瘤前对小鼠进行免疫刺激,发现其确实有抑制肿瘤生长的作用。     

胃癌的肿瘤微环境研究进展

肿瘤微环境是决定肿瘤细胞行为的主要影响因素,有别于正常细胞与其周围组织所形成的微环境,组织缺氧和酸中毒、间质高压形成、大量生长因子和蛋白水解酶的产生及免疫炎性反应等构成了肿瘤组织代谢环境的生物学特征,这种特性在肿瘤的发生、进展、转移中扮演重要的角色。胃癌早期症状不典型、转移迅速、死亡率高,是消化系统最常见的恶性肿瘤,目前,关于肿瘤微环境的研究尚处于起步阶段,对胃癌肿瘤微环境的研究有助于我们进一步认识胃癌发生发展的机制,并为临床诊断、治疗胃癌提供依据。因此,本文就近年来在胃癌肿瘤微环境方面的研究进展作一综述。     

Gamma- 突触核蛋白与肿瘤关系的研究进展

Synucleins家族是一种主要在神经元内表达的高度可溶性的蛋白家族。已知synucleins家族有3个成员α-synuclein(SNCA),β-synuclein(SNCB)和γ-synuclein(SNCG)。其中SNCA与SNCB参与神经递质释放过程,被认为与神经退行性变疾病密切相关,特别是在阿尔茨海默病与帕金森病中常有异常表达。近年来研究表明,SNCG在人类许多肿瘤中过表达,如乳腺癌、结直肠癌、女性生殖系统肿瘤、前列腺癌、膀胱癌等。通过研究表明γ-Synucleins异常表达机制目前包括DNA甲基化、激动蛋白-1激活、对转录因子SP1结合位点、有丝分裂检验点基因、雌激素受体影响。通过以上机制,SNCG促进上述肿瘤细胞的增殖、抑制肿瘤细胞的凋亡以及促进肿瘤细胞的转移。根据文献分析,我们提出SNCG可作为多种肿瘤预后的潜在指标的可能,同时以SNCG为切入点,探讨神经系统是否可以对肿瘤发生、发展的影响作用。     

Effect of cisplatin upon expression of in vivo immune tumor resistance

The major intent of cancer treatment with cytotoxic drugs is direct tumor cell damage, but some of these drugs have been shown to be immunomodulatory. Cisplatin is a widely used cytotoxic drug that has been combined with biological response modifiers in recent clinical trials. To evaluate further whether cisplatin may independently alter the level of host resistance against tumor growth, the drug was tested in the Mc7 sarcoma rat tumor model. The expression of in vivo tumor resistance against Mc7 sarcoma in syngeneic Wistar rats is mediated by circulating non-cytotoxic T lymphocytes. These cells interact specifically with tumor cells to generate cytotoxic effectors locally at the site of a tumor challenge. Activities of these components of expression of tumor resistance were measured in vivo after administration of cisplatin and dose-dependent effects were found. Low-dose cisplatin (0.3 mg/kg) increased the activity of the circulating lymphocytes that mediate tumor resistance, and high-dose cisplatin (9 mg/kg) suppressed both mediator lymphocyte activity and the generation of antitumor effector mechanisms. These studies suggest that low-dose cisplatin may be immunomodulatory and combining it with biological response modifiers might be a useful strategy. However, high-dose cisplatin given with biological response modifiers may negate potential immunomodulatory activities of such agents.     

Biochemical Analysis of Metastasis-Related Ax Actin in B16 Mouse Melanoma Cells After Chemical Reversional Modulation and of Tumor Progression-Related A′ Actin in the Ontogeny of Human Malignant Melanoma

To examine the correlation between tumor metastasis and A^x actin in mouse melanoma and between tumor progression and A′, actin in human melanoma and further to investigate whether or not it is a generally existing principle, we studied the effects of reversion agents, which distinctly decrease metastatic ability of melanoma cells, on the appearance of A^x actin. Will an induced decrease in metasasis of established highly metastatic B16-F10 mouse melanoma cells cause the appearance of A^x actin? We also examined the appearance of A′ actin in eight human benign pigment cell tumors and nine human malignant melanoma tissues or cells in relation to tumor progression. In vitro treatment of B16-F10 cells with each of these agents suppressed metastatic ability of the cells injected intravenously into syngenic mice; however, none of the treated cells represented A^x actin in vitro. These results suggest that the appearance of A^x actin may be a result of long-term tumor cell progression leading to changes in gene level, but because the treatments with these agents were only carried out over a short period, they could not effect changes in gene level; thus, A^x actin appearance remained unchanged. Appearance of A′ actin was detected only in human benign pigment cell tumors such as nevus cell nevi, but not in malignant melanomas, which were also formed in a long period of tumor progression in vivo. These results suggest that A′ actin is a clinically useful marker to determine the prognosis and level of tumor progression of human pigment cell tumors.     

TNF-a在肿瘤中的作用

肿瘤坏死因子是一个特别的,并具有多重功能的细胞因子,它在免疫调节,炎症反应,机体防御当中起着关键的作用。根据不同的细胞微环境,肿瘤坏死因子可以诱导多种反应,例如凋亡,坏死,血管生成,免疫细胞激活,细胞分化,细胞迁移。TNF在肿瘤当中是一把双刃剑。一方面,TNF是一个内源性的肿瘤促进因素,因为TNF可以刺激肿瘤细胞生长,增殖,侵袭,转移,血管生成。另一方面,TNF具有杀肿瘤细胞的作用。因此,如果可以调控肿瘤坏死因子的功能,将为癌症的治疗提供可能。     

Tumor-suppressive Function of Protein-tyrosine Phosphatase Non-receptor Type 23 in Testicular Germ Cell Tumors Is Lost upon Overexpression of miR142–3p microRNA

Protein-tyrosine phosphatase non-receptor type 23 (PTPN23) is a candidate tumor suppressor involved in the tumorigenesis of various organs. However, its physiological role(s) and detailed expression profile(s) have not yet been elucidated. We investigated the function and regulation of PTPN23 in the formation of testicular germ cell tumors (TGCTs). Expression of PTPN23 in human TGCT cell lines was significantly lower than that in spermatogonial stem cells in mice. Overexpression of PTPN23 in NEC8, a human TGCT cell line, suppressed soft agar colony formation in vitro and tumor formation in nude mice in vivo. These data indicate that PTPN23 functions as a tumor suppressor in TGCTs. Multiple computational algorithms predicted that the 3′ UTR of human PTPN23 is a target for miR-142-3p. A luciferase reporter assay confirmed that miR-142-3p bound directly to the 3′ UTR of PTPN23. Introduction of pre-miR-142 in the PTPN23 transfectant of NEC8 led to suppressed expression of PTPN23 and increased soft agar colony formation. Quantitative RT-PCR data revealed a significantly higher expression of miR-142-3p in human seminomas compared with normal testes. No difference in mRNA expression between seminoma and non-seminoma samples was detected by in situ hybridization. Both quantitative RT-PCR and immunohistochemical analyses revealed that PTPN23 expression was significantly lower in TGCTs than in normal testicular tissues. Finally, a lack of PTPN23 protein expression in human TGCTs correlated with a relatively higher miR-142-3p expression. These data suggest that PTPN23 is a tumor suppressor and that repression of PTPN23 expression by miR-142-3p plays an important role in the pathogenesis of TGCTs.     

Aberrant leukocyte infiltration: a direct trigger for breast tumor invasion and metastasis

Our previous studies revealed that leukocyte infiltration could trigger breast and prostate tumor invasion through physical disruption of tumor capsules. Our current study, involving multiple types of human tumors, further suggests that leukocyte infiltration also triggers metastasis through the following pathways : 1) the physical movement into the epithelium disrupts inter-cellular junctions and surface adhesion molecules, which cause the disassociation of tumor cells from tumor cores, 2) some of these tumor cells subsequently form tight junctions with the plasma membranes of leukocytes creating tumor cell-leukocyte chimeras (TLCs), and 3) the leukocytes of TLCs impart migratory capacity to associated tumor cell partners. Our findings suggest a novel pathway for tumor cell dissemination from primary sites and journey to new sites.     

Functional mechanism of Ginsenosides on tumor growth and metastasis

Ginsengs, has long been used as one medicinal herb in China for more than two thousand years. Many studies have shown that ginsengs have preventive and therapeutic roles for cancer, and play a good complementary role in cancer treatment. Ginsenosides, as most important constituents of ginseng, have been extensively investigated and emphasized in cancer chemoprevention and therapeutics. However, the functional mechanism of Ginsenosides on cancer is not well known. This review will focus on introducing the functional mechanisms of ginsenosides and their metabolites, which regulate signaling pathways related with tumor growth and metastasis. Ginsenosides inhibit tumor growth via upregulating tumor apoptosis, inducing tumor cell differentiation and targeting cancer stem cells. In addition, Ginsenosides regulate tumor microenvironment via suppressing tumor angiogenesis-related proteins and pathways. Structural modification of ginsenosides and their administration alone or combinations with other Chinese medicines or chemical medicines have recently been developed to be a new therapeutic strategy for cancer.     

A novel immunoregulatory axis of NKT cell subsets regulating tumor immunity

There are many mechanisms that regulate and dampen the immune response to cancers, including several types of regulatory T cells. Besides the T reg cell, we have identified another immunoregulatory circuit initiated by NKT cells that produce IL-13 in response to tumor growth and this IL-13 then induces myeloid cells to make TGF-beta that inhibits cytotoxic T cell-mediated tumor immunosurveillance in several mouse tumor models. This finding created a paradox in the role of NKT cells in tumor immunity, in that they can also contribute to protection. We resolve this paradox by the finding that the suppressive NKT cell is a type II NKT cell that lacks the canonical invariant T cell receptor, whereas the protective cell is a type I NKT cell that expresses the invariant receptor. Further, we see that these two subsets of NKT cells counter-regulate each other, defining a new immunoregulatory axis. The balance along this axis may determine the outcome of tumor immunosurveillance as well as influence the efficacy of anti-cancer vaccines and immunotherapy.     

腹腔注射精氨酸对小鼠S_（180）瘤生长的促进作用

尽管大多数动物实验的结果表明通过饮水或饲料补充精氨酸对一些肿瘤的形成、生长及转移均有显著的抑制作用,但也有少数几篇相反结果的报道。我们采用小鼠腹腔注射的方法,观察了不同剂量（０．１、０．０５、０．０２５克／每天）精氨酸对Ｓ１８０瘤生长的影响,结果发现精氨酸对Ｓ１８０瘤的生长具有显著的促进作用,尤其是０．０５克／每天剂量组,文中就其可能机制进行了讨论。