CLONING AND SEQUENCING OF IMMUNOGLOBULINVARIABLE－REGION GENE OF A MONOCLONALANTIBODY SPECIFIC FOR HUMANHEPATOCARCINOMA

ＣＬＯＮＩＮＧＡＮＤＳＥＱＵＥＮＣＩＮＧＯＦＩＭＭＵＮＯＧＬＯＢＵＬＩＮＶＡＲＩＡＢＬＥ－ＲＥＧＩＯＮＧＥＮＥＯＦＡＭＯＮＯＣＬＯＮＡＬＡＮＴＩＢＯＤＹＳＰＥＣＩＦＩＣＦＯＲＨＵＭＡＮＨＥＰＡＴＯＣＡＲＣＩＮＯＭＡＹａｎｇＰｉｎｇ杨萍；ＧａｏＬｅｉ高...     

QUALITATIVE STUDY OF SIALOMUCINS CHANGES DURING N－METHYL－N－NITROSOUREA－INDUCED C OLONIC CARCINOGENESIS IN MICE

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ISOLATION AND CHARACTERIZATION OF AN ADRIAMYCIN－RESISTANT SUBLINE OF THE HUMAN GASTRIC ADENOCARCINOMA CELLLINE

ＩＳＯＬＡＴＩＯＮＡＮＤＣＨＡＲＡＣＴＥＲＩＺＡＴＩＯＮＯＦＡＮＡＤＲＩＡＭＹＣＩＮ－ＲＥＳＩＳＴＡＮＴＳＵＢＬＩＮＥＯＦＴＨＥＨＵＭＡＮＧＡＳＴＲＩＣＡＤＥＮＯＣＡＲＣＩＮＯＭＡＣＥＬＬＬＩＮＥＷａｎｇＹａｎｐｉｎｇ；王艳萍；ＸｕＧａｎｇ；徐刚（Ｉ...     

EXPRESSION OF c－myc GENE AND BIOSYNTHESIS OF BIOLOGICAL MACROMOLECULES IN ANTISENSE TRANSFECTANT HL_（60）~R－9

ＥＸＰＲＥＳＳＩＯＮＯＦｃ－ｍｙｃＧＥＮＥＡＮＤＢＩＯＳＹＮＴＨＥＳＩＳＯＦＢＩＯＬＯＧＩＣＡＬＭＡＣＲＯＭＯＬＥＣＵＬＥＳＩＮＡＮＴＩＳＥＮＳＥＴＲＡＮＳＦＥＣＴＡＮＴＨＬ_（６０）~Ｒ－９ＬｉＹｉｎｘｉｏｎｇ李尹雄，ＦａｎＭｕｚｈｅｎ范慕贞，Ｚｈａ...     

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ＴＵＭＯＲＮＥＣＲＯＳＩＳＦＡＣＴＯＲ－αＡＬＴＥＲＳＰＲＯＴＥＩＮＭＥＴＡＢＯＬＩＳＭＡＮＤＣＥＬＬ－ＣＹＣＬＥＫＩＮＥＴＩＣＳＩＮＭＡＬＩＧＮＡＮＴＴＵＭＯＲＹｅＳｈｅｎｇｌｏｎｇ叶胜龙；ＴａｎｇＺｈａｏｙｏｕ汤钊猷；ＢｒｕｃｅＲＢｉｓｔｒｉａｎ...     

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ＴＨＥＥＸＰＲＥＳＳＩＯＮＯＦＣ－ＭＹＣＡＮＤＮ－ＲＡＳＯＮＣＯＧＥＮＥＳＩＮＨＵＭＡＮＨＥＰＡＴＯＣＥＬＬＵＬＡＲＣＡＲＣＩＮＯＭＡ－ＡＮＩＮＳＩＴＵＨＹＢＲＩＤＩＺＡＴＩＯＮＳＴＵＤＹＯＮＰＡＲＡＦＦＩＮＥＭＢＥＤＤＥＤＴＩＳＳＵＥＳＥＣＴＩＯＮ...     

THE EXPRESSION OF CATHEPSIN－D，C－erbB－2 AND EGFR IN BREAST CANCER AND ITS CORRELATION TO LYMPHATIC ME

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Biologic effects of Tween 80 in combination with hyperthermia on human stomach cancer cell line BGC-823]

Cells of human stomach cancer cell line BGC-823 were treated with Tween 80 and hyperthermia 39 degrees C to 43 degrees C for 20 to 100 minutes. Normal human fibroblasts and mitomycin C were used as controls. The results showed Tween 80 greatly reduced the activation energy of BGC-823 cells. Synergistic effect was observed if applied with heat 39 degrees C. With the increase in temperature and time, the inhibitory effect on the cancer cells was gradually intensified. The lethal rate of heat 41 degrees C associated with Tween 80 was 5.2 times as that of heat alone. The inhibitory effect of heat 41 degrees C for 100 minutes combined with Tween 80 was equivalent to 43 degrees C for 100 minutes. In other words, the heat critical Temperature of BGC-823 cells was reduced about 2 degrees C. The measurement of membrane fluidity, SDH activity and the rest also showed heat 41 degrees C produced the highest synergy with Tween 80. The specific and sustained action exceeded that of mitomycin C. The response of the normal cells was mild and reversible. These studies suggest that the synergistic mechanism of Tween 80 and hyperthermia possibly lies on the cell plasma membrane system.     

The enhancement of tween-80 on the antitumor effect of the hyperthermia 41°C in tumor-bearing mice

B16 melanoma cells were inoculated into the BALB/C mice to establish melanoma-bearing models. The antitumor effect of Tween-80 in combination with hyperthermia 41°C was studied. We observed the changes of the mortality of tumor-bearing mice, the tumor growth curves, activities of serum tumor necrosis factor (STNF) and the level of serum sialic acid (SSA) in tumor-bearing mice. The number of pulmonary metastatic tumor foci from blood flow was also detected. The results showed that combined with Tween-80, hyperthermia 41 °C could dramatically suppress the growth of the melanoma in the feet of mice, survive the tumor-bearing mice and decrease the number of pulmonary metastatic tumor foci but no significant effects were observed by treatment with Tween-80 or hyperthermia at 41 °C alone. The activities of STNF and the level of SSA of the melanoma-bearing mice kept at higher levels than those of normal BALB/C mice. Tween-80 combined with heating 41°C significantly decreased activities of STNF, but the level of SSA still kept high level within 1 to 2 weeks and decreased 10 weeks later with the tumor regression. These results demonstrate that Tween-80 may make hyperthermia exert effective antitumor effect below the critical temperature and increase the safety of hyperthermia in treatment. It is one of the most ideal synergist with hyperthermia. The changes of STNF and SSA suggest that the synergetic effect could involve in facilitating the exposure of tumor antigen and activation of immune system. This work is supproted by National Nature Science Fund (No. 39170825) and Shanghai Science Developing Fund.     

吐温80对温热抑瘤作用的增强效应——小鼠黑色素瘤实验研究

 目的: 研究吐温80合并41℃温热的抗肿瘤效应。方法: 接种B16黑色素瘤细胞至BALB/C小鼠不同部位建立荷瘤鼠模型, 观察荷瘤鼠死亡率、肿瘤生长曲线、血清肿瘤坏死因子和唾液酸的变化以及经血行转移的肺部瘤灶数的变化。结果: 吐温80合并41℃温热能明显地抑制小鼠黑色素瘤的生长, 延长荷瘤小鼠平均存活时间, 使肺部的转移瘤灶数大大降低；但吐温80和温热单独作用时均无此效果。荷瘤小鼠血清肿瘤坏死因子(sTNF)活性和唾液酸水平均明显高于正常BALB/C小鼠, 吐温80合并41℃、100分钟温热可显著降低sTFN活性, 并引起血清唾液酸水平的进一步升高。处理后第10周, 随着肿瘤的缩小, 唾液酸水平有明显的下降。结论: 吐温80能使温热在低于临界温度时即可发挥有效的抗肿瘤效应, 明显提高温热治疗的疗效和安全性, 是一较理想的温热合并药物。血清TNF和唾液酸的变化提示协同效应可能与肿瘤抗原的暴露和机体免疫抗肿瘤作用的活化亦有关。     

吐温80合并41℃温热对荷瘤小鼠的抑瘤效应

用B16黑色素瘤接种于小鼠后脚掌部皮下或腹腔内．待成实体瘤后经尾静脉注入吐温80．并局部温热41℃。观察实体瘤的生长曲线及荷瘤动物的半数存活时间及半数致死时间．还观察吐温80．41℃及两者合并作用的B16细胞经血行播散后肺内成瘤灶数的变化。实验结果表明：单独吐温或41℃温热对实体瘤的生长，荷瘤小鼠的存活时间及血行播散肺内成瘤灶数均无明显影响．而两者合并作用可明显抑制实体瘤的生长，并明显延长荷瘤小鼠的生存时间及减少血行播散肺内成瘤灶数。实验证明；吐温80合并41℃温热有明显的实体瘤抑瘤效应．而且较安全，为开展新的温热肿瘤治疗提供动物实验依据。     

Enhancement of nitrocaphane cytotoxicity by hyperthermia in vitro

HEp-2 cells were treated with hyperthermia (39–44°C) and nitrocaphane (NC) at various time intervals. A 1 h exposure to 39°C and 41 °C was non-lethal to cells, but it did potentiate the cell killing of NC (1.0 μg/ml). It was further shown that the sequence between heat and the drug can affect cell survival. Cell killing effect was decreased when heat was given before or after administration of drug. In contrast the simultaneous administration of these two modalities was synergistic. Maximal thermotolerance of HEp-2 cells was developed using an 8-h interval at 37 °C between the cell exposures to two equal thermal doses (44°C, 30 min). HEp-2 cells became thermotolerant when preheated for 30 min at 44°C followed by a 10-h interval at 37°C. The thermotolerant cells showed resistance to subsequent heat at 44°C (D₀=2.26 h, control D₀=0.38 h), to subsequent NC treatments, and to heat combined with NC. However, in the thermotolerant cells, cytotoxicity of NC was still enhanced by hyperthermia.     

Synergism between alkylating agent and cis-platin with moderate local hyperthermia: the effect of multidrug chemotherapy in an animal system

The combined effect of multidrug chemotherapy given in combination with hyperthermia was investigated using early-generation isotransplants of a spontaneous fibrosarcoma, FSa-II in C3Hf/Sed mice. Combinations of various types of chemotherapeutic agents, including alkylating agents, cyclophosphamide (CY) and 1, 3-bis(2-chloroethyl)-1-nitro-sourea (BCNU); antibiotics, bleomycin (BLM) and mitomycin C (MMC); an antimetabolite, 5-fluorouracil (5FU); and a platinum complex, cis-diamminedichloro-platinum(II) (cDDP), were examined using the tumour growth (TG) time assay. Simultaneously, the effect of glucose on the response to thermochemotherapy was investigated. Graded doses of the multidrugs were given i.p. immediately before hyperthermia with or without a glucose dose of 5 g/kg given i.p. 60 min before hyperthermia. Hyperthermia was given by immersing the tumour-bearing murine feet into a water bath set at 41·5 ± 0·05°C for 60 min. Dose-response curves were obtained between the TG time and drug dose. The thermal enhancement ratio (TER) was expressed as a ratio of the slope of the dose-response curve obtained at 41·5°C to that obtained at room temperature. To evaluate normal tissue damage, the number of white blood cells (WBC) was counted from a day before treatment to the 21st day after treatment. A substantial thermal enhancement of the anti-tumour effect was observed in all five multidrug regimens tested. Glucose administered prior to thermochemotherapy further enhanced the antitumour effect. The TER was largest for the combination of CY+cDDP (TER was 5 without glucose). The second largest TER was obtained for a combination of CY+cDDP+MMC (TER was 4·1 without glucose and 6·5 with glucose). The antitumour effects of these two combinations were synergistic at a test elevated temperature only. No synergistic effect was found at room temperature for any of the drug combinations tested. The smaller TERs were observed in the treatment regimens that included SFU. In general, a decrease in the number of WBC following multidrug chemotherapy was slightly less than that following the individual drugs.     

Defect in the development of thermotolerance and enhanced heat shock protein synthesis in the mouse temperature-sensitive mutant ts85 cells upon moderate hyperthermia

The effect of exposure to moderate hyperthermia on the induction of thermotolerance and heat shock protein (HSP) synthesis was investigated using mouse FM3A cells and the temperature-sensitive mutant ts85 cells. The thermal sensitivity of the two cell lines was markedly different; the mutant ts85 cells were more sensitive than the parental wild-type FM3A cells to heating at 41 and 44°C. The shift-up treatment of FM3A cells for 3 h at 39 5°C from 33°C induced thermotolerance development to subsequent heating at 44°C, with little if any enhancement of major HSP synthesis. On the other hand, the similar treatment of ts85 cells at the non-permissive temperature of 39 5°C induced significantly enhanced HSP synthesis, but could not induce thermotolerance. The exposure to 41°C also induced thermotolerance in the wild-type cells, but failed to induce tolerance in the mutant ts85 cells. These results suggest that enhanced major-HSP synthesis is neither a sufficient or necessary condition for thermotolerance development upon moderate heat shock. The mechanism of thermotolerance is discussed by relating the observed defect in thermotolerance development to the known defect in ubiquitindependent protein degradation system of the mutant ts85 cells at non-permissive temperature.     

Hyperthermic modulation of SN‐38‐induced topoisomerase I DNA cross‐linking and SN‐38 cytotoxicity through altered topoisomerase I activity

The effect of different temperatures (37–42.5°C) on SN‐38 (the active metabolite of CPT‐11) cytotoxicity was examined in the human lung carcinoma cell lines H460 and Calu‐6 as well as the murine fibrosarcoma cell line L929. The cytotoxicity of SN‐38, determined by MTT cell survival assay, was significantly increased in each cell line in combination with 41.8°C hyperthermia (×60–120 min); the combination of SN‐38 with 40.5°C and 42.5°C, however, was unchanged compared to 37°C. Determination of topoisomerase (Topo) I DNA cross‐linking in Calu‐6 cells and L929 cells after treatment with SN‐38 showed the same temperature profile as seen in the cell‐survival assays with increased Topo I DNA cross‐linking after treatment with the combination of SN‐38 and 41.8°C hyperthermia and unchanged Topo I DNA cross‐linking at 40.5°C and 42.5°C. To test the hypothesis that increased Topo I DNA cross‐linking and SN‐38 cytotoxicity at 41.8°C is caused by hyperthermia‐modulated changes in Topo I activity, catalytic activity of Topo I extracted from each cell line and of purified human Topo I was determined at 20–42.5°C. Topo I activity was found to be gradually increased with rising temperatures, resulting in significantly higher activity at 41.8°C compared to 37°C; further increase of temperature past 41.8°C decreased Topo I activity back to levels found at 37°C. Our data are used to explain a series of events resulting in hyperthermic enhancement of Topo I DNA cross‐linking and SN‐38 cytotoxicity in combination with 41.8°C hyperthermia via increased Topo I activity. Int. J. Cancer 80:104–109, 1999. © 1999 Wiley‐Liss, Inc.     

Sensitizing effects of gallium citrate on hyperthermic cell killing in vitro

The lethal effects of gallium citrate in combination with heat were studied using four cell lines, L5178Y, FM3A, P388 and HeLa. Cells were incubated with different concentrations (0.2-2mM) of gallium citrate at 37°C for 24h and heated at a range of temperatures from 40-44°C for various time periods up to 6 h in the absence of gallium citrate. Survival and cell viability were determined by clonogenic assay and the dye-exclusion test, respectively. All of the cell lines tested were insensitive to heat below 41°C, but were very sensitive to heat above 43°C. Gallium citrate was cytotoxic to these cell lines at different levels: P388 and HeLa were far more sensitive than L5178Y and FM3A. The killing effects of heat at 41°C were greatly enhanced by gallium citrate in L5178Y and P388 cells. The Arrhenius analysis for the lethal effect of heat, determined by clonogenic assay, in L5178Y cells showed that the transition temperature was remarkably decreased for the gallium-treated cells from approximately 43°C to 41°C. The mechanism for this decrease in the transition temperature may be attributable to the additional effects of gallium citrate on energy metabolism. Preincubation with 0.05mM gallium citrate at 37°C for 7 days also enhanced heat sensitization at 41°C in L5178Y. This preincubation condition may correspond to the condition for the continuous infusion of gallium that is clinically used for cancer treatment. In contrast, treatment with gallium did not greatly enhance the sensitivity of FM3A or HeLa cells to heat at 41°C, but the effects of gallium were significant.     

Sensitization to cisplatin action by step-down heating in cddp-sensitive and -resistant cells

Hyperthermia treatment (≥ 43°C) has been shown to be able to (partially) reverse acquired cDDP resistance. However, such heat treatment is difficult to achieve in the clinic. Short pre-treatment at a high temperature (>42°C), immediately before a treatment at a lower temperature (<42°C) can enhance the heat toxicity of the lower temperatures. This “step-down heating schedule” was explored for its possible drug-sensitizing potential in in vitro-cultured cDDP-sensitive and -resistant murine and human tumour cells. A 10-min pre-treatment at 44°C enhanced the cytotoxicity of 41°C hyperthermia alone. It also enhanced sensitivity to cDDP when given at 37°C. However, it did not increase the 41°C-induced cDDP sensitization. Thus, no correlation was found between heat kill and cDDP sensitization for step-down heating schedules. The observed effects of step-down heating were comparable in sensitive and in resistant cells, so the step-down heating schedule, unlike the 43°C treatment, did not lead to a decrease of the cDDP-resistance factor. Yet the total cytotoxicity caused by this treatment protocol was 10-fold more than for cDDP with 41°C alone, due to the extra hyperthermic cell killing and the cDDP-sensitizing effect of the pre-treatment. This treatment could have a substantial impact on cDDP efficacy in the clinic even when cDDP resistance has developed. © 1995 Wiley-Liss, Inc.