髓源性抑制细胞和调节性T细胞在感染和肿瘤微环境中的免疫抑制机制

髓源性抑制细胞(MDSC)和调节性T细胞(Treg)是重要的抑制性免疫细胞,在炎症、感染和肿瘤中大量扩增,可通过多种机制抑制机体抗肿瘤免疫,促进肿瘤生长和转移.寻找肿瘤微环境中MDSC、Treg细胞升高的原因及清除方法,已成为肿瘤免疫治疗的研究热点.     

Photodynamic therapy for breast cancer in a BALB/c mouse model

Objective

Photodynamic therapy (PDT) has been used for superficial neoplasms and its usage has been recently extended to deeper lesions. The purpose of this study was to observe whether or not PDT can cure breast cancer in the solid tumor model, and to define the critical point of laser amount for killing the cancer cells.

Methods

Twenty four BALB/c mouse models with subcutaneous EMT6 mammary carcinomas were prepared. Mice were divided into eight groups depending on the amount of illumination, and the tumor size was between 8 mm and 10 mm. We began by peritoneal infiltration with a photosensitizer 48 hours prior to applying the laser light, and then we applied a non-thermal laser light. The energy was from 350 J/cm² to 30 J/cm² to the cancer.

Results

Regardless of the tumor size from 8 mm to 10 mm, all mice apparently showed positive results via PDT. We also did not find any recurrence over 90 J/cm². In all models, the color of the breast cancer lesions began to vary to dark on 2 days post PDT and the tumor regression began simultaneously. Also, we confirmed the complete regression of the breast cancer 21 days after PDT.

Conclusion

We confirmed that PDT may treat breast cancers that are sized less 10 mm in mouse models. The moderate energy to destruct the breast cancer cells may be 90 J/cm². Therefore, we can expcect that PDT may be utilized to treat breast cancer, but we need more experience, skills and processing for clinical trials.     

BALB／C小鼠黑色素瘤肿瘤模型的建立及对荷瘤鼠的影响

目的建立小鼠不同部位黑色素瘤模型，为不同研究目的提供合适的动物肿瘤模型。方法接种B16小鼠黑色素瘤细胞于BALB／C小鼠足部、腹腔和尾静脉，建立移植瘤模型和肺部转移瘤模型。观察B16实体瘤的形态学特点，接种细胞数与肿瘤生成的关系，肿瘤生长对小鼠生存时间、血清肿瘤坏死因子（sTNF）和唾液酸（SA）的影响。结果足部接种5X10瘤细胞后10天，肿瘤进入对数生长，成瘤率近100％；腹腔接种瘤细胞数大于5000个／只，成瘤率与接种细胞数成正比；接种3X10细胞／只，10天后在小鼠腹壁和肠系膜可见散在黑色素瘤结节，荷瘤鼠sTNF和SA水平明显高于正常小鼠，且与肿瘤的大小和数量成正比，小鼠平均存活59天；尾静脉接种后3周，肺部可见大小不等转移瘤灶，两肺平均瘤灶数可达192.11士92。结论B16黑色素瘤细胞株在BALB／C小白鼠具有很高的成瘤率和转移特性，且较C57／B16小黑鼠模型具有方便、稳定和观察容易等优点，是一较理想的模型。     

蒿甲醚对BALB／c小鼠CT-26结直肠癌抑瘤作用

[目的]探讨蒿甲醚(Artemether)对BALB/c小鼠CT-26结直肠癌的抑瘤作用。[方法]BALB/c小鼠皮下接种CT-26结直肠癌细胞(2×106)48只,雌雄各半;随机分为6组,每组8只,分别为低剂量(33.3mg/kg)、中剂量(50mg/kg)、高剂量(66.6mg/kg)和中剂量(50mg/kg)+铁剂(1.5mg/kg)组,阳性对照组为顺铂(5mg/kg),空白对照组为等体积生理盐水。除顺铂为腹腔注射外,其余组均为灌胃给药法。[结果]口服蒿甲醚低、中、高剂量和中剂量+铁剂对BALB/c小鼠CT-26结直肠癌的抑瘤率分别为:42.3%、51.4%、52.0%、53.5%。[结论]在一定剂量范围内,口服蒿甲醚对小鼠CT-26结直肠癌有明显的抑制作用;蒿甲醚与铁剂合用具有一定的协同作用。     

Myeloid-Derived Suppressor Cells in Nonmetastatic Urothelial Carcinoma of Bladder Is Associated With Pathologic Complete Response and Overall Survival

BackgroundMyeloid-derived suppressor cells (MDSC) have immunosuppressive activity and enhance tumor progression. We hypothesized that lower blood MDSC would correlate with pathologic complete response and better outcomes in nonmetastatic urothelial carcinoma (UC).Patients and MethodsBefore cystectomy, blood MDSC were measured in whole blood (WB) and peripheral blood mononuclear cells using flow cytometry. MDSC were defined as CD33⁺/HLA-DR⁻. MDSC subtypes were polymorphonuclear MDSC (CD15⁺/CD14⁻), monocytic (M)-MDSC (CD15⁻/CD14⁺), and uncommitted (UnC) MDSC (CD15⁻/CD14⁻). The Wilcoxon rank sum test was used to compare MDSC between pathologic complete response groups. The optimal cutoff points for MDSC were identified using recursive partitioning analysis with cross-validation. The Cox proportional hazard model was used to associate MDSC and other clinical factors with recurrence-free survival and overall survival (OS).ResultsOverall, 109 patients were included: 86% men with median (range) age of 67 (30-88) years, 76% with pure UC, 29% intravesical therapy, and 41% neoadjuvant chemotherapy. Twenty-one patients (19%) had pT0N0 and 23 (24%) < pT2N0. Median (range) follow-up time was 17.4 (0.4-42.4) months. Total MDSC and polymorphonuclear MDSC percentage in peripheral blood mononuclear cells was significantly lower in patients with pT0N0 disease (P = .03). One- and 2-year OS rates were 94% (95% confidence interval [CI], 90-99) and 83% (95% CI, 75-93), respectively. In the multivariate Cox model after adjusting for age and gender, patients with higher WB M-MDSC and UnC-MDSC had shorter OS (optimal cutoff points by recursive partitioning analysis, hazard ratio = 7.5 [95% CI, 2.5-22.8], P = .0004; hazard ratio = 3.4 [95% CI, 1.0-11.0], P = .046, respectively).ConclusionIn patients with nonmetastatic UC of bladder, higher WB M-MDSC and UnC-MDSC before cystectomy had negative prognostic value. Prospective validation is warranted.     

Mammary tumorigenesis in chemical carcinogen-treated mice. VII. Prolactin and progesterone levels in BALB/c mice.

Pituitary and serum levels of prolactin (PRL) and serum levels of progesterone (P) were determined by polyacrylamide gel electrophoresis and radioimmunoassays in BALB/c female mice, 15-17 or 44 weeks old, treated with chemical carcinogens. Neither 1.5 mg 3-methylcholanthrene (MCA) nor 1.5-6 mg 7,12-dimethylbenz(a)anthracene (DMBA) markedly altered pituitary or serum levels of PRL in the younger mice, though DMBA increased the total pituitary content of PRL by about 33% in the 44-week-old mice. However, this increase was not correlated with the incidence of mammary tumors in the group or individuals. MCA increased serum P levels by about 22% within 50 days of the last treatment. This increase was attributable to higher serum levels of P during the diestrous and proestrous phases of the cycle. Adrenalectomy reduced serum P levels by about 60%, wheras ovariectomy had no effect. Serum P levels in 44-week-old rats were not affected by DMBA. The results fail to support the notion that MCA and DMBA promote murine mammary tumorigenesis by increasing pituitary and serum prolactin concentrations.     

BALB／c小鼠重度骨髓型急性放射病模型的建立

 目的　建立BALB／c小鼠重度骨髓型急性放射病模型,为重度骨髓型急性放射病的实验研究提供依据。方法　BALB／c小鼠给予60Coγ射线6.0 Gy一次全身照射。观察小鼠照射后一般临床表现、外周血细胞计数、股骨病理组织学及骨髓细胞集落生成情况。结果　小鼠照射后第3天活动量即有不同程度的减少,但均无呕吐、稀便,照射后第11天白细胞降至最低值（基础水平的3.0 %）,照射后第28天恢复至基础水平的53.7 %;照射后第14天血小板降至最低值（基础水平的8.1 %）,照射后第28天恢复至基础水平的60.4 %;照射后第14天骨髓病理示骨髓腔呈空虚状态,骨髓有核细胞集落培养亦提示粒细胞-巨噬细胞集落形成单位（CFU-GM）、混合集落形成单位（CFU-Mix）明显下降;照射后第28天骨髓病理、CFU-GM及CFU-Mix未完全恢复;照射后2个月全部小鼠存活。结论　6.0 Gy 60Coγ射线一次全身照射BALB／c小鼠可成功构建重度骨髓型急性放射病模型,可用于骨髓型急性放射病的实验研究。     

三氧化二砷诱导人鼻咽低分化鳞癌BALB/C裸鼠移植瘤的细胞分化和凋亡的研究(英文)

目的探讨三氧化二砷(As_2O_3)对人鼻咽低分化鳞癌可移植瘤在 BALB/C 裸鼠体内生长的抑制作用并探讨其机制,重点观察其对鼻咽癌细胞的分化诱导作用。方法 CSNE-1鼻咽癌细胞株接种于裸鼠体内构建移植瘤模型。腹腔注入 As_2O_3(5 mg/kg)。光镜及电镜观察移植瘤的形态学变化,TUNEL 染色计算凋亡率,应用免疫组化法检测 PCNA,p53,Bcl-2和 bax 基因的表达。结果腹腔注射 As_2O_3后,鼻咽癌 BALB/C 裸鼠移植瘤生长抑制,瘤组织中癌细胞密度减少,细胞皱缩,胞浆红染,瘤组织分化渐成熟,出现角化细胞和角化珠;间质结缔组织增多。透射电镜下肿瘤细胞出现成熟分化及明显角质化,细胞表面微小突起增多,细胞间桥粒增多并以桥粒互相连接,细胞核浆比例减少,胞浆中出现大量张力原纤维并围绕核周。TUNEL 检查提示凋亡细胞增多;用药后野生型 p53及 bax 高表达,PCNA 低表达,Bcl-2无变化。结论 As_2O_3能诱导人低分化鼻咽裸鼠移植瘤分化和凋亡,可能与 p53及 bax 高表达相关,与 Bcl-2无关。     

三氧化二砷诱导人鼻咽低分化鳞癌BALB/C裸鼠移植瘤的细胞分化和凋亡的研究

目的 探讨三氧化二砷(As2O3)对人鼻咽低分化鳞癌可移植瘤在BALB/C裸鼠体内生长的抑制作用并探讨其机制,重点观察其对鼻咽癌细胞的分化诱导作用。方法 CSNE-1鼻咽癌细胞株接种于裸鼠体内构建移植瘤模型。腹腔注入As2O3(5 mg/kg)。光镜及电镜观察移植瘤的形态学变化,TUNEL染色计算凋亡率,应用免疫组化法检测PCNA,p53,Bcl-2和bax基因的表达。结果 腹腔注射As2O3后,鼻咽癌BALB/C裸鼠移植瘤生长抑制,瘤组织中癌细胞密度减少,细胞皱缩,胞浆红染,瘤组织分化渐成熟,出现角化细胞和角化珠;间质结缔组织增多。透射电镜下肿瘤细胞出现成熟分化及明显角质化,细胞表面微小突起增多,细胞间桥粒增多并以桥粒互相连接,细胞核浆比例减少,胞浆中出现大量张力原纤维并围绕核周。TUNEL检查提示凋亡细胞增多;用药 后野生型p53及baX高表达,PCNA低表达,Bcl-2无变化。 结论As2O3能诱导人低分化鼻咽裸鼠移植瘤分化和凋亡,可能与p53及bax高表达相关,与Bcl-2无关。     

Induction of Adenocarcinomas in the Glandular Stomach of BALB/c Mice Treated with N-Methyl-N-nitrosourea

Male 6-week-old BALB/c strain animals (groups 1 and 2) received 10 weekly intragastric intubations of 0.5 mg/mouse of N-methyl-N-nitrosourea. At week 11 the forestomachs were resected in group 1 but not group 2. Although many animals in group 2 died due to development of squamous cell carcinomas in the forestomach, development of cancers in the glandular stomach was quite similar in both groups. Well-differentiated adenocarcinomas in groups 1 and 2 were found at low incidence at week 20, rising to 100% at week 40, with two lesions metastasizing to the lymph nodes. Four poorly differentiated adenocarcinomas and 5 signet ring cell carcinomas were also found in 27 glandular stomach tumor-bearing animals.     

Mammary adenocarcinomas induced by medroxyprogesterone acetate: hormone dependence and EGF receptors of BALB/c in vivo sublines

Mammary adenocarcinomas were induced by medroxy-progesterone acetate (MPA) in female BALB/c mice. From 5 primary tumors, 9 different sublines were established by s.c. transplantation into syngeneic female mice; these developed after a long latent period (4-12 months). Each subline was transplanted both into 4 mice treated with 40mg of MPA depot (s.c. contralaterally to the tumor inoculum) and into 4 non-treated mice. Of the 9 sublines, 6 proved to be hormone-dependent (MPA-D) and 3 hormone-independent or autonomous (MPA-I). However, even the autonomous lines, when treated with MPA, showed a slight increase in growth. All MPA-D lines had a high content of ER (20-254 fmoles/mg of protein), PR (63-710), PRL-R (44-74) and low or non-detectable EGF-R. Of the 3 MPA-I sublines that were studied, 2 showed a high content of ER (16-125), PR (27-708), PRL-R (19-70) and EGF-R (29-65) while the other one had a low content of ER (0-36), PR (0-13), no EGF-R and moderate PRL-R (15-52). Spontaneous mammary tumors of BALB/c and C3H origin, which also showed an MPA-I pattern of tumor growth, had high levels of EGF-R. We postulate that MPA has a direct effect on mammary tumor cells in MPA-D lines and that the expression of EGF-R is correlated with an autonomous pattern of growth.     

人胚食管异种移植和诱癌研究

［目的］探讨人胚食管在裸鼠体内的异种移植及苯并在诱癌。［方法］人胚食管移植至探鼠，移植的人胚食管用苯并芘诱癌，观察移植物1－4个月并取材进行光镜、电镜和免疫级化检查。［结果］人胚食管在裸鼠皮下可存活4个月以上。移植的人胚食管用苯并芘诱癌，2个月可出现浸润性鳞状上皮癌。其病变有粘膜上皮坏死脱落、再生、增生、乳头状瘤样增生、不典型增生、原位癌及浸润癌。［结论］苯并芘能诱发人食管上皮癌。实验结果将有助于食管癌病因和发病机理研究。     

Obesity and Obese-related Chronic Low-grade Inflammation in Promotion of Colorectal Cancer Development

Colorectal cancer (CRC) is a worldwide health problem, being the third most commonly detected cancerin males and the second in females. Rising CRC incidence trends are mainly regarded as a part of the rapid‘Westernization’ of life-style and are associated with calorically excessive high-fat/low-fibre diet, consumptionof refined products, lack of physical activity, and obesity. Most recent epidemiological and clinical investigationshave consistently evidenced a significant relationship between obesity-driven inflammation in particular stepsof colorectal cancer development, including initiation, promotion, progression, and metastasis. Inflammation inobesity occurs by several mechanisms. Roles of imbalanced metabolism (MetS), distinct immune cells, cytokines,and other immune mediators have been suggested in the inflammatory processes. Critical mechanisms areaccounted to proinflammatory cytokines (e.g. IL-1, IL-6, IL-8) and tumor necrosis factor-α (TNF-α). Thesemolecules are secreted by macrophages and are considered as major agents in the transition between acute andchronic inflammation and inflammation-related CRC. The second factor promoting the CRC development inobese individuals is altered adipokine concentrations (leptin and adiponectin). The role of leptin and adiponectinin cancer cell proliferation, invasion, and metastasis is attributable to the activation of several signal transductionpathways (JAK/STAT, mitogen-activated protein kinase (MAPK), phosphatidylinositol 3 kinase (PI3K), mTOR,and 5’AMPK signaling pathways) and multiple dysregulation (COX-2 downregulation, mRNA expression).     

Benzyl isothiocyanate inhibits murine WEHI-3 leukemia cells in vitro and promotes phagocytosis in BALB/c mice in vivo

Many evidences have shown that dietary intake of cruciferous vegetables could protect against the risk of various types of malignancies. Benzyl isothiocyanate (BITC), one of the compounds from cruciferous vegetables, had shown induced cell cycle arrest and apoptosis in cancer cells. However, there is no available information to address that BITC affects murine leukemia cells in vitro and in vivo. Here, we investigated in vitro effects of BITC on murine leukemia WEHI-3 cells. BITC decreased the percentage of viable cells via G0/G1 arrest and apoptosis in WEHI-3 cells. BITC induced apoptosis through the dysfunction of mitochondria (decreased the levels of mitochondria membrane potential) and activation of caspase-3. Then we investigated in vivo effects of BITC on murine leukemia WEHI-3 cells and the results indicated that BITC decreased the weights of liver and spleen and it also decreased the percentage of CD11b and Mac-3 markers, indicating that the differentiation of the precursor of macrophage and B cells was inhibited. BITC promoted the activity of macrophage phagocytosis in cells which are isolated from PBMC and peritoneal (i.p.). Taken together, BITC can affect WEHI-3 cells in vitro and in vivo.     

Dose-dependent Promoting Effects of Catechol on Glandular Stomach Carcinogenesis in BALB/c Mice Initiated with N-Methyl-N-nitrosourea

The effects of cateehol administration in the diet on stomach carcinogenesis in mice after initiation with N -methyl- N -nitrosourea (MNU) in the drinking water were investigated in a development trial for a new experimental protocol. Male 6-week-old BALB/c mice were given MNU in the drinking water intermittently for a total of three one-week periods, with one-week intervals, at the concentration of 120 ppm (groups 1 and 2). Groups 3 and 4 served as non initiated controls. From week 7, groups 1 and 3 were divided into three subgroups and the mice were fed on diet containing 0.05% (groups la and 3a), 0.2% (groups Ib and 3b), 0.8% (groups 1c and 3c) or 0% (groups 2 and 4) cateehol for 29 weeks. At week 20, appreciably enhanced development of pepsinogen 1-altered pyloric glands was noted in all catechol-treated groups, in a partially dose-dependent manner (12.8±12.5, 13.8±11.7, and 24.0±12.7/100 pyloric glands respectively, for groups 1, 2 and 3). The incidences of adenomas (groups 1, 2 and 3) were also increased. At week 35, dose-dependent induction of adenocarcinomas in groups 1 (3/19), 2 (3/19) and 3 (14/20) was evident. In addition, the depth of invasion of the adenocarcinomas was enhanced by cateehol in a dose-dependent manner, though the histological type was not influenced. Thus, the administration of cateehol in the diet strongly enhanced the pre-neoplastic and neoplastic lesions in mouse glandular stomach induced by MNU in the drinking water, in a dose-dependent manner