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目的：检测本实验室构建的表达人细胞色素 Ｐ４５０ ３Ａ４同功酶转基因细胞系 ＣＨＬ－３Ａ４的硝苯吡啶氧化酶活性。方法：利用多药耐药细胞Ｋ５６２ｒ的阿霉素（ＡＤＲ）耐药性可被硝苯吡啶逆转，而硝苯吡啶又可特异地被 ＣＹＰ３Ａ４代谢，观察硝苯吡啶在ＣＨＬ－３Ａ４ Ｓ９混合物（Ｓ９ｍｉｘ）中孵育前后的生物学活性变化来判断 ＣＨＬ－３Ａ４细胞的ＣＹＰ３Ａ４硝苯吡啶氧化酶活性。结果： Ｋ５６２ｒ细胞在含有硝苯吡啶（ ＮＩＦ）（ １２．５μｇ·Ｌ－１）的培养基中培养时，对阿霉素的ＩＣ５０值从不含 ＮＩＦ时的（６．４７± ０．６０）ｍｇ· Ｌ－１降至（０．８９±０．１５）ｍｇ· Ｌ－１（Ｐ＜０．０１）。Ｋ５６２ｒ细胞在含有分别经ＣＨＬ－３Ａ４、ＣＨＬ细胞的Ｓ９组分和灭活的ＣＨＬ－３Ａ４细胞的Ｓ９组分预处理的 ＮＩＦ（１２．５μｇ·Ｌ－１）的培养基中培养时，阿霉素对其的ＩＣ５０值分别为（６．１０±０．５０）ｍｇ· Ｌ－１、（０．３２±０．０９）ｍｇ·Ｌ－１和（０．３２±０．０４）ｍｇ·Ｌ－１。前者和后两者相比Ｐ＜０．０１。结论：实验室构建的表达人细胞色素Ｐ４５０ ３Ａ４同功酶转基因细胞系 ＣＨＬ－ ３Ａ４具有硝苯吡啶氧化酶活性，从而进一步确证?
OBJECTIVE: To detect the nifedipine oxidase activity of CHL-3A4 cells expressing human cytochrome P450 3A4 isoenzyme in our laboratory. Methods: Adriamycin (ADR) resistance in multidrug-resistant cells K562r was reversed by nifedipine, which in turn was specifically metabolized by CYP3A4. The effects of nifedipine on the growth of CHL-3A4 S9 mixture (S9mix ) Before and after incubation in biological activity changes to determine CHL-3A4 CYP3A4 nifedipine oxidase activity. Results: The IC50 values of doxorubicin in K562r cells cultured in medium containing nifedipine (NIF) (12.5μg · L-1) were (6.47 ± 0.60) mg · L-1 to (0.89 ± 0.15) mg · L-1 (P <0.01). K562r cells were cultured in medium containing NIF (12.5 μg · L-1) pre-treated with S9 fraction of CHL-3A4, CHL cells and S9 fraction of inactivated CHL-3A4 cells, respectively The IC50 values were (6.10 ± 0.50) mg · L-1, (0.32 ± 0.09) mg · L-1 and (0.32 ± 0.04) mg · L -1. The former and latter two were P <0.01. CONCLUSION: The laboratory-built human cytochrome P450 3A4 isozyme transgenic cell line CHL-3A4 has nifedipine oxidase activity, which confirms further.