Ki67反义多肽核酸抑制肾癌生长的体外及动物实验研究

为研究肿瘤增殖基因Ki67反义多肽核酸(AS-PNAs)对人肾癌细胞的体内外抑制作用.将AS-PNAs(10.0μmol/L)转染人肾癌786-0细胞,采用免疫组化、Western印迹技术检测Ki67表达;3H-TdR掺入试验检测细胞增殖;免疫组化TUNEL法检测细胞凋亡。裸鼠移植瘤内注射AS-PNAs(10.0μmol/L)连续4d后,第3、6、12天处死小鼠,取瘤组织检测肿瘤体积、Ki67表达、细胞凋亡。结果发现,AS-PNAs处理组786-0细胞Ki67表达明显降低,3H-TdR掺入率明显降低,细胞凋亡率明显增加,与随机PNAs对照组比较差异均有显著性(P<0.01)。动物实验AS-PNAs处理组小鼠肿瘤体积明显缩小,Ki-67抗原表达明显降低,细胞凋亡明显增加,与对照组比较差异均有显著性(P<0.01)。Ki67基囚AS-PNAs在体外及动物体内均有抑制增殖、促进凋亡作用,是一种有前途的反义治疗药物。     

死亡肿瘤细胞诱导分化的巨噬细胞体内抗瘤作用的研究

目的观察死亡肿瘤细胞诱导分化的巨噬细胞的体内抗瘤作用。方法采用死亡H22肿瘤细胞诱导单核-巨噬细胞的分化。取40只健康纯系的昆明小鼠，随机分为4组：①生理盐水接种组（A组）；②热灭活H22肿瘤细胞接种组（B组）；③液体石蜡诱生的巨噬细胞接种组（C组）；④死亡肿瘤细胞诱导分化的巨噬细胞接种组（D组）。A组、B组、C组和D组小鼠分别于左后肢皮下接种生理盐水、热灭活H22肿瘤细胞、液体石蜡诱生的巨噬细胞和死亡肿瘤细胞诱导分化的巨噬细胞，2周后各组小鼠分别于左前肢皮下接种生长良好的H22肿瘤细胞，6周后处死全部小鼠，并计算成瘤率、瘤重量与瘤体积；制备小鼠全脾细胞作为效应细胞，以H22肿瘤细胞为靶细胞，混合培养72h后MTT法检测肿瘤细胞杀伤率。结果D组的成瘤率、瘤重量与瘤体积分别为10％、0．12g和0．10cm^3，明显低于A组[80％、（2．41±0．72）g和（2．45±0．82）cm^3]、B组[40％、（0．51±0．36）g和（0．65±0．31）cm^3]和C组[60％、（1．87±0．67）g和（2．15±0．45）cm^3]，P〈0．05，差异有统计学意义；D组的肿瘤细胞杀伤率为（35．89±13．86）％，明显高于A组（12．10±2．74）％、B组（14．88±3．96）％和C组（24．54±4．18）％，P〈0．05，差异有统计学意义。结论死亡肿瘤细胞诱导分化的巨噬细胞具有明显的抗瘤作用。     

Expression of bcl-2 oncoprotein in renal cell tumours

Expression of bcl-2 is associated with inhibition of apoptosis and extension of cell survival. The importance of apoptosis in relation to the development and progression of renal cell neoplasia remains undefined so far. In order to determine the expression of bcl-2 oncoprotein in normal and neoplastic renal cells, 37 renal tumours were investigated by immunolabelling, including 13 clear cell carcinomas, ten tubulopapillary carcinomas, four chromophobic renal cell carcinomas, and ten oncocytomas. Twenty-six samples of adjacent normal renal tissue served as controls. bcl-2 expression was correlated with cell proliferation activity as estimated by Ki67 antigen expression, and p53 protein expression in the tumour samples. The results demonstrate that in the normal kidney, positive bcl-2 immunostaining was present in glomerular parietal epithelial cells, in distal tubular cells, and in sparse proximal tubule cells. Renal cell tumours showed heterogeneous bcl-2 expression according to the tumour cell type. While the majority of carcinomas of clear cell type were usually negative or contained sparsely distributed positive cells, all tubulopapillary carcinomas were consistently positive for bcl-2. In oncocytomas and chromophobic carcinomas, there was a low percentage of bcl-2 immunoreactive tumour cells; some nuclear bcl-2 positivity was detected in one chromophobic tumour. These findings indicate variable bcl-2 oncoprotein expression in different types of renal cell tumours, with the highest level of expression in tubulopapillary carcinomas. No clear relationship was found between nuclear grade, cell proliferation activity, and level of bcl-2 expression. p53 protein was detected in only one tubulopapillary carcinoma.     

Tertiary lymphoid structures generate and propagate anti-tumor antibody-producing plasma cells in renal cell cancer

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The F‐prostaglandin receptor is a novel marker for tumor endothelial cells in renal cell carcinoma

Tumor angiogenesis is necessary for tumor progression and metastasis; therefore, tumor blood vessels are potential therapeutic targets in anticancer therapy. We previously reported that tumor endothelial cells (TECs) exhibit different phenotypes compared with normal endothelial cells (NECs), and microarray analyses of mouse TECs and NECs have shown that several genes are upregulated in TECs compared with NECs. Among these genes, the expression levels of prostaglandin F receptor (PTGFR) mRNA, which encodes the prostaglandin F receptor (FP), were higher in TECs than in NECs. It has been reported that FP and its ligand, prostaglandin F_2α, are involved in tumor angiogenesis. However, there have been no reports of the expression of PTGFR in the tumor vessels of renal cell carcinoma (RCC). Thus, we isolated human TECs (hTECs) from RCCs. The expression levels of PTGFR mRNA were also upregulated in hTECs. In addition, immunostaining showed that the PTGFR was expressed in human tumor blood vessels in vivo. These findings suggested that PTGFR is a novel TEC marker and that it may be a novel target for antiangiogenic therapy for RCC.     

Effect of exogenous poly(A)+mRNA on antigenic properties and growth of Krebs-2 tumor cells

Laboratory of Molecular Genetic Bases of Oncogenesis, Institute of Cytology and Genetics, Siberian Branch, Academy of Sciences of the USSR. Laboratory of Structure and Function of the Gene, Institute of Bioorganic Chemistry, Siberian Branch, Academy of Sciences of the USSR, Novosibirsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. P. Kaznacheev.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 113, No. 3, pp. 301–303, March, 1992.     

十溴联苯醚对宫颈癌细胞体外增殖的影响

目的探讨十溴联苯醚（PBDE-209）对宫颈癌HeLa细胞体外增殖的影响。方法观察HeLa细胞经不同浓度PBDE-209暴露后发生的形态学变化,并用MTT方法检测细胞存活率改变,采用免疫荧光化学方法检测细胞增殖核抗原Ki67表达变化,流式细胞术观察HeLa细胞细胞周期的分布变化。结果随着浓度升高,细胞增殖明显,细胞间隙变小。MTT方法检测发现PBDE-209促进HeLa细胞增殖,细胞增殖率的升高呈时间和浓度依赖效应,200nmol/LPBDE-209作用24、48和72h后细胞增殖率分别是阴性对照组的1.2、1.5和1.8倍（P〈0.05）。Ki67表达强度随着PBDE-209浓度升高而增强。流式细胞术检测发现200nmol/LPBDE-209作用72h后,G0/G1期细胞比例下降了8%,而S期细胞比例升高了7%,与阴性对照组比较差异均有统计学意义（P〈0.05）。结论低浓度PBDE-209暴露使HeLa细胞G0/G1期细胞减少,S期细胞增加,从而促进细胞增殖。     

膜联蛋白A7低表达对人肝癌HepG2细胞增殖的影响

目的 探讨膜联蛋白A7低表达对人肝癌HepG2细胞的增殖产生的影响。方法 采用Western blotting法鉴定siRNA可有效抑制膜联蛋白A7表达,然后用脂质体转染法将siRNA转染入HepG2细胞,将细胞分为siRNA干扰组、阴性对照组和空白对照组,在转染后24h、48h、72h进行细胞计数以绘制细胞增殖曲线,转染后48h进行MTT实验以检测细胞增殖活力;免疫组织化学法检测cyclinD1和Ki67的表达,Western blotting法检测cyclinD1的表达情况。结果 转染了靶向膜联蛋白A7的siRNA后48h的HepG2细胞,细胞计数和MTT实验可见siRNA干扰组细胞增殖活力较阴性对照组和空白对照组显著降低（P<0.05）;cyclinD1和Ki67蛋白表达均为siRNA干扰组细胞表达显著低于两对照组（P<0.05）。 结论 膜联蛋白A7低表达可能对HepG2细胞的增殖具有一定的抑制作用。     

The increasingly anti-tumor effect of a colonic carcinoma DNA vaccine carrying HER2 by the adjuvanticity of IL-12

The present study aimed to determine the effect of recombinant DNA vaccine-based human epidermal growth factor receptor-2 (HER2) and Interleukin 12 (IL-12) on the development of colonic carcinoma in mice and the potential immune mechanisms involved. Recombinant plasmids pVAX1-HER2, pVAX1-IL-12 and pVAX1-HER2-IL-12 were constructed, and injected into female mice intramuscularly (i.m.) followed by an electric pulse. The humoral and cellular immune responses after immunization were examined by enzyme linked immunosorbent assay (ELISA) and enzyme-linked immunospot assay (ELISPOT), respectively. To evaluate the anti-tumor efficacy of the plasmids, a mouse model with a HER2-expressing tumor was designed. Mice vaccinated with the HER2-IL-12 plasmid generated the strongest inhibition efficacy on the growth of HER2-expressing tumors and prolonged mouse survival. These observations emphasized the potential of IL-12 as an adjuvant for DNA vaccines and of vaccines based on HER2 and IL-12 as a promising treatment for colonic carcinoma.     

Expression of annexins on the surfaces of non-metastatic and metastatic human and rodent tumor cells

Annexins are a large group of calcium-dependent cytoskeletal- and membrane-associated proteins whose properties include cytoskeleton and phospholipid binding and mitotic signal transduction. Although annexin-like molecules have been reported on the external plasma membranes of certain cells, in general they are considered to be cytoplasmic proteins. We report here the heterogenous expression of certain annexins (I-VI) on the external cell surfaces of non-metastatic and metastatic murine (RAW117 large-cell lymphoma), rat (13762NF mammary adenocarcinoma) and some human (KM12 and HT29 colorectal carcinoma) cell lines but not on some other cell lines such as human (A375 and McWo) and mouse (B16) melanoma. The implication of annexin cell surface expression in the metastatic process is discussed with respect to tumor cell adhesion.     

LMP2, a novel immunohistochemical marker to distinguish renal oncocytoma from the eosinophilic variant of chromophobe renal cell carcinoma

LMP2 is a subunit of the immunoproteasome that is overexpressed in oncocytic lesions of the thyroid gland. This study was designed to assess the expression profile and diagnostic utility of LMP2 in two renal oncocytic tumors that share similar morphologic features but have different clinical outcomes: renal oncocytoma (RO) and the eosinophilic variant of chromophobe renal cell carcinoma (CHRCC-EO). A total of 56 RO, 38 classic CHRCC, and 7 CHRCC-EO cases, as well 84 normal kidney controls, were selected from the Johns Hopkins surgical pathology archive and stained for LMP2 using a standard immunohistochemical protocol. Sections were scored for cellular location (nuclear versus cytosolic), intensity (from 0 to 3), and percent of area involved (from 0 to 100%), and an H score was calculated multiplying the intensity by the extent of the staining signal. The cytoplasmic expression of LMP2 was similar among the renal lesions, being present in 44 of 56 (79%) ROs, 27 of 38 (71%) CHRCCs, and 7 of 7 (100%) CHRCC-EO cases. The nuclear expression of LMP2, however, was more informative. All CHRCC-EO cases (7 of 7, 100%) strongly showed nuclear LMP2 staining, as opposed to only 2 of 56 (4%, P < 0.0001) ROs and 9 of 38 (24%, P = 0.0001) classic CHRCCs. These results suggest that the nuclear LMP2 expression can be used in clinical scenarios where histological distinction between RO and CHRCC-EO remains challenging.     

碘佛醇对兔VX2肾癌超声血流显像影响的研究

目的评价碘佛醇在兔VX2肾癌超声血流显像中的作用。方法24只实验兔建立肾VX2肿瘤模型，随机分3批，每批8只，分别于种植后第7、14、21天，静脉注射碘佛醇，观察注射前后肿瘤彩色血流信号的变化情况。结果注射碘佛醇后肿瘤内彩色血流信号较注射前增多、增强。其中第7天，肿瘤的CDFl分级变化无显著统计学意义，第14天和第21天差异有显著统计学意义。注射碘佛醇后连续观察20min，血流信号增强仍保持在高峰状态。结论碘佛醇对兔VX2肾癌的彩色多普勒血流显像起增强作用。     

A case report of brown tumor in a patient with chronic renal failure and renal cell carcinoma

We report a case of a 72 year old male with hyperparathyroidism secondary to end stage diabetic renal disease and coexisting bilateral chromophobe renal cell carcinomas. The patient presented with back and groin pain, right pelvic hemorrhage, and multiple lytic bone lesions concerning for metastatic renal cell carcinoma. Fine needle aspiration cytology demonstrated benign appearing osteoclasts and spindled cells. A concurrent core biopsy showed foci of spindled cell proliferation populated by osteoclast‐like giant cells with stromal hemorrhage without evidence of metastatic carcinoma. The cytologic and histologic findings, in correlation with the clinical history, radiographic features, markedly increased parathyroid hormone levels and other serologic studies, were diagnostic of the reactive lesion seen in brown tumor of hyperparathyroidism secondary to chronic renal failure.     

蝎毒抗癌多肽对大肠癌细胞LoVo的影响及其与化疗药物的协同作用

目的: 探讨东亚钳蝎蝎毒抗癌多肽对大肠癌细胞LoVo的影响及其与多种化疗药物的联合作用。方法: 采用MTT法, 测定蝎毒抗癌多肽与各种化疗药物对LoVo细胞的增殖抑制作用以及蝎毒抗癌多肽与化疗药物联合对LoVo细胞的增殖抑制作用。结果: 蝎毒抗癌多肽可抑制LoVo细胞增殖, 并可诱导其凋亡。低浓度的蝎毒抗癌多肽与化疗药物联合用药, 可产生较强的抑制细胞增殖的作用。结论: 蝎毒抗癌多肽具有直接的抗肿瘤作用, 并且具有可以增强化疗药物作用的功能。     

舒尼替尼对转移性肾癌患者髓系来源抑制细胞的影响

舒尼替尼是一种口服的多靶点酪氨酸激酶抑制剂,具有抗血管生成和抑制肿瘤增殖的作用.与IFN-α相比,舒尼替尼明显提高了转移性肾癌(mRCC)患者的无进展生存期,并成为治疗mRGC的一线治疗方案.近期有研究报道,舒尼替尼还可以通过抑制髓系来源抑制细胞(MDSC)调节肿瘤免疫.MDSC是一群来源于髓系细胞谱系的异质细胞,其通...     

肿瘤浸润淋巴细胞与IL-2联合诱导Lewis肺癌凋亡的研究

目的：研究肿瘤浸润淋巴细胞（ＴＩＬ）与ＩＬ－２联合体内诱导凋亡产生抗瘤作用的机理。为应用ＴＩＬ治疗肺癌提供依据。方法：从Ｌｗｉｓ肺癌（ＬＬＣ）瘤体中分离提了参含１０００Ｕ／ｍｌＩＬ－２的完全培养基中培养后，与ＬＬＣ细胞中１０：１接种于Ｃ５７ＢＬ／６小鼠腋下，每日１０００Ｕ／ｍｌＩＬ－２瘤内注射，并测量肿瘤体积，于接种后第１８天处死小鼠。结果：发现ＴＩＬ加ＩＬ－２有明显地抑制ＬＬＣ生长的作用，ＨＥ感     

Expression of EphA2 protein is positively associated with age,tumor size and Fuhrman nuclear grade in clear cell renal cell carcinomas

The receptor tyrosine kinase of EphA2 has been shown frequently overexpressed in various types of human carcinomas, which implicated that it plays important roles in carcinogenesis. Although EphA2 protein expression has been investigated in many types of human carcinomas, the relationship between the expression of EphA2 protein in clear cell renal cell carcinoma was not well documented. In the present study, using specific anit-EphA2 polyclonal antibody and immunohistochemistry, we evaluated EphA2 protein expression levels in clear cell RCC specimens surgically resected from 90 patients. Our results shows that EphA2 protein was positively expressed in all normal renal tubes of 90 samples (100%, 3+), which was expressed at low levels in renal cortex but high levels in the collecting ducts of the renal medulla and papilla. EphA2 was negatively or weakly expressed in 30 out of 90 samples (33.3%, 0/1+), moderately expressed in 24 samples (26.7%, 2+) and strongly expressed in 36 samples (40%, 3+). Expression of EphA2 was positively associated with age (P=0.029), tumor diameters (P<0.001) and Fuhrman nuclear grade (P<0.001). Our results indicate that EphA2 variably expressed in clear cell renal cell carcinomas. High expression of EphA2 was more often found in big size and high nuclear grade tumors, which indicated EphA2 protein may be used as a new marker for the prognosis of clear cell renal cell carcinoma.     

COX-2参与戊地昔抑制Lewis肿瘤生长

目的探讨戊地昔布对Lew is肿瘤的抑制作用及与抑制环氧化酶-2(COX-2)是否有关。方法用W estern b lot检测COX-2的表达,用ELISA检测前列腺素E2(PGE2),用HE染色检测肿瘤组织淋巴细胞的浸润。结果戊地昔布可抑制肿瘤生长,提高荷瘤小鼠的生存率;戊地昔布对COX-2表达没有明显影响,但降低肿瘤部位PGE2的含量;戊地昔布增加肿瘤组织淋巴细胞浸润,不影响荷瘤小鼠胃肠道上皮细胞结构以及出、凝血时间。结论戊地昔布可抑制Lew is肿瘤的生长,与抑制肿瘤组织COX-2的活性,降低PGE2含量有关。     

建立兔胫骨VX2骨肿瘤模型的方法

文题释义：VX2 肿瘤：是一种可移植的恶性乳头状瘤,可移植在兔的肝脏、肺脏、骨骼肌肌肉等处,所建立的肿瘤模型具有较高的生物学特性。穿刺活检：是骨与软组织肿瘤获取组织病理诊断的主要方法,经肿瘤体表投影或影像学引导下使用穿刺设备进行瘤体组织的获取。背景：关于兔的VX2肿瘤模型方法有多种,但各方法的可靠性无人探究,课题组在原有方法上进行了改良,进行了此项研究。目的：探讨兔VX2肿瘤组织块、细胞悬液分别经改良法与传统植入法进行兔胫骨VX2骨肿瘤造模的可靠程度。方法：将健康成年新西兰大白兔40只随机分为2组,其中组织块组植入肿瘤组织块进行胫骨VX2肿瘤造模,细胞悬液组植入肿瘤细胞悬液进行胫骨VX2肿瘤造模,2组实验动物左侧胫骨均采用穿刺针植入,右侧胫骨均采用传统法植入。造模成功1 h后对穿刺孔周围进行超声检查明确穿刺孔有无血肿,于第3周处死所有实验动物,进行双侧胫骨X射线检查明确肿瘤生长范围,取肿瘤及穿刺部位周围软组织进行大体及病理学检查对比肿瘤大小,并明确有无肿瘤细胞局部浸润。结果与结论：①组织块组有1只实验动物于术后死亡,脱组,细胞悬液组所有实验动物均存活;②X射线检查可见组织块组肿瘤均侵犯皮质层,细胞悬液组皮质层未侵及;肿瘤大体观察可见组织块组肿瘤侵犯骨皮质长度大于细胞悬液组;③组织块组改良法、传统法植入1 h后分别有1例,7例穿刺孔周围血肿;细胞悬液组改良法、传统法植入1 h后分别有2例,9例穿刺孔周围血肿;④组织块组改良法、传统法植入后3个月分别有1例,8例出现肿瘤局部浸润;细胞悬液组改良法、传统法植入后分别有2例,11例出现肿瘤局部浸润;⑤结果表明,组织块植入较细胞悬液植入制作兔VX2骨肿瘤更易培养出大的瘤体,从而使实验更易于观察,且组织块植入后肿瘤局部浸润率较细胞悬液法要低;同时采用改良法植入可有效降低造模过程中肿瘤细胞局部浸润率。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程