【摘要】 目的 调节细胞培养液pH值,观察不同pH值对LAK细胞的增值和rIL-2、IFN-α2b、LAK细胞杀伤恶性细胞的影响。方法 采用淋巴细胞分离液梯度离心法从正常人外周血分离单个核细胞并用rIL-2激活,于96孔细胞培养板或培养管中培养。应用细胞计数方法观察pH值分别为6.8、7.3、7.6的培养液,对rIL-2激活单个核细胞的影响。用7404细胞作为靶细胞、MTT法测定培养液pH值分别为6.8、7.3、7.6状态下rIL-2、IFN-α2b的杀伤作用,以及IFN--α2b对ADM(阿霉素)的增效作用和LAK细胞的细胞毒性作用。结果 pH值为7.3、7.6状态下,有利于LAK细胞增殖,pH值为7.6状态下rIL-2、IFN-α2b、LAK细胞杀伤效果最佳;pH值为6.8状态下几乎无杀伤作用。pH值为6.8、7.6都不利于7404细胞生长,pH值为7.6有利于LAK细胞生长。pH值为6.8、7.3、7.6时,IFN-α2b都能增加ADM抗肿瘤作用,但pH值为7.6时,IFN-α2b+ADM杀伤效果最佳。结论 在偏碱性环境下,有利于LAK细胞增殖,并且在此状态下LAK细胞和rIL-2、IFN-α2b、IFN-α2b+ADM对肿瘤细胞杀伤效果最佳。更多还原

【Abstract】 Objective It has been verified that immunity factors, LAK cells, could kill malignant tumor cells(MTCS)in vitro.Why in most cases the growth of tumor cannot be effectively brought under control while the organ immunity system remains intact in function and reacts positively to tumor? Primary malignant tumor and metastatic tumor can be divided into prophase tumor of blood vessels and tumor of blood vessels. Prophase tumor of blood vessels is about (1-2) × 103μn in diameter, and it depends merely on diffusion to exchange substances with its host. Although tumor of blood vessels could exchange substances through capilliaries, serious low pH still exists in the environment of tumor cell groups which are more than 1.5 × 102 fun away from the capillaries. Further study demonstrates that extracellular pH 6.5 - 7.0 for the MTCS is the equivalent of 7.1 - 7.6 for the normal extracellular pH values. Earlier cell cultures were conducted in the solution of pH 7.3 - 7.4, and this could not reflect actual condition of solid tumor tissue in human being. It has been found for the first time that after perfusion with NAHCO3 solution through tumor a tery,rIL-2,IFN- α, ADM and so on were, perfused into primary carcinoma of liver and carcinoma of pancreas tissues, and surprising results were abtained. This experiment, by regulating the pH values of the cell culture intends to observe the differences of LAK cells proliferation and those of LAK cells and rIL-2 in killing the MTCs. Methods The single nucleus cells are separated from the normal human peripheral blood by way of gradient centrifugation of the lymphocyte separating agent, activated by rIL-2 and cultured in the cell culture dish with 96 holes or the culture tubes to observe how pH 6.8, 7.3 and 7.6 culture solutions affect the way that rIL-2 activates the single nucleus cells. With cells 7404 as the target cells, the killer role of rIL-2, INF-α2b, IFN-α2b + ADM and cytotoxin of LAK cells in the case of pH 6.8, 7.3 and 7.6 will be assayed one by one by the MTT method. Results Culture solution 7.3 or 7.6 favors the proliferation of LAK cells, and pH 7.6 yields the optimal lethality on the part of the LAK cells rIL-2, INF-α2b, INF-α2b + ADM while pH 6.8 yields almost no lethality . Neither pH 6.8 nor 7.6 culture solution favors the growth of cells 7404 but pH 7.6 is favorable to the growth of LAK cells. Conclusions pH values of a culture solution affect the proliferation of LAK cells and concerted role of LAK cells and IFN-α2b,rIL-2 against the malignant tumor. The low-alkali environment favors the LAK cells proliferation and provides the LAK cells,IFN-α2b,rIL-2,IFN-α2b+ ADM with optimal state for killing the tumor cells.更多还原