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目的 观察骨骼肌细胞条件培养液 (MCM)体外对肿瘤细胞增殖的影响 ,并初步分析骨骼肌源性抑瘤物的理化特性及抑瘤机理。 方法 用噻唑蓝分析法 (MTT法 )分析新生大鼠MCM对不同肿瘤细胞系的体外抑瘤作用。用超滤分离、热灭活处理、胰酶消化、凋亡分析及形态学观察分析骨骼肌源性抑瘤物的理化特性及抑瘤机理。 结果 MCM与哺乳动物源性肿瘤细胞 (小鼠骨髓瘤SP2 / 0及Wistar大鼠癌肉瘤Walker2 5 6 )、人源性白血病细胞 (人慢性粒细胞白血病K5 6 2及人急性淋巴细胞白血病HL6 0 )、实体瘤细胞 (人结肠腺癌细胞LS 174 T及人前列腺癌细胞PC3 M) ,以及不同转移潜能肺癌细胞 (人肺癌低转移株PLA80 1 C及人肺癌高转移株PLA80 1 D)共同培养后 ,肿瘤细胞的增殖显著下降 (P <0 0 1～ 0 0 5 ) ,且肿瘤细胞增殖呈不同程度MCM浓度依赖性。MCM与正常细胞 (兔关节骺板细胞RGP 2 )共同培养后 ,细胞增殖无下降。同一来源肺癌细胞 ,在MCM稀释至原液的 6 2 5 %时 ,仍见高转移株增殖显著受抑 (P <0 0 1～ 0 0 5 ) ,而在MCM稀释至原液的 2 5 %时 ,低转移株增殖即无显著受抑。MCM的抑瘤活性存在于分子量 10 0 0 0以下超滤组分 ,热灭活后活性消失 ,胰蛋白酶消化后活性仍存在。MCM不引起K5 6 2细胞凋亡 ,而导致肿瘤细胞膜变?
Objective To observe the effects of conditioned medium (MCM) on the proliferation of tumor cells in vitro and to analyze the physico-chemical properties and anti-tumor mechanism of skeletal myogenic tumor suppressor. Methods MTT assay was used to analyze the antitumor effect of MCM on different tumor cell lines in vitro. The physicochemical properties and anti-tumor mechanism of skeletal myogenic tumor suppressor were analyzed by ultrafiltration, heat inactivation, trypsin digestion, apoptosis analysis and morphological observation. RESULTS: MCM was associated with mammalian-derived tumor cells (mouse myeloma SP2 / 0 and Wistar rat carcinosarcoma Walker 256), human leukemia cells (human chronic myeloid leukemia K562 and human acute lymphoblastic leukemia HL6 0 ), Solid tumor cells (human colon adenocarcinoma LS 174 T and human prostate carcinoma PC3 M), as well as co-cultures of different metastatic lung cancer cells (human lung cancer low metastasis PLA80 1 C and human lung cancer high metastasis PLA80 1 D) , The proliferation of tumor cells decreased significantly (P <0.01 ~ 0.05), and the proliferation of tumor cells showed different degrees of MCM concentration dependence. After MCM was cultured with normal cells (rabbit epiphyseal plate cells RGP 2), the cell proliferation did not decrease. In the same source of lung cancer cells, the proliferation of MCT was still significantly inhibited (P <0.01 ~ 0.05) when the MCM was diluted to 62.5% of the original solution, while when MCM was diluted to 25% of the original solution, The proliferation of low-transfer strains was not significantly inhibited. The antitumor activity of MCM exists in the ultrafiltration components with a molecular weight below 100 000, the activity disappeared after heat inactivation, and the activity still exists after trypsin digestion. MCM does not cause K562 cells apoptosis, leading to tumor cell membrane change?