INDUCTION OF APOPTOSIS IN S-180 AND S-180R TUMOR CELLS BY ADRIAMYCIN IN VIVO

Apoptosis of tumor cells have become a new standard for chemotherapy. It is useful to demonstrate induction of apoptosis in tumor cells by anti-cancer drugs in vivo. We reported the results of apoptosis induction in murine tumor cell line S-180 and it's resistant cell line S-180R by adriamycin in different dose and different time. We found that apoptosis in S-180 cells could be induced by low dose of adriamycin, the apoptosis was started at 24 h. after the administration, and reached to 62.5% of the cells to apptosis until 72 h. Comparison with the parental cell line, only 13% of S-180R cells were apoptosed. At high dose, 20% of S-180R cells were apoptosed, whereas, almost all S-180 cells were killed in the same time. The lymphocytes were appeared in abdominal cavity of the mice after treatment of adriamycin for 24 h. It was very interested to find out that there was no lymphocyte left in the abdominal cavity of the mice with S-180R cells treated at high dose of adriamycin.

Overexpression of 14-3-3 protein protects pheochromocytoma cells against 1-methyl-4-phenylpyridinium toxicity

Objective To investigate the effects of 14-3-3 protein overexpression on the 1-methyl-4-phenylpyridinium （MPP^＋） induced pheochromocytoma （PC12） cell death and the potential mechanisms. Methods pcDNA3.1（＋）-14-3-3 plasmids, which could be expressed in mammalian cell, were constructed and transfected into PC 12 cells with Lipofectamine 2000. The expression of 14-3-3 protein, Bcl-2 protein, and BAD protein were determined by western blot. 3-（4,5- dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay, microplate reader, and flow cytometric analysis were used to measure cell viability, the caspase activity, and apoptotic ratio respectively. Results （1） The expression of 14-3-3 protein increased significantly three weeks after pcDNA3.1（＋）-14-3-3 plasmids transfected into PC 12 cells. （2） MPP^＋ caused a decrease of cell viability in a dose-dependent manner. At 100μmol/L MPP^＋, cell viability reduced approximately 50%. （3） The caspase activity increased along with the MPP^＋ concentrations rising and reached its maximum value （0.34 μmol/mg protein） at 100 μmol/L MPP＊. However caspase activity decreased significantly when the MPP^＋ concentration exceeded 100 μmol/L. （4） Overexpression of 14-3-3 protein decreased the apoptosis ratio of PC 12 cells treated with 100μmol/L MPP^＋ from 26.5% to 8.6%. （5） Bcl-2 protein tended to decrease but BAD protein tended to increase after treatment of PC 12 cells with 100 μmol/L MPP^＋. Overexpression of 14-3-3 protein significantly increased the cellular level of Bcl-2 protein and decreased that of BAD protein. Conclusion Overexpression of 14-3-3 protein may reduce MPP^＋-induced apoptotic cell death in PC12 cells by up-regulating the Bcl-2 expression and down-regulating the BAD expression. These results may provide a promising target for treatment of Parkinson＇s disease.

Preliminary Validation of Tumor Cell Attachment Inhibition Assay for Developmental Toxicants With Mouse S180 Cells

This study was designed to explore the possibility of using ascitic mouse sarcoma cell line (S180) to validate the mouse tumor cell attachment assay for developmental toxicants, and to test the inhibitory effects of various developmental toxicants. The results showed that 2 of 3 developmental toxicants under consideration, sodium pentobarbital and ethanol, significantly inhibited S180cells attachment to Concanavalin A-coaed surfaces. Inhibition was dependent on concentration, and the IC50 (the concentration tha reduced attachment by 50% ), of these 2 chemicals was 1.2×10-3mol/L and 1 .0 mol/L, respectively. Anoher developmental toxiant, hydmiortisone, did not show inhibitory activity. Two non-developmental toxicants, sodium chloride and glycine were also tested and these did not decrease attachment rates. The main results reported here were generally sindlar to those obtained with ascitic mouse ovdrian tumor cells as a model. Therefore, this study added further evidence to the conclusion that cell specificity does not lindt attachment inhibition to Con A-coated surfaces, so S180 cell may serve as an altemative cell model, especially when other cell lines are unavailable. Furthermore, after optimal validation, it can be suggested that an S180 cell attachment assay may be a candidate for a series of assays to detect developmental toxicants.

Inhibitory activity of polysaccharide extracts from three kinds of edible fungi on proliferation of human hepatoma SMMC-7721 cell and mouse implanted S180 tumor

AIM To determine the activities ofpolysaccharide extracts from Flammulina velutipes (Curt. ex Fr. ) Sing (FV), Lentinusedodes (LE) and Agaricus bisporus Sing (AB)on the proliferation of human hepatoma SMMC-7721 cells in vitro and on mouse implanted S-180tumors in vivo.METHODS The polysaccharide extracts were isolated from the fruit bodies of FV, LE and AB by the methods of hot-water extraction, Sevag’sremoval of proteins, ethanol precipitation,trypsin digestion and ethanol fractionalprecipitation. Human hepatoma SMMC-7721 cells were treated with 50 mg/L Polysaccharide extracts, and the mitosis index, mitochondria activity and cell proliferation were detected at different times in both control and experimental groups. The mice with S-180 implanted tumors were injected with the polysaccharide extracts at 24 mg/ kg body weight for 9 d and the tumorweight was measured on the 15th day.RESULTS The mitosis index of hepatoma cells in vitro could be significantly decreased by treatment with the polysaccharide extracts fromthe three kinds of edible fungi (P < 0 .005 ). Thecell numbers and mitochondria activity of SMMC7721 cells treated with polysaccharide extracts were lower than those in control groups (P <0.005). The inhibition rates of polysaccharide extracts against implanted S-180 tumors in mice were 52.8%, 56.6% and 51 .9% respectivelycompared with that in c0ntrol gr0ups.CONCLUSI0N The POIysaccharide extractsfrom the three kinds of edible fungi could inhibitnot only the Cultured malignant cells in vitfO butalso impIanted Sl80 tum0r i0 vivo.

Effects of natural cerebrolysin on protective proteins and pro-apoptotic molecules in mesenchymal stem cells following beta-amyloid peptide1-40-induced endoplasmic reticulum stress

BACKGROUND： Studies have demonstrated that β-amyloid peptide （Aβ）, a characteristic pathological product of Alzheimer＇s disease （AD）, results in neuronal endoplasmic reticulum stress （ERS）. However, the mechanisms of traditional Chinese medicine against ERS in AD are poorly understood. OBJECTIVE： To measure expression levels of protective proteins （GRP78 and GRP94） of ER molecular partners and pro-apoptotic Caspase-12 ER membrane expression following application of traditional Chinese medicine natural cerebrolysin （NC） to treat Aβ1-40-induced ERS. DESIGN, TIME AND SETTING： A parallel-controlled study was performed at the Institute of Integrated Western and Traditional Chinese Medicine, Shenzhen Hospital of Southern Medical University between September 2006 and November 2008. MATERIALS： Sprague Dawley male rats, 6-8 weeks old, were used to harvest tibial and femoral bone marrow. Isolation and purification of mesenchymal stem cells （MSCs） were established from the whole bone marrow by removing non-adherent cells in primary and passage cultures. Aβ1-40 was provided by Sigma, USA. NC was provided by Shenzhen Institute of Integrated Chinese and Western Medicine, China. NC was predominantly composed of Renshen （Radix Ginseng）, Tianma （Rhizoma Gastrodiae）, and Yinxingye （Ginkgo Leaf） in a proportion of 1 ： 2： 2. Following conventional water extraction technology, an extract （1 ： 20） was prepared. Six adult, male, New Zealand rabbits underwent intragastric administration of NC extract （0.976 g/kg per day） for 1 month to prepare NC-positive serum, and the remaining 6 rabbits received intragastric administration of physiological saline to prepare normal blank serum. METHODS： A total of 500 nmol/L Aβ1-40 was used to establish ERS models of primary cultured MSCs. AD cell models were incubated with different doses of NC-positive serum （2.5%, 5%, and 10%）. MSCs treated with normal blank serum served as normal blank controls. MAIN OUTCOME MEASURES： Reverse transcription-polymerase chain reaction and fluorescent immunocytochemistry were respectively used to measure mRNA and protein expression levels of GRP78, GRP94, and Caspase-12 in MSCs. RESULTS： Following Aβ1-40 exposure, mRNA and protein expression levels of GRP78 and GRP94, as well as Caspase-12, significantly increased （P 〈 0.05）, suggesting successful establishment of ERS models. Following NC-positive serum application, mRNA and protein expression levels of GRP78 and GRP94 in MSCs significantly increased （P 〈 0.05 or P 〈 0.01）. However, mRNA and protein expression levels of Caspase-12 significantly decreased （P 〈 0.05, or P 〈 0.01） compared with the ERS model group. These effects were dose-dependent. CONCLUSION： NC downregulated Caspase-12 expression and upregulated GRP78 and GRP94 expression in MSCs in a dose-dependent manner under the state of Aβ1-40-induced ERS.

DEVELOPMENT OF AN ADRIAMYCIN RESISTANT MURINE TUMOR S-180 CELL LINE IN VIVO

Adriamycin resistant cells were obtained from low dotage treated BABL/c mice Inoculated with S-180 cells. Resistance of these cells for adriamycin was 66-fold more than their parental cells. The resistance for a typical DNA topoisomerase Ⅱ inhibitor VP16 (Etopcaide) was increased 9 times. Overexpression of multidrug resistant gene (MDR gene) products, P-glycoproteins (P-1 70), was also demonstrated by immunohistochemistry. Furthermore, the ability of the resistant cells to reduce net cellular drug accumulation measured by flow fluorescence cytometry was 89-fold higher than their parental cells. These results support the hypothesis that the resistance of S-180R cells to adriamycin was mainly due to the overexpression of P-glycoproteins. The S-180R cells will be useful to select drugs or some other therapeutic strategies to overcome multidrug resistance in vivo.

Saponin from Tupistra chinensis Bak Inhibits NF-κB Signaling in Sarcoma S-180 Cell Mouse Xenografts

This study examined the effect of saponins from Tupistra chinensis Bak （STCB） on the growth of sarcoma S-180 cells in vitro and in mouse xenografts as well as the underlying mechanisms. Cell proliferation was assessed by MTT assay. Cell cycle distribution was determined by flow cytometry. Sarcoma S-180 tumor-bearing mice were treated with different doses of STCB with 10 μg/mL 5-fluorouracil （5-Fu） as a positive control. The activity of nuclear factor （NF）-κB was detected by gel mobility shift assay. The mRNA level of NF-κB was determined by real-time quantitative RT-PCR. The results showed that in vitro STCB inhibited the growth of S-180 cells in a concentration-dependent manner, which was accompanied by cell cycle arrest at S-phase. In vivo STCB significantly inhibited the growth of S-180 tumor mouse xenografts in a dose-dependent manner with apparent induction of cell apoptosis. Moreover, STCB inhibited the activity of NF-rd3 p65 and reduced the expression of NF-κB p65 mRNA in mouse xenografts. It was concluded that STCB inhibits the proliferation and cell cycle progression of S- 180 cells by suppressing NF-κB signaling in mouse xenografts. Our findings suggest STCB is a promising agent for the treatment of sarcoma.

抑瘤饮增加小鼠S180移植瘤巨噬细胞浸润及TNF-α和iNOS表达

目的动态研究中药复方抑瘤饮对小鼠S180移植瘤内巨噬细胞(Mφ)浸润及TNF-α和iNOS表达的影响,揭示其体内抗瘤免疫机制。方法Balb/c小鼠右腋皮下接种S180肿瘤细胞,24h后,抑瘤饮组小鼠每日灌服抑瘤饮,对照组小鼠每日灌服等量凉开水。分别于第10、20、30天和第40天杀鼠取瘤制成切片,免疫组化染色法检测肿瘤组织内Mφ浸润及TNF-α和iNOS表达,以图像分析系统对染色结果进行测定。结果所有4个时间点抑瘤饮组Mφ浸润均显著高于对照组;第10天时抑瘤饮组TNF-α和iNOS表达与对照组无差异,第20、30天和第40天时高于对照组。结论抑瘤饮体内抗瘤作用可能与增加肿瘤组织中Mφ浸润及TNF-α和iNOS表达有关。

盐酸洛拉曲克在体内、外对S-180细胞株的抗增殖作用

研究全合成新型胸苷合成酶抑制剂盐酸洛拉曲克在体内、外对S 1 80细胞株及正常人胚肾HEK2 93细胞的抗增殖作用 ;使用MTT法测定抑制率 ,以提高荷腹水瘤小鼠存活时间及荷实体瘤瘤重减轻情况为指标考察盐酸洛拉曲克对S 1 80所致肿瘤的治疗作用 .结果表明 :在体外盐酸洛拉曲克对S 1 80肿瘤细胞株有较强的细胞毒作用 ,对正常细胞HEK2 93抑制作用较弱 (P <0 .0 5 ) ;体内实验显示盐酸洛拉曲克可明显提高荷腹水瘤小鼠的存活时间 ,减轻荷实体瘤小鼠的瘤重 ,疗效与氟尿嘧啶 ( 1 0mg kg)相当 (P >0 .0 5 )或更好 (P <0 .0 5 ) .可见 ,盐酸洛拉曲克在体内、外对S 1 80肿瘤细胞有显著的抗增殖作用 。

Effects of Low Dose Radiation on Tumor Apoptosis, Cell Cycle and Apoptosis-Related Protein Bcl-2 in Tumor-Bearing Mice

To study the effects of low dose radiation (LDR) on tumor apoptosis, cellcycle progression and changes of apoptosis-related protein Bcl-2 in tumor-bearing mice. Methods:Male mice of Kunming strain were implanted subcutaneously with S180 sarcoma cells in the left inguenas an in situ experimental animal model. Seven days later, the mice were subjected to 75 mGywhole-body γ-irradiation. At 24 and 48 h after the irradiation, all mice were sacrificed. The tumorsizes were measured, and tumor cell apoptosis and cell cycle progression were analyzed by flowcytometry. The expression of apoptosis-related protein Bcl-2 and the apoptotic rate of tumor cellswere observed by immunohistochemistry and electron microscopy. Results: Tumors grew significantlyslower after LDR (P 【 0.05). The tumor cells were arrested in G1 phrase and the expression of Bcl-2protein decreased at 24 h. Apoptotic rate of tumor cells was increased significantly at 48 h afterLDR (P 【 0.01). Conclusion: LDR could cause a G1-phase arrest and increase the apoptosis of tumorcells through the low level of apoptosis-related protein bcl-2 in the tumor-bearing mice. Theorganized immune function and anti-tumor ability are markedly increased after LDR. Our studyprovides practical evidence of clinical application to cancer treatment.

蟾蜍毒素对S180荷瘤小鼠肉瘤细胞Mg^(2+)-ATP酶、葡萄糖-6-磷酸酶活性的影响

目的探讨醇溶性蟾蜍毒素对S180荷瘤小鼠肉瘤细胞Mg2+-ATP酶(Mg2+-ATPase)、葡萄糖-6-磷酸酶(G-6-Pase)活性的影响。方法乙醇溶解蟾蜍分泌物原浆,采用减压蒸馏等方法,进一步部分分离纯化,获得醇溶性的蟾蜍毒素混合物(Ethanol extract of toad venom,简称EET),将其作用于S180荷瘤小鼠模型,计算肿瘤抑制率,应用酶化学染色法观察S180肉瘤细胞酶活性的变化。结果蟾蜍毒素组和氟脲嘧啶组的肿瘤抑制率分别为35.6%和36.9%(P<0.05),细胞质膜标志酶Mg2+-ATPase、内质网标志酶G-6-Pase反应颗粒变小,数量减少,密度变低,提示酶活性明显下降。结论蟾蜍毒素可降低S180荷瘤小鼠肉瘤细胞质膜标志酶Mg2+-AT-Pase及内质网标志酶G-6-Pase的活性,抑制肿瘤细胞的生长。

壮药白莲Ⅰ号方对荷瘤S_(180)小鼠抑瘤率及血浆白细胞介素-18的影响

目的观察壮药白莲Ⅰ号方对小鼠移植性肉瘤S180的抑瘤作用和其对小鼠血浆白细胞介素-18(IL-18)的影响,以探讨该药抑制肿瘤的作用是否与诱生小鼠血浆中IL-18的含量有关。方法建立小鼠移植肉瘤S180模型。用大、中、小剂量壮药白莲Ⅰ号方及环磷酰胺治疗肉瘤S180小鼠,观察其抑瘤率及血浆IL-18的含量。结果白莲Ⅰ号方在一定剂量范围内对荷瘤小鼠的抑瘤率>30%,并且白莲Ⅰ号方各剂量组均有促进荷瘤小鼠IL-18生成的作用趋势,改善荷瘤小鼠的生存质量。结论白莲Ⅰ号方能有效抑制小鼠S180实体瘤的生长,存在量-效关系;白莲Ⅰ号方具有诱生荷瘤S180小鼠血浆IL-18的作用,具有良好的免疫调节作用。

柴胡提取物对人肝癌细胞和小鼠S-180肉瘤的抑制作用

目的 :研究柴胡水提物对体外培养的人肝癌 SMMC-772 1细胞线粒体代谢活性、细胞增殖及小鼠移植性 S-1 80实体瘤的影响。方法 :中药柴胡 ( BCDC)经粉碎、浸提、超速离心、减压浓缩、冷冻干燥得柴胡提取物。人肝癌 SMMC-772 1细胞经不同浓度柴胡提取物处理不同时间后 ,检测了各实验组和对照组细胞的有丝分裂指数、细胞增殖及线粒体活性几方面的变化。S-1 80肉瘤小鼠连续 9d腹腔注射 3 3 .3 mg/ kg体重 BCDC提取物 ,停药 1 5d后 ,杀死小鼠称量瘤重。结果 :1经 3 55mg/ L BCDC提取物处理人肝癌 SMMC-772 1细胞 2 4 h,其线粒体活性为对照组的 50 .2 6% ,处理 72 h为 2 4 .82 %。2以 3 55mg/ L BCDC提取物处理 7d中 ,人肝癌细胞数分别是血清对照组( FCS)的为 57.5% ,4 5.88% ,3 7.0 6% ,2 8.3 7% ,2 0 .90 % ,1 2 .89% ,1 4.1 8% ;无血清对照组 ( NON)的 65.0 9% ,62 .2 4 % ,56.99% ,4 9.0 8% ,4 0 .78% ,2 6.78% ,2 8.4 1 %。 3经 BCDC提取物处理 2 4 h后 ,即使更换正常培养基 ,人肝癌细胞数在 96h的恢复培养中仍低于处理前的细胞数。 4有丝分裂指数实验组为 0 .3 0 ,对照组分别为 5.3和5.4 ( P<0 .0 0 1 )。 5BCDC提取物对小鼠 S-1 80实体瘤抑制率为 87.2 1 %。

电脉冲致离体S-180细胞电穿孔的观察与研究

扫描电镜下我们观察到了一定强度电脉冲致 S- 180细胞电穿孔的现象。同时分别改变电压、电容、脉冲个数 ,对 S- 180离体细胞进行电脉冲作用 ,通过苔盼蓝追踪 ,发现在一定的条件下 ,随着对 S- 180离体细胞电穿孔的电压越大、电容越小、脉冲个数越多 ,其穿孔百分率越大。

β-葡聚糖对S180荷瘤小鼠实体瘤抑制作用及其机制研究

目的研究β-葡聚糖对S180荷瘤小鼠肿瘤抑制作用及其机制。方法将S180细胞接种于小鼠右上肢皮下建立S180荷瘤小鼠模型,随机分为模型组、顺铂组(0.2 mg/kg,腹腔注射)、β-葡聚糖低、中、高剂量(50、100、200 mg/kg)组,每组16只,雌雄对半。隔日给药,连续21 d,ELISA法检测瘤组织、血液及肝脏相关指标表达,Western Blotting检测凋亡蛋白Caspase-3的表达。结果β-葡聚糖三个剂量组均具有明显的抑瘤作用(P<0.01)。与模型组相比,β-葡聚糖三个剂量组脾脏指数均升高(P<0.01);β-葡聚糖三个剂量组AST、ALT、UA和CRE含量下降(P<0.01)。与模型组相比,β-葡聚糖三个剂量组均能增加凋亡蛋白Caspase-3蛋白的表达(P<0.05)。结论β-葡聚糖显著抑制S180腹水瘤荷瘤小鼠的肿瘤生长,其作用机制可能与促进肿瘤细胞的凋亡相关。

灵芪胶囊对S_(180)荷瘤小鼠IL-1和IL-6蛋白表达的影响

目的:研究灵芪胶囊对S_(180)荷瘤小鼠的抗肿瘤作用,并探讨其可能的作用机制。方法:将S_(180)瘤株接种于小鼠右前肢腋部皮下建立动物模型,并采用ELISA法检测小鼠外周血IL-1和IL-6蛋白含量。结果:灵芪胶囊能明显增加S_(180)荷瘤小鼠外周血IL-1和IL-6蛋白含量,能改善和增强荷瘤小鼠的免疫功能。结论:灵芪胶囊对S_(180)荷瘤小鼠有明显抑瘤作用,其作用机制可能是通过改善小鼠免疫功能而实现的。

山仙颗粒对S-180荷瘤小鼠T细胞活性及Caspase-3的影响

目的:探讨山仙颗粒(SXG)对S-180荷瘤小鼠T细胞活性及Caspase-3的影响。方法:将60只昆明种小鼠随机选取12只作为空白对照组,其余48只建立S-180荷瘤小鼠模型后随机分为模型组及SXG低(0.025 g/mL)、中(0.05 g/mL)、高(0.1 g/mL)剂量组各12只,空白对照组、模型组每日早晚灌胃生理盐水0.4 mL,其余各组每日早晚灌胃相应剂量药物,连续30天。末次给药24小时后,采用颈椎脱臼法处死小鼠,游离脾脏,剥离肿瘤组织;应用MTT比色法及免疫组织化学法检测荷瘤小鼠脾脏T细胞毒效应及S-180实体瘤组织中Caspase-3的表达。结果:SXG低、中、高剂量组小鼠脾脏T细胞对于S-180荷瘤小鼠细胞的抑制率高于模型组(P<0.01),且与SXG浓度呈正相关;各组效应细胞与靶细胞的比例不同,对S-180荷瘤小鼠细胞的抑制作用也不同(P<0.01),且与T细胞浓度呈正相关。SXG低、中、高剂量组S-180荷瘤小鼠肿瘤组织中Caspase-3表达率高于模型组(P<0.01),且与SXG浓度呈正相关。结论:SXG具有抑制肿瘤生长的作用,其机制可能与其增强T细胞活性进而抑制肿瘤细胞增殖及促进Capase-3介导的肿瘤细胞凋亡有关。

人胎肝中低分子天然抑瘤物对小鼠肉瘤S-180作用的研究

从人胎儿肝脏中分离出一种分子量较小的抑瘤物质(FLS-MeOH),观察了它对小鼠肉瘤S-180细胞生长的抑制效应。在体外琼脂培养中,当FLS-MeOH浓度达到300μg/ml时,可完全抑制S-180细胞的集落形成;给荷瘤小鼠每日注射FLS-MeOH8mg/g体重,共20d,可明显延长小鼠的存活时间,部分小鼠可以无病存活。这些结果说明FLSMeOH在体内外均有明显的抗肿瘤活性,而且具有分子量小、无组织和种系特异性,以及对肿瘤细胞不可逆的毒性作用等特性。

珍珠梅提取物对S_(180)小鼠血管内皮生长因子及血管生成素蛋白-1表达的影响

目的研究珍珠梅水提物对S180荷瘤小鼠体内血管内皮生长因子(VEGF)、血管生成素蛋白-1(Ang-1)蛋白表达的影响。方法取60只小鼠复制S180荷瘤模型,随机分成5组:珍珠梅水提取物大、中、小剂量组,对照组,环磷酰胺组。观察珍珠梅水提取物对S180荷瘤小鼠的抑制率;应用免疫杂交方法检测肿瘤组织中VEGF、Ang-1蛋白的表达。结果珍珠梅水提取物对S180荷瘤小鼠有明显的抑制作用(P<0.05),免疫杂交结果显示,珍珠梅水提取物可以明显降低VEGF蛋白在肿瘤组织中的表达(P<0.05);Ang-1的表达在各组中无显著性差异。结论珍珠梅水提物可以抑制S180荷瘤小鼠的肿瘤生长,下调VEGF蛋白在肿瘤组织中的表达,抑制肿瘤血管生成。

横纹肌细胞培养液对S-180细胞生长抑制作用的体外实验研究

目的:观察横纹肌细胞微环境对肿瘤细胞生长的影响,探讨横纹肌内转移瘤罕见这一现象的发生机制。方法:采用酶解法原代培养新生Wistar大鼠心肌细胞和骨骼肌细胞,形态学和免疫酶染色方法检测横纹肌α-肌动蛋白进行细胞鉴定,免疫酶染色法进行细胞纯度鉴定,MTT法分析比较骨骼肌和心肌细胞细胞培养液对S-180细胞和正常肾小球系膜细胞的体外生长抑制作用,并运用胰酶消化法,热灭活法初步探讨横纹肌细胞培养液中抑瘤物质的理化性质。结果:S-180细胞与CMCM、MMCM共同培养后,细胞存活率明显下降,与DMEM相比差异均具有显著性(P<0.05),MCs细胞的细胞存活率无明显影响(P>0.05)。DDP对S-180细胞和MCs细胞均具有显著性抑制作用(P<0.05)。心肌细胞培养液和骨骼肌细胞培养液经胰酶消化后抑制活性仍然存在(P<0.05),经热灭活后活性消失(P>0.05)。结论:新生大鼠横纹肌细胞培养液对S-180细胞的增殖有明显的抑制作用;而对正常细胞的增殖无明显影响,与顺铂的抑制作用明显不同。横纹肌细胞培养液中有可能存在一种抑瘤物质,可能是横纹肌转移瘤罕见的关键因素。