应用刻痕标记／荧光扩散试验检测抗促癌物的初步研究

应用刻痕标记荧光扩散试验,可以观察到促癌物TPA抑制缝隙连接介导的细胞间通讯(GJIC)。本文探讨用荧光扩散试验检测抗促癌物的可能性。经小鼠皮肤乳头状瘤诱发试验检查的3种已知抗促癌物(姜黄素,迷迭香抗氧化剂,和绿茶提取物),均能明显阻断TPA抑制BALB/C3T3细胞间的荧光扩散作用。用文献中报道的另一种筛选抗促癌物的方法,找到12种中草药抑制TPA诱导小鼠耳皮肤水肿,其中9种在本荧光扩散试验中可以明显拮抗TPA的作用,另外6种中草药在这两种试验中均为阴性。本研究初步表明,刻痕标记/荧光扩散试验有可能用于抗促癌物的快速检测。

THEUSE OF SCRAPE- LOADING/ DYE TRANSFER ASSAY FOR DETECTION OF ANTIPROMOTERS

Tumor promoter TPA inhibit gap junctional mediated intercellular communication (GJIC), which can be detected by a simple scrape loading/dye transfer assay. The potential of using this assay for the detection of antipromoters was investigated. Three antipromoters (curcumine. rosemary antiox idant, and green tea extract) which have already been indentified by mouse skin papilloma tests strong ly removed TPA caused GJIC inhibition in this assay with BALB/c 3T3 cells. This assay was further used for testing the potential antipromotion effect of a group of Chinese herbal medicines which have been identified by the inhibition of TPA-induced mouse ear edema, a proposed assay for pre-screening antipromoters. Nine out of the twelve Chinese herbal medicines are coincidential in both assays as posi live, other six Chinese herbal medicines are negative in both assays. This preliminary results indicated that scrape-loading/dye transfer assay may be useful as a rapid screening method for the detection of antipromoter.