低剂量辐射对肿瘤细胞凋亡、细胞周期以及凋亡相关蛋白 Bcl-2的影响(英文)

目的探讨低剂量辐射对小鼠移植肿瘤细胞的凋亡、细胞周期以及 Bcl-2的影响。方法昆明种雄性小鼠左后肢腹股沟皮下接种 S180肉瘤细胞,接种后7天γ射线全身照射75mGy,照射后24、48小时分别处死直接测量肿瘤大小变化,取肿瘤组织分别进行流式细胞仪分析凋亡、细胞周期以及免疫组化染色半定量分析凋亡相关蛋白 Bcl-2表达的变化。结果与直接荷瘤组相比,低剂量照射组肿瘤生长缓慢(P<0.05),24小时后肿瘤细胞阻滞于 G1期,Bcl-2蛋白表达下降,48小时后肿瘤细胞凋亡增加(P<0.001)。结论低剂量辐射可使机体肿瘤细胞阻滞于 G1期并通过凋亡相关蛋白表达变化导致肿瘤细胞凋亡增加,明显提高机体抗肿瘤的作用,具有肿瘤治疗和辅助放化疗的实际临床意义。     

低剂量辐射对肿瘤细胞凋亡、细胞周期以及凋亡相关蛋白Bcl-2的影响

目的探讨低剂量辐射对小鼠移植肿瘤细胞的凋亡、细胞周期以及Bcl-2的影响。方法昆明种雄性小鼠左后肢腹股沟皮下接种S180肉瘤细胞,接种后7天γ射线全身照射75mGy,照射后24、48小时分别处死直接测量肿瘤大小变化,取肿瘤组织分别进行流式细胞仪分析凋亡、细胞周期以及免疫组化染色半定量分析凋亡相关蛋白Bcl-2表达的变化。结果与直接荷瘤组相比,低剂量照射组肿瘤生长缓慢(P<0.05),24小时后肿瘤细胞阻滞于G1期,Bcl-2蛋白表达下降,48小时后肿瘤细胞凋亡增加(P<0.001)。结论低剂量辐射可使机体肿瘤细胞阻滞于G1期并通过凋亡相关蛋白表达变化导致肿瘤细胞凋亡增加,明显提高机体抗肿瘤的作用,具有肿瘤治疗和辅助放化疗的实际临床意义。     

Notch1信号途径在食管鳞癌细胞中的激活及对细胞周期的影响

目的 观察Notch1信号途径在食管鳞癌EC9706细胞中的激活状态及 其对细胞周期的影响。方法 通过免疫细胞化学检测Notch1基因在 食管鳞癌细胞株EC9706细胞中的表达,并通过免疫荧光方法研究 Notch1基因在EC9706细胞中的激活状态。并且利用CCK-8试剂检测食管癌细胞的增殖状态。此外,采用RT-PCR和Western blot技术检 测与细胞周期相关基因的表达,最后通过流式细胞仪进一步探索激 活的Notch1信号途径对细胞周期的影响。结果 转染pcNICD后的食 管鳞癌细胞株中发现Notch1基因的表达。免疫荧光结果显示,转染 pcNICD后的食管鳞癌细胞株中Notch1信号途径处于激活状态。与未处理和转染pcDNA3.1的EC9706细胞相比,稳定表达NICD的EC9706细 胞的生长速率明显受到抑制（P<0.01）。此外,与未处理的和转染pcDNA3.1的EC9706细胞相比,稳定表达NICD的EC9706细胞的CDK2, cyclin D1和E基因的mRNA和蛋白的表达明显下调（P<0.05）。转染pcNICD的EC9706细胞在G0/G1期的比率高达74.5%,而未处理的和转染pcDNA3.1的EC9706细胞在G0/G1期的比率分别为59.1%和59.0%。细胞周期分析显示瞬时表达NICD的EC9706细胞在G0/G1期比率的增加,提示激活的Notch1信号途径能够诱导细胞静止在G₀/G₁ 期。结论 Notch1信号途径的激活引起食管鳞癌细胞的细胞周期 静止,提示Notch1基因有可能成为治疗食管鳞癌的新靶点。     

人工合成抗菌肽对口腔肿瘤细胞增殖、侵袭抑制及促凋亡作用

[目的]体外观察不同浓度的人工合成抗菌肽(artificial antimicrobial peptides,AMPs)对口腔肿瘤细胞系的增殖、侵袭抑制及促凋亡作用.[方法]合成AMPs并配置溶液,将AMPs溶液(50μg/ml～200ug/ml)加至肿瘤细胞培养体系中,收获培养后细胞以流式细胞分析法(Annexin-V/PI染色)进行细胞凋亡检测,并通过BrdU掺入实验以及MTS法进行细胞增殖检测.通过细胞迁移以及侵袭实验评价AMPs对口腔肿瘤细胞系迁移及侵袭能力的影响.[结果]AMPs可诱导口腔肿瘤细胞系SACC-83及Tca8113凋亡.在AMPs 200μmol/L浓度,中晚期凋亡比率分别为(33.89± 16.74)％、(32.47±13.53)％,高于PBS对照组的中晚期凋亡比率(4.34±1.08)％和(5.76±1.43)％(P均＜0.05),同时AMPs对肿瘤细胞促凋亡作用的效果与其浓度呈剂量依赖特点.BrdU掺人实验及MTS法发现AMPs可显著性抑制口腔肿瘤细胞系SACC-83及Tca8113的细胞增殖,当AMPs浓度为50μmol/L时对人口腔肿瘤细胞系SACC-83和Tca8113细胞增殖活性有明显的抑制作用,AMPs对人口腔肿瘤细胞系SACC-83和Tca8113细胞增殖抑制作用的IC50分别为60.38μM和55.35μM.低浓度(50μmol/L)AMPs还可显著性抑制SACC-83和Tca8113细胞的迁移以及侵袭.[结论]AMPs体外对口腔肿瘤细胞系SACC-83及Tca8113有明显的促进凋亡作用,并抑制肿瘤细胞的增殖、迁移及侵袭.     

Antitumor Efficacy of Dual Blockade of EGFR Signaling by Osimertinib in Combination With Selumetinib or Cetuximab in Activated EGFR Human NCLC Tumor Models

Introduction

Osimertinib showed great clinical efficacy for activated-EGFR NCLC patient treatment. The aim of this work was to test the efficacy of a complete EGFR-inhibition by osimertinib plus the monoclonal antibody cetuximab or the MEK1/2-inhibitor selumetinib in EGFR-mutated NCLC in vivo models.

内皮抑素对小鼠肺腺癌LA795生长和转移抑制作用的初步研究

目的：观察人内皮抑互对小鼠肺腺癌ＬＡ７９５生长和转移的抑制作用。方法：对重组人内皮抑素高效表达克隆ｐＣＸ的表达产物进行纯化,得到重组人内皮抑素（ｒｈＥＳ）。用亲和层析及胰弹性蛋白酶消化法从过期人血浆纯化得到人血管抑素（ｈＡＳ）。将接种ＬＡ７９５肺腺癌细胞的Ｔ７３９小鼠随机分成３组,分别给予ｒｈＥＳ,ｈＡＳ或等体积ＰＢＳ皮下注射,１次／日,共１４ｄ。观察３组肿瘤生长情况、肺湿重、肺表面转移结节数、动物生存期,分别进行ｑ检验。结果：ｒｈＥＳ组及ｈＡＳ组肿瘤生长缓慢,８ｄ后肿瘤逐渐回缩;肺湿重、肺表面转移结节数明显减少,动物生存期明显延长。结论：ｒｈＥＳ与ｈＡＳ均可明显抑制ＬＡ７９５所致的小鼠实验性肿瘤的生长与转移,延长动物的生存期。     

DT-5461, a New Synthetic Lipid A Analogue, Inhibits Lung and Liver Metastasis of Tumor in Mice

We have investigated the antimetastatic effect of a new synthetic lipid A analogue, of low endo-toxicity, DT-5461, against two highly metastatic tumor cell lines, L5178Y-ML25 T-lymphoma and B16-BL6 melanoma cells in mice. Four intermittent i.v. administrations of DT-5461 at intervals of 4 days resulted in a significant inhibition of liver metastasis caused by i.v. injection of L5178Y-ML25 cells and lung metastasis of B16-BL6 cells in the experimental metastasis models. Intraperitoneal and intranasal administrations as well as i.v. administration of DT-5461 were also effective in preventing lung metastasis of the melanoma cells. Multiple administrations of DT-5461 before the surgical excision of primary tumors significantly reduced the number of lung colonies of melanoma cells and primary tumor size. Similarly, this treatment modality after the surgical excision of primary tumors showed a greater reduction of lung tumor colonies as compared with lipopolysaccharide, a synthetic lipid A (No. 506) and its analogue as well as untreated control in the spontaneous lung metastasis model. Furthermore, the group that received DT-5461 after the inoculation of lymphoma or melanoma cells showed significantly enhanced survival rate compared with the untreated control. These results suggested that DT-5461 may he therapeutically useful for the inhibition of tumor metastasis.     

Sequential T Cell Response Involved in Tumor Rejection of Sarcoma, Meth A, in Syngeneic Mice

We investigated the type of T cell response involved in Meth A tumor rejection in primary immune and hyperimmune syngeneic mice. It was found that a CD4⁺ T cell-mediated delayed-type hypersensitivity (DTH) response activating non-specific killer cells such as macrophages, NK and LAK cells, without a specific CD8⁺ cytotoxic T lymphocyte (CTL) response, was the major immune response leading to Meth A tumor rejection in primary immune mice. In contrast, the specific CD8⁺ CTL response was the major response leading to the tumor rejection, in addition to CD4⁺ T cell-mediated DTH response, in hyperimmune mice. Analysis of CD4⁺ T cell clones established from primary immune and hyperimmune spleen cells indicated that a CD4⁺ T cell clone (C9) of primary immune mice (although only one clone was established) was of Th1 type, and induced cytotoxicity in accessory cells by classic DTH in vitro. Eight CD4⁺ T cell clones were established from hyperimmune spleen cells. Six out of the eight clones were of the Th2 type and two were Th0-like. However, no Th1-type CD4⁺ T cell clone was established from hyperimmune spleen cells. All of these CD4⁺ T cell clones, even the Th2-type clones, were capable of inducing cytotoxicity in vitro in T cell-depleted accessory cells, as in an in vitro DTH response. We postulate on the basis of these results that the T cell response leading to Meth A tumor rejection in vivo sequentially changed from a CD4⁺ T cell-mediated classic DTH response to a CD8⁺ CTL response, in addition to a cellular response mediated probably by Th2-type cells, during the process of repeated immunization.     

肺癌抑癌基因1对前列腺癌T_3B细胞侵袭、成瘤和转移能力的影响

[目的]研究肺癌抑癌基因1(TSLC1)对人前列腺癌T3B细胞侵袭、成瘤和转移能力的影响。[方法]将克隆有TSLC1全长cDNA的真核表达载体pCI-TSLC1稳定转染至前列腺癌T3B细胞中。实验组T3B细胞转染pCI-TSLC1质粒,以转染空质粒pCI-neo的T3B细胞为对照组,未加任何处理的T3B细胞为空白组。Transwell法检测体细胞体外侵袭能力,将三组细胞分别以4.0×106/200μl浓度注入裸鼠皮下,观察各组成瘤情况,将三组细胞分别以2.0×106/10μl进行骨原位注射建立骨转移瘤模型,观察各组骨转移率。[结果]与对照组和空白组相比,实验组细胞株细胞体外侵袭能力受到显著抑制(P<0.01);实验组皮下瘤出现明显晚于对照组和空白组,而且瘤体也明显小于对照组和空白组(P<0.01);实验组骨转移率为20%,明显低于对照组(100%)和空白组(100%)(P<0.05)。[结论]TSLC1基因明显抑制T3B细胞的侵袭、成瘤和转移能力。     

LncRNA-p21调控Notch信号通路对非小细胞肺癌A549细胞增殖、迁移及侵袭的影响

目的 探讨LncRNA-p21调控Notch信号通路对非小细胞肺癌A549细胞增殖、迁移及侵袭的影响。方法 pcDNA-lincRNA-p21、空载质粒pcDNA转染A549细胞设为过表达组和空载组;稳定转染过表达组加入Notch信号通路特异性激活剂Jagged1蛋白,设为Notch激活剂组;不作处理细胞为对照组。MTT法、划痕实验和Transwell小室实验检测各组细胞增殖、迁移和侵袭情况。RT-qPCR及Western blot法检测各组Notch1、HES-1、NICD、E-cadherin、Vimentin的mRNA和蛋白表达。结果 过表达组培养24、48和72 h MTT实验A值均低于对照组、空载组和Notch激活剂组,Notch激活剂组低于对照组和空载组（P<0.05）;过表达组48 h细胞迁移率和穿膜细胞数及Notch1、HES-1、NICD、Vimentin mRNA和蛋白相对表达量均低于对照组、空载组和Notch激活剂组,Notch激活剂组低于对照组和空载组（P<0.05）;过表达组E-cadherin mRNA和蛋白相对表达量高于对照组和Notch激活剂组,Notch激活剂组高于空载组和对照组（P<0.05）。结论 LncRNA-p21基因过表达可抑制非小细胞肺癌A549细胞增殖、迁移及侵袭,其调控机制可能与抑制Notch信号通路、阻断A549细胞上皮间质转化有关。     

Cdx2过表达对裸鼠人胃癌移植瘤生长和转移的影响

目的观察Cdx2基因过表达对裸鼠人胃癌移植瘤生长和转移的影响。方法利用脂质体将pCMV-Cdx2-HA质粒或pCMV-HA质粒分别转染人胃癌MGC-803细胞,命名为MGC-803/Cdx2细胞或MGC-803/EV细胞。将两种细胞分别注射入裸鼠近腋右背侧皮下,待皮下成瘤后将瘤组织接种于胃,建立裸鼠人胃癌移植瘤模型：接种MGC-803/Cdx2皮下瘤的裸鼠为实验组,接种MGC-803/EV皮下瘤的裸鼠为对照组;30 d后处死裸鼠,观察移植瘤生长、转移情况,并应用半定量逆转录-聚合酶链反应(RT-PCR）和Western blot技术检测移植瘤中Cdx2 mRNA和蛋白的表达。结果实验组肿瘤体积为(13.42±2.34）mm3,明显低于对照组的肿瘤体积(17.59±2.80）mm3(P<0.05）;实验组成瘤率(73.3%）低于对照组成瘤率(86.7%）;两组肿瘤转移情况比较,差异无统计学意义(P>0.05）;实验组肿瘤组织Cdx2 mRNA和蛋白表达明显增加(P<0.05）。结论 Cdx2过表达抑制裸鼠人胃癌移植瘤的生长,但对肿瘤转移无明显影响。     

Induction of Renal Cell Tumors in Rats and Mice, and Enhancement of Hepatocellular Tumor Development in Mice after Long-term Hydroquinone Treatment

Hydroquinone (HQ) was administered to F344 rats and B6C3F₁ mice of both sexes at a level of 0.8% in the diet for two years. This treatment induced renal tubular hyperplasia as well as adenomas, predominantly in males of both species, and was associated with chronic nephropathy in rats. In addition, the occurrence of epithelial hyperplasia of the renal papilla was increased in male rats. Foci of cellular alteration of the liver were significantly reduced in number by HQ in rats, but in contrast, were increased in mice, where development of hepatocellular adenoma was also enhanced in males. The incidence of squamous cell hyperplasia of the forestomach epithelium was significantly higher in mice of both sexes given HQ than in the controls, but no corresponding increase in tumor development was observed. The present study strongly indicates potential renal carcinogenicity of HQ in male rats and hepatocarcinogenicity in male mice. Thus, it is possible that HQ, which is present in the human environment, may play a role in cancer development in man.     

Association of Metastasis with Clinicopathological Data in Mexican Patients with Osteosarcoma,Giant Cell Tumor of Bone and Chondrosarcoma

Background: Bone tumors are neoplasias with a high overall mortality; one of the main factors that reduce survival is their high capacity to develop metastases. It has been reported that finding lung metastases at diagnosis of osteosarcoma (OS), chondrosarcoma (CS) and giant cell tumor of bone (GCTb) is quite common. In this study, we inquire the relationship of metastases caused by these tumors with different clinical and pathological aspects, in order to guide medical personnel in the diagnosis and opportune treatment of metastases or micro metastases. Materials and Methods: We collected data of 384 patients with clinical, radiological and histopathological diagnosis of OS, GCTb and CS that attended the National Rehabilitation Institute (INR) during 2006 to 2014. Chi-square and Fisher’s exact tests were performed for data analysis. Results: In the three tumor types, the presence of metastases at diagnosis was variable (p=0.0001). Frequency of metastases was 36.7%, 31.7% and 13.2% for OS, CS and GCTb respectively. The average age had no significant difference (p>0.05) in relation to metastases, even so, patients with OS and GCTb and metastases, were older while patients with CS and metastases were younger, in comparison to patients without metastases. Males had a higher frequency of metastases (68.2%, p = 0.09) in contrast to CS and GCTb, in which the metastases was more frequent in women with 51.9% (p = 0.44) and 57.9% (p = 0.56) respectively. Broadly, metastasis was associated with primary tumors located in the femur (44.4%), followed by the tibia (15.6%); metastases was more frequent when primary tumor of GCTb and OS were in the same bones, but were located in the hip (26.3%) for CS. Conclusions: The frequency of metastases in OS, GCTb and CS is high in our population and is determined by different clinicopathological variables related to the kind of tumor. Further studies are needed in order to evaluate metastases subsequent to diagnosis and associations with survival and clinicopathological factors , as well as to determine the sensitivity and specificity of current methods of detection.     

Hsp70-肿瘤抗原肽复合物防治小鼠黑色素瘤B16肺转移的作用

目的:探讨热休克蛋白70-肿瘤抗原肽复合物(Hsp70-antigen peptide complexes)对小鼠黑色素瘤B16转移的防治作用.方法:分别从小鼠腿部接种的B16实体瘤及小鼠肺B16转移灶提取混合抗原肽,体外与Hsp70结合制得复合物,此复合物免疫小鼠后用于预防或治疗经尾静脉接种转移至肺的B16黑色素瘤,观察其对肿瘤转移的防治作用.结果:Hsp70-肿瘤抗原肽复合物免疫后肺转移灶节结数显著减少(P＜0.01),体外脾细胞表现出对B16较高的杀伤率(P＜0.01);并对肺转移灶有显著的治疗作用(P＜0.01),而从B16实体瘤提取的混合抗原肽制得的复合物比从肺转移灶提取的混合抗原肽制得复合物有更好的治疗效果(P＜0.01),表现出体外脾细胞对B16更高的杀伤率(0.01＜P＜0.05).结论:Hsp70-肿瘤抗原肽复合物对肿瘤的转移有明显的防治作用,而从实体瘤提取的混合抗原肽比从转移灶提取的混合抗原肽更为有效.     

Inhibition of JAK1, 2/STAT3 Signaling Induces Apoptosis, Cell Cycle Arrest, and Reduces Tumor Cell Invasion in Colorectal Cancer Cells

Abnormalities in the STAT3 pathway are involved in the oncogenesis of several cancers. However, the mechanism by which dysregulated STAT3 signaling contributes to the progression of human colorectal cancer (CRC) has not been elucidated, nor has the role of JAK, the physiological activator of STAT3, been evaluated. To investigate the role of both JAK and STAT3 in CRC progression, we inhibited JAK with AG490 and depleted STAT3 with a SiRNA. Our results demonstrate that STAT3 and both JAK1 and 2 are involved in CRC cell growth, survival, invasion, and migration through regulation of gene expression, such as Bcl-2, p1^6ink4a, p21^waf1/cip1, p27^kip1, E-cadherin, VEGF, and MMPs. Importantly, the FAK is not required for STAT3-mediated regulation, but does function downstream of JAK. In addition, our data show that proteasome-mediated proteolysis promotes dephosphorylation of the JAK2, and consequently, negatively regulates STAT3 signaling in CRC. Moreover, immunohistochemical staining reveals that nuclear staining of phospho-STAT3 mostly presents in adenomas and adenocarcinomas, and a positive correlation is found between phospho-JAK2 immunoreactivity and the differentiation of colorectal adenocarcinomas. Therefore, our findings illustrate the biologic significance of JAK1, 2/STAT3 signaling in CRC progression and provide novel evidence that the JAK/STAT3 pathway may be a new potential target for therapy of CRC.     

体外内分泌治疗对乳腺癌ER细胞类型改变及其与细胞生长抑制的关系

目的：探讨乳腺癌内分泌治疗中的ER细胞类型改变的生物学意义。方法：采用体外TAM和MA单药及不同序贯或联合治疗的方法，分别作用于6组人乳腺癌Bcap-37细胞，以17荧光雌酮（17－FE）细胞化学方法观察ER在乳腺癌细胞中的分布情况。以台盼蓝拒知细胞计数法测定其细胞生长抑制率。结果：TAM单药治疗后，A型（核浆型）细胞明显增多（从3．8％升高到20．7％），并伴随C型（浆型）细胞的明显减少（从31．2％下降到4．0％），表明TAM可能使细胞内ER从细胞质向细胞核内转移；MA单药治疗时的细胞类型变化不明显，各组内分泌治疗后ER阴性细胞（即D型和E型之和）百分比相对升,设TAM→MA最高（为87．0％），MA→TAM和TAM其次（分别为79．3％和75．3％），TAM＋MA与MA相近，分别为71．3％和69．3％，而对照组为65．0％，相应的细胞生长抑制率也以TAM－→MA最高（为79．2％，P＜0．01），其次为TAM＋MA（66．5％，P＜0．01），MA最低（为40．3％），结论：TAM使细胞内ER从细胞质向细胞核内转移是本研究中1个有铁生物学现象，符合雌激素受体基因调节学说的二步工作原理，其受体细胞类型的改变与相应的抗肿瘤作用用强弱相一致，这一现象可能与ER旭性细胞被杀灭有关。     

黄芪多糖对气阴两虚Lewis肺癌荷瘤小鼠肿瘤生长、转移及细胞周期的影响

目的 观察黄芪多糖抑制气阴两虚Lewis肺癌荷瘤小鼠的生长、转移及对肺癌细胞周期的影响。方法 体外培养Lewis肺癌细胞,随机分为对照组和中药组,流式细胞术检测细胞周期;C57BL/6J小鼠90只,设空白组10只,余80只刨花烟熏并灌胃温热性的中药,移植Lewis肺癌实体肿瘤建立气阴两虚荷瘤小鼠模型并随机分为8组,比较各组小鼠抑瘤率及q值,计数外周血细胞及骨髓细胞,ELISA测定血清IL-2、IFN-γ、TNF-α、IL-10、HIF-1α、VEGF、MMP-2的含量。结果 黄芪多糖将Lewis 肺癌细胞阻滞于S期,随着药物浓度的增加,细胞凋亡逐渐增加;联合用药各组的抑瘤率高于黄芪多糖各组和顺铂组,q值均在0.85~1.15之间;与顺铂组比较,联合用药中、高剂量组小鼠外周血细胞及骨髓细胞的数量,IL-2、INF-γ、TNF-α均显著升高,IL-10、 HIF-1α、VEGF、MMP-2均显著降低（P<0.05, P<0.01）。结论 黄芪多糖在体外对Lewis肺癌细胞有抑制作用,体内与顺铂联合能抑制气阴两虚Lewis荷瘤小鼠肺癌细胞的生长、转移,提高外周血细胞和骨髓细胞的细胞数量,提高IL-2、INF-γ、TNF-α的含量,减少IL-10、HIF-1α、VEGF、MMP-2的含量。