MTT法检测裸鼠实体瘤药物敏感试验研究

Study of Chemosensitivity of Solid Tumor inNude Mice by MTT

本研究用一种快速、徽量ＭＴＴ比色法对分离得当的实体癌细胞进行药敏分析。检测中，ｐＨ１０．５缓冲液添入ＤＭＳＯ（１０％）溶解ＭＴＴ－甲 产物，使高达５ｘ１０４的ＢＥＬ－７４０２或Ｍ－ＧＣ８０－３癌细胞与甲 吸收值（２．３～２．５）有较好线性关系。辅以机械消化、用胰蛋白酶一胶原酶分离液分离异种移植瘤、经不同孔径滤器滤得单个瘤细胞悬液。癌细胞及其移植瘤分离的细胞在ＡＤＭ、Ｅ－ＡＤＭ、ＣＤＤＰ、５Ｆｕ、ＭＭＣ、ＭＴＸ、ＶＣＲ及ＰＹＭ（１ＰＰＣ／ｍｌ）各药连续作用下经ＭＴＴ和液闪法检测，两株癌细胞对１ＰＰＣ的ＡＤＭ、Ｅ－ＡＤＭ、ＣＤＤＰ、５Ｆｕ及ＭＭＣ等各显示半致死量药理作用，我们成功地用ＭＴＴ法对它们及其移植瘤分离的细胞进行药敏分析。８种药在１０ＰＰＣ／ｍｌ、１ＰＰＣ／ｍｌ时对它们各自细胞的药敏作用符合率高。ＢＥＬ－７４０２细胞及其移植瘤分离的细胞对５种药物（１ＰＰＣ各显示明显细胞毒作用；８种药物中的６种（１ＰＰＣ／ｍｌ）对ＭＣｃ８０－３细胞及其秽植瘤分离的细胞各有明显ＩＤ５０作用。比较了ＭＴＴ和液闪检测结果，两株癌细胞及其分离的移植瘤细胞对一组药物（８种、ＩＰＰＣ／ｍｌ）的药敏符合率达９１．５％。这提示了半自动化?

AbstractA rapid collormetric inicrotiter assay by MTT was used in this studyto analysis the chemosensitivity of sollid tumor cells disaggregatedproperly.In MTT assay, addition of a glycine buffer at PH 10.5 in DMSO( 10%) to solubitized MTT-formazan products resulted in a clear linearrelationship hetween MTT-formazan absorbed reading of 2.3￣2.5 and cellnumber up to 5x10 per well in BEL-7402 or MCC 80-3 cells.As a result,it overcomed most of the variability of absorbed reading caused by thepresence of a little culture medium with additional mechanic disaggrega-tion, single-tumor cell suspension was obtained by emzymatic digestion,including trysin-collagenase treatment , from the heterotransplantedtumors and filtering by the holders with different pore size.The tumorcell lines ( BEL-7402 and MGC 80-3 ) and their xenografts were used tocompare between the MTT and 3H-TdR scintillation assays followingcontinuous exposure to each agent for 6 days, ADM, E-ADM, CDDP, 5-Fu, MMC, MTX, VCR and PYM at 1 X PPC/ml concentration.Excellentcorrelation was observed for ID50 following the effect of ADM and theothers at 1 X PPC/ml on two cell lines analysed by two assays.Thechemosensitivity of two cell lines and their disaggregated xenograftswere analysed successfuly by MTT assay having a good correlation byexposing tlie cells to 8 drugs at 10 PPC/ml individually as well as 1 xFPC/ml, 0.1 PPC/ml, In these assays, two cell lines and the cellsdigested from xenografts were shown the similar cytoxicity effects ( ID50)