Biologically active sphingolipids in transplantable tumors of different histogenesis

The composition of biologically active sphingolipids in hepatoma 22, breast adenocarcinoma, Lewis lung carcinoma, large intestine adenocarcinoma, cervical carcinoma, and melanomas M3 and B16 was compared to elucidate the similarity and differences in sphingolipids of subcutaneously transplantable murine tumors. The sphingolipid composition of the tumors was found to widely vary. The sphingomyelin, ceramide, glucosyl-and lactosylceramide, and ganglioside GD3 contents in hepatoma 22 are higher than those in normal tissue. No common regularities for tumors of different origin were observed in the ratios of bioeffectors inhibiting proliferation and stimulating apoptosis and bioeffectors stimulating cell growth and survival. However, the Cer/(GlcCer + LacCer) ratios were very low and practically equal in two melanoma strains, which probably indicates the degree of tumor malignancy. The results suggest that the content and composition of sphingolipids in tumors depend on their histogenesis.     

The changes in the sphingenine/sphinganine ratio in sphingolipids of transplantable rat tumors depends on a transplantation organ

The content of sphingenine (sphingosine) and sphinganine was determined in the total pool of sphingomyelin and ceramide in the rat tumors transplanted subcutaneously and intrahepatically. The sphingenine/sphinganine ratio in the subcutaneously transplanted sarcoma M1 and cholangiocellular carcinoma RS1 was lower than that in the sphingolipids of the intrahepatically transplanted tumors. However, the sphingenine/sphinganine ratio in the subcutaneously transplanted rat hepatoma 27 was higher than in the intrahepatically transplanted hepatoma. These observations indicate that the sphingenine/sphinganine ratio in sphingolipids of tumors depends on the tumor type and its cellular microenvironment.     

The Sphingenine/Sphinganine Ratio in Sphingolipids of Transplantable Rat Tumors Depends on a Transplantation Organ

The content of sphingenine (sphingosine) and sphinganine was determined in the total pool of sphingomyelin and ceramide in the rat tumors transplanted subcutaneously and intrahepatically. The sphingenine/sphinganine ratio in the subcutaneously transplanted sarcoma M1 and cholangiocellular carcinoma RS1 was lower than that in the sphingolipids of the intrahepatically transplanted tumors. However, the sphingenine/sphinganine ratio in the subcutaneously transplanted rat hepatoma 27 was higher than in the intrahepatically transplanted hepatoma. These observations indicate that the sphingenine/sphinganine ratio in sphingolipids of tumors depends on the tumor type and its cellular microenvironment.     

Clinicopathological study of involvement of granulocyte colony stimulating factor and granulocyte-macrophage colony stimulating factor in non-lymphohematopoietic malignant tumors accompanied by leukocytosis

Involvement of granulocyte colony stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF) in non-lymphohematopoietic malignant tumors accompanied by leukocytosis was clinicopathologically investigated. Among 1,778 autopsy cases in the last 20 years, 485 lesions of 439 cases with non-lymphohematopoietic malignant tumors accompanied by leukocytosis with a white blood cell count of 10,000/mm3 or greater during the course were immunohistologically examined for G-CSF and GM-CSF. Three (0.7%) and two cases (0.5%) were G-CSF- and GM-CSF-positive, respectively. GM-CSF mRNA was confirmed by using non-fixed cryopreserved tumor tissues in one case positive for GM-CSF. G-CSF-positive cases were large cell carcinoma of the lung, adenocarcinoma of the colon, and adenocarcinoma of the stomach, and GM-CSF-positive cases were spindle cell carcinoma of the lung and malignant thymoma. In the case with stomach carcinoma, the primary lesion showing moderately differentiated adenocarcinoma was negative, but the lung metastatic lesion showing less differentiated adenocarcinoma was G-CSF-positive. The survival period was six months or less in four out of five positive cases. The highest white blood cell count in five CSF-positive cases was markedly elevated: 29,400-103,500/mm3 (mean: 59,700/mm3). In four cases, excluding one case which may have been markedly affected by chemotherapy, the bone marrow showed hyperplasia, and the number of the granulocyte series cells significantly increased. There were three cases (0.7%) negative for both G-CSF and GM-CSF, although they showed marked leukocytosis (60,000/mm3 or higher) which were higher than the mean count of CSF-positive cases and was not observed in autopsy cases with non-tumorous diseases. Other stimulating factors may be involved in the development of leukocytosis in such cases.     

Detection of human papillomavirus in non-small cell lung carcinoma by polymerase chain reaction

Human papillomavirus (HPV) infection is one of the risk factors contributing to the pathogenesis of lung cancer. The aim of the study was to determine the presence of HPV in non-small cell carcinomas of the lung. The study included 40 tumors: 22 squamous cell carcinomas, 13 adenocarcinomas and 5 large cell carcinomas. HPV was found in 4 cases (10%). High risk HPV was present in 3 tumors: in one squamous cell carcinoma, one large cell carcinoma and one adenocarcinoma, while low risk HPV was detected in one adenocarcinoma.     

裸鼠肿瘤动物模型VEGF受体表达及其意义

目的　通过免疫组织化学染色了解flt 1与flk 1 KDR(VEGF的两个高亲和受体 )在人肿瘤细胞皮下接种肿瘤动物模型的血管内皮细胞与肿瘤细胞中的表达。方法　取荷瘤裸鼠皮下接种瘤块 ,漂洗、固定、石蜡连续切片 ,进行两种受体相应免疫组化检测。结果　在 13种荷瘤裸鼠血管内皮细胞及肿瘤细胞中flt 1的阳性率大部分为强阳性或中阳性 ,而只有在荷人胃腺癌MKN 4 5裸鼠的肿瘤细胞中flt 1的阳性率为弱阳性 ,在荷人卵巢癌SKOv3裸鼠的肿瘤细胞中flt 1的表达为阴性。相比较而言 ,在 13种荷瘤裸鼠血管内皮细胞及肿瘤细胞中KDR的阳性率大部分为中阳性或弱阳性 ,并且在荷人肝癌SMMC 772 1裸鼠 ,荷人胃腺癌SPC A1裸鼠 ,荷人高转移肝癌移植瘤裸鼠 ,荷人卵巢癌SKOv3裸鼠的肿瘤细胞中 ,荷人宫颈癌移植瘤裸鼠和荷人胃腺癌MKN 4 5裸鼠的肿瘤细胞中 ,KDR表达为阴性。结论　VEGF受体共同表达于肿瘤血管内皮细胞与肿瘤细胞 ,提示了VEGF与VEGF受体结合作用在肿瘤演化中的重要性 ,为靶向于VEGF受体的基因治疗策略选择裸鼠动物模型提供了参考依据     

自发性可移植性615小鼠肺癌和肝癌组织神经节苷脂高效薄层层析图谱的分析

本文研究了615近交系小鼠的自发性可移植性肺癌(P_(615))和肝癌(H_(615))组织中神经节苷脂结合唾液酸含量和神经节苷脂图谱。P_(615)和H_(615)癌组织中GLS结合唾液酸含量均比正常对照为高,分别为对照组织的2.9倍和1.9倍。615小鼠的正常肺组织和肝组织的GLS主要成分均为GM_3。在P_(615)和H_(615)癌组织中GM_3含量均明显减少。肺癌组织中GM_2大量增加,肝癌组织中不仅GM_2明显增加,GM_1和GD_(18)也明显增加。P_(615)和H_(615)这两种分化程度较高、恶性程度较低的癌组织GD_3的百分含量比正常对照组织略有降低。本文结果提示,自发性可移植性P_(615)和H_(615)肿瘤组织中不仅神经节苷脂含量(以唾液酸量计)升高,而且GLS的组分也发生改变。GM_3含量减少和GM_2含量增高可能与肿瘤的恶性生长和分化程度有关。     

Squamous cell carcinoma antigen in genitourinary tumors

We have evaluated the serum levels of squamous cell carcinoma (SCC) antigen in patients with adenocarcinoma of the kidney, adenocarcinoma of the prostate, germ cell tumors of the testis, transitional cell carcinoma of the bladder, and SCC of the penis, urethra, and bladder. Serum SCC antigen levels were elevated in 5 of 11 patients (45%) with metastatic SCC of the penis, and in the 3 patients for whom serial determinations were made, the serum levels correlated correctly with the progression of disease or response to treatment. The antigen was elevated in 1 of 3 patients with SCC of the urethra, and 1 apparent false-positive value was observed in a patient with adenocarcinoma of the prostate. Otherwise, no SCC antigen elevations were noted among 10 patients with metastatic adenocarcinoma of the prostate, 8 with metastatic adenocarcinoma of the kidney, 11 with metastatic transitional cell carcinoma of the bladder, 8 with metastatic nonseminomatous germ cell tumors of the testis, and 2 patients with metastatic SCC of the bladder.     

Comparative study of nucleosome particles in chromatin from normal and tumor cells. I. Structural parameters

The composition and structure of nucleosomic fragments isolated from the ascitic hepatoma 22A cells, liver and from cells of C3HA mice in norm and after partial hepatectomy were investigated. Via electrophoresis in 1.5% agarose gel with the emplogment of reperic restrictive DNA fragments and with the help of mathematical processing, the value of the nucleosomic DNA repeat in ascitic hepatoma 22A was calculated to be 187 b.p., and in regenerating liver--196 b.p. The absence of the H1 degree subfraction in chromatin of ascitic hepatoma 22A cells was found. Lower electrophoretic mobility in 5% polyacrylamid gel of nucleosomic chromatin fragments of ascitic hepatoma 22A as compared with their counterparts from healthy mice liver was established. The method of circular dichroism allowed to reveal differences in the RNA and protein structural state in nucleosomes of normal and tumour cells. The structure of nucleosomes of regenerating mice liver of the C3HA strain did not differ from that of normal liver of the same mice.     

Base composition studies on mitochondrial 4 S RNA from rat liver and Morris hepatomas 5123D and 7777.

The major and modified base composition of mitochondrial 4 S RNA from rat liver and from Morris hepatomas 5123D and 7777 has been determined for 16 constituents using a chemical tritium-derivative method. The base composition of these mitochondrial 4 S RNA preparations was compared with the base composition of cytoplasmic and bacterial (Escherichia coli B and Bacillus subtilis) 4-S RNAs. The results of these studies are: 1. When compared with cytoplasmic 4 S RNA, the liver and hepatoma mitochondrial 4-S RNAs are characterized by high (A + U)/(G + C) ratios and low overall degrees of base methylation and modification. 2. The mammalian mitochondrial 4-S RNAs are qualitatively even more different from the bacterial 4-S RNAs than from their cytoplasmic counterparts. Thus, several modified constituents found in both cytoplasmic and mitochondrial 4 S RNA are absent from the bacterial 4-S RNAs. 3. Mitochondrial 4S RNA from both hepatomas was found to be under-methylated and undermodified when compared with normal liver mitochondrial 4S RNA. This trend is more pronounced for the rapidly growing hepatoma 7777 (i.e., 17% undermethylation) than for the more slowly growing hepatoma 5123D (i.e., 8% undermethylation). These findings are discussed in relationship to (1) results of other authors on composition of mitochondrial 4 S RNA, (2) special features of structure and biosynthesis of mitochondrial 4 S RNA, (3) the possible evolutionary origin of mitochondria and (4) the possible role played by aberrant mitochondrial 4 S RNA in altered mitochondrial protein synthesis in tumors.     

Some characteristics of isoenzymic spectrum of hexokinase and glucose levels in hepatomas and liver of the host]

The total activity of hexokinase (HK) and HK isoenzymic spectrum of the normal liver and slowly groming hepatoma 49 did not show any essential differences. However, the HK total activity and the relative and absolute contents of isoenzyme HK-3 were increased in hepatomas 61 and especially in the rapidly growing hepatoma 22-a. The glucokinase activity decreases in the hepatiomas 49 and 61 and disappears in the rapidly growing hepatoma 22-a. The glucose content in hepatoma 49 was slightly lower than in the normal liver, whereas in other hepatoma no traces of glucose could be detected. At low glucose concentration in the medium (0,1 mM), i.e. under conditions simulating those characteristic of tumors in vivo, the predominant form of HK in all hepatomas studied was found to be HK-3. In the liver of hepatoma-bearing mice some shifts in the value of total HK activity and its isoenzymic spectrum, reminding one of those found in hepatomas themselves, were observed. Unequal deviations in the total HK activity and its isoenzymic spectrum in hepatomas with different degrees of malignancy indicate that these characteristics are secondary rather than primary events depending on tumour progression.     

Differences in Lipid Composition and Proliferative Activity of Rat Hepatoma-27 Depending on the Target Organ

Proliferative activity and lipid composition (phospholipids and gangliosides) were studied in rat hepatoma-27 transplanted subcutaneously or intrahepatically (as models for primary and metastasizing tumors). The mitotic index of subcutaneously transplanted hepatoma far exceeded that of the intrahepatically transplanted tumor. The overall amounts of both phospholipids and gangliosides increased appreciably in the subcutaneously growing hepatoma (in contrast to the intrahepatically growing tumor) in comparison to the control hepatic tissue. The ganglioside composition in the tumors differs from that in the liver: ganglioside GD3 appears, whereas gangliosides GD1b and GT1b decrease in amount in the intrahepatic tumor compared to the control liver and disappear in the subcutaneously transplanted hepatoma. In both tumor types, the amounts of both phosphatidylethanolamine and sphingomyelin exceed the control values. Comparison of these results with previously reported data concerning the phospholipid and ganglioside composition in the regenerating rat liver indicates that the difference in the lipid composition between the subcutaneously and intrahepatically growing hepatomas-27 is due to their different proliferative status and also their microenvironment.     

The effect of radiation with polychromatic visible and infrared light on the tumorigenicity of murine hepatoma 22A cells and their sensitivity to lysis by natural killers

The tumorigenicity of murine hepatoma cells (MH-22a) and their sensitivity to lysis by natural killers (NKs) have been studied after exposure to polychromatic visible and infrared light (VIS-IR, 480–3400 nm, 40 mW/cm²), similar to the terrestrial solar spectrum without its minor UV component, with the aim of clarifying the participation of this important environmental and physiotherapeutic factor in regulation of antitumor protective system. MH-22 cells were exposed in vitro to VIS-IR light and their sensitivity to lytic activity of NKs was evaluated. It was found that, after exposure to VIS-IR light at a dose of 4.8 J/cm², the sensitivity of MH-22a cells to lysis by NKs increased by 1.5–2 times, while after exposure at a dose of 9.6 J/cm² it did not change at all the ratios of the NKs-number (effectors) to that of hepatoma cells — targets (1 : 5–1 : 50). An increase of the hepatoma cell sensitivity to NKs was accompanied by structural changes of cell surface: the capability of supramembranous glycoproteins (glycocalyx) to sorb the vital dye alcian blue (AB) was significantly lower than in the case of unexposed cells of the control group. However, no changes in AB sorption was revealed in hepatoma cells exposed to light at a dose of 9.6 J/cm². The tumorigenicity of photoirradiated MH-22a cells has been studied in the experiments in vitro. For 25 days after transplantation of light-exposed hepatoma cells to C3HA syngene mice, the tumor volume proved to be smaller after exposure to light at both doses of 4.8 and 9.6 J/cm² than in the control group (by 4–4.5 times and 2.5–4 times, respectively), which correlated with an increase of sensitivity to lysis by NKs and with a decrease of AB sorption after light exposure only at a dose of 4.8 J/cm². Using the flow-cytometry method, we could show that VIS-IR light at the doses used did not interfere with the distribution of hepatoma cells over the cell-cycle phases and, thus, deceleration of the tumor growth was not associated with a cytostatic effect of the VIS-IR light. To evaluate the effect of polychromatic light on growth of the preformed tumors, a 5-day course of daily light exposure of C3HA tumor-bearing mice was performed on the 10th day after subcutaneous transplantation of 2 × 10⁵ cells of syngene hepatoma, when tumors developed in all (100%) animals. As in the case of transplantation of light-exposed cells, irradiation of tumor-bearing mice at doses 4.8–9.6 J/cm² resulted in a deceleration of tumor growth (by 2.1–2.9 and 2,2 times, respectively) for 4 weeks compared with unirradiated mice.     

Discussion of Paper on “Brilliant-Green-Bile Media”

Abstract

Insulin-like growth factor II-messenger RNA-binding protein 3 (IMP3) is an oncofetal RNA-binding protein that promotes tumor cell proliferation by enhancing IGF-II protein synthesis and inducing cell adhesion and invasion by stabilizing CD44 mRNA. IMP3 expression has been studied in many human neoplasms with growing evidence that IMP3 is a biomarker of enhanced tumor aggressiveness. IMP3 expression has been correlated with a poorer phenotypic profile including increased risk of metastases and decreased survival. Only a few studies have examined IMP3 expression in lung cancers. We review here the literature concerning IMP3 expression in lung neoplasms, specifically adenocarcinoma, squamous cell carcinoma, and neuroendocrine tumors of the lung. IMP3 immunohistochemical expression was reported in 27-55% of cases of primary pulmonary adenocarcinoma and in 75-90% of cases of squamous cell carcinoma of the lung. In adenocarcinoma, IMP3 expression was reported to be correlated with more poorly differentiated histological grade, advanced stage of disease and lymph node metastases. IMP3 expression also may be a marker of high grade pre-invasive squamous lesions including high grade dysplasia and carcinoma in situ. In neuroendocrine tumors of the lung, IMP3 expression was expressed in all reported cases of large cell neuroendocrine carcinoma and small cell lung carcinoma, but expression was limited in carcinoid tumors. Overall, IMP3 appears to be a useful diagnostic marker for lung cancer pathology including for discriminating high grade neuroendocrine tumors and low grade carcinoids and for identifying high grade pre-invasive squamous lesions.     

The stimulation of DNA synthesis in resting Swiss 3T3 cells by non-histone chromosomal proteins

Nonhistone chromosomal proteins (NHC), isolated from the kidney and liver of intact rats, the liver of rats treated with hepatocarcinogen DEN and the rat hepatoma, stimulate DNA synthesis in Swiss 3T3 cells in resting culture. The maximum stimulating effect was obtained in the presence of narrow NHC fractions eluted with 0.4-0.5 M NaCl from the phosphocellulose column and identified as hetero-organic NHC protein antigens of the kidney origin associated with hepatocellular tumors.     

Synergistic action of tiazofurin and difluorodeoxycytidine on differentiation and cytotoxicity.

Tiazofurin (TR), an inhibitor of IMP dehydrogenase, causes remissions and induced differentiation in human leukemia through lowering the concentrations of GTP and dGTP. A deoxycytidine analog, difluorodeoxycytidine (DFDC), is an anti-tumor agent phosphorylated by deoxycytidine kinase, resulting in decreased concentration of dCTP, leading to inhibition of DNA synthesis. In HL-60 cells DFDC induced differentiation and inhibited proliferation in a dose-dependent manner (IC50 = 4 nM); TR provided synergism with DFDC. DFDC inhibited proliferation in OVCAR-5 human ovarian carcinoma cells (IC50 = 25 nM) and colony formation in PANC-1 human pancreatic carcinoma cells (IC50 = 2 nM) and rat hepatoma 3924A cells (IC50 = 22 nM). TR and DFDC are synergistically cytotoxic in hepatoma cells and additive in PANC-1 cells. The two drugs together should be helpful in treating leukemias and solid tumors in humans.     

Change in the resistance of partially hepatectomized mice to transplanted tumors in the period of liver regeneration completion]

There were two inhibition periods of the growth of ascitic hepatoma 22a when its cells were transplanted intraperitoneally to partially hepatectomized A/he mice 1 to 12 days after the operation. The first inhibition took place when the transplantation was performed one day, and the second--3 to 10 days, after partial hepatectomy. The animals which were most resistant to this tumour (5 to 8 days after the operation) proved to be resistant, although to a lesser extent, to the intraperitoneal transplantation of sarcoma 37 and Ehrlich's adenocarcinoma.     

Immune and Inflammatory Cell Composition of Human Lung Cancer Stroma

Recent studies indicate that the abnormal microenvironment of tumors may play a critical role in carcinogenesis, including lung cancer. We comprehensively assessed the number of stromal cells, especially immune/inflammatory cells, in lung cancer and evaluated their infiltration in cancers of different stages, types and metastatic characteristics potential. Immunohistochemical analysis of lung cancer tissue arrays containing normal and lung cancer sections was performed. This analysis was combined with cyto-/histomorphological assessment and quantification of cells to classify/subclassify tumors accurately and to perform a high throughput analysis of stromal cell composition in different types of lung cancer. In human lung cancer sections we observed a significant elevation/infiltration of total-T lymphocytes (CD3⁺), cytotoxic-T cells (CD8⁺), T-helper cells (CD4⁺), B cells (CD20⁺), macrophages (CD68⁺), mast cells (CD117⁺), mononuclear cells (CD11c⁺), plasma cells, activated-T cells (MUM1⁺), B cells, myeloid cells (PD1⁺) and neutrophilic granulocytes (myeloperoxidase⁺) compared with healthy donor specimens. We observed all of these immune cell markers in different types of lung cancers including squamous cell carcinoma, adenocarcinoma, adenosquamous cell carcinoma, small cell carcinoma, papillary adenocarcinoma, metastatic adenocarcinoma, and bronchioloalveolar carcinoma. The numbers of all tumor-associated immune cells (except MUM1⁺ cells) in stage III cancer specimens was significantly greater than those in stage I samples. We observed substantial stage-dependent immune cell infiltration in human lung tumors suggesting that the tumor microenvironment plays a critical role during lung carcinogenesis. Strategies for therapeutic interference with lung cancer microenvironment should consider the complexity of its immune cell composition.     

肿瘤抑素抗肿瘤相关肽的克隆及生物活性

为得到肿瘤抑素中具有直接抗肿瘤活性肽并检测其生物学活性,人工合成肿瘤抑素中185～2 0 3位氨基酸(19肽)所对应的核苷酸序列,将其连接到融合蛋白表达载体pTYB2中,酶切和测序鉴定后,转化到大肠杆菌BL 2 1(DE3)中诱导表达.表达的融合蛋白经几丁质亲和层析、二硫苏糖醇(DTT)的柱内还原,直接获得可溶性19肽.利用MTT法,细胞生长曲线,小鼠H2 2腹水型转移型肝癌实体瘤模型抑瘤实验并结合组织病理学切片,研究19肽的生物学活性.获得的19肽对B16小鼠黑色素瘤细胞、人SMMC 772 1肝癌细胞、人脐静脉内皮细胞的生长均具有抑制作用.小鼠H2 2腹水型肝癌抑瘤率达4 8 4 6 % .病理学切片显示,19肽可促使小鼠肿瘤组织坏死,血管数量减少.19肽具有较强的直接抗肿瘤活性,有可能成为肿瘤治疗的一种新的有前景的药物.     

Some differentiating effects of selenium on the cultured human hepatoma cells and human pulmonary adenocarcinoma cells in vitro

Two human hepatoma cell lines, QGY-7703 and SMMC-7721, and two human pulmonary adenocarcinoma cell lines, LTEP-a-2 and SPC-A-1, were found to respond to 1 μg/mL Na₂SeO₃, 24 h, in-vitro treatment by decreasing its confluent saturation density. The same treatment was found to cause an increase in the adhesiveness of cells measured as resistance to detachment by trypsin/EDTA. The pathological features of tumors after heterotransplantation of treated and untreated cells were similar, but the size of tumor grown from treated cells was much smaller.