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

An experimental research on inhibitory action of striated muscle condition medium on metastases of S-180 carcinoma cell

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。

Objective：To explore the effect of organic microenvironment of striated muscles on the S-180 carcinoma cell,and to investigate the mechanism of the rarity of cancer metastases in striated muscles.Methods：Primary culture of newborn Wistar rat cardiac and skeletal muscle cells were established,and the murine cardiac muscle conditioned medium（CMCM） and skeletal muscle conditioned medium（MMCM） was prepared to test its effect in vitro on S-180 by MTT assay.Cisplatin was used as positive control for CMCM and MMCM,DMEM was used as negative control.Murine benign renal cells-mesangial cells were used as negative control for S-180 carcinoma cells.The cardiac and skeletal muscle cells conditioned medium（1：1） was also tested boiling at 100℃ and treatment with trypsin.Results：The proliferation of tumor cell lines of S-180 carcinoma cell was significantly restrained when cultured with cardiac muscle cells conditioned medium and skeletal muscle cells conditioned medium（P 0.01）,while the murine benign renal cells were not affected（P 0.05）.Cisplatin restrained S-180 cells（P 0.05） and the murine benign renal cells（P 0.05）.The tumor suppressor was trypsin resistant but thermolabile.Conclusion：Straited muscle cells could selectively inhibite the proliferation of cancerous cells in vitro while benign cells are not affected.and the retraining mechanism of Cisplatin is different from straited muscle cells conditioned medium.The straited muscle derived tumor suppressor may play a key role in the rarity of tumor metastases in straited muscles.