Carruba G, Webber M M, Bello-Deocampo D, Amodio R, Notarbartolo M, Deocampo N D, Trosko J E, Castagnetta L A
Institute of Oncology, University Medical School, and Experimental Oncology, Palermo Branch, National Cancer Institute of Genoa, Italy.
Anal Quant Cytol Histol. 1999 Feb;21(1):54-8.
To investigate gap-junctional intercellular communication (GJIC) in LNCaP and DU145 human prostate cancer cells.
Normal rat liver F344 (WB1) cells were used as positive controls. Functional GJIC was inspected using either the scrape-loading/dye transfer (SL/DT) method or fluorescence recovery after photobleaching (FRAP) analysis. In the former, GJIC activity was expressed as a measure of the extent of diffusion of Lucifer Yellow after cell monolayers were scraped using a surgical blade and exposed to dye for a few minutes at room temperature. In the latter, cells were incubated for 15 minutes at 37 degrees C with 5,6-carboxyfluorescein diacetate dye and the dye transfer visualized by photobleaching individual cells with a 488-nm laser and monitoring the recovery of fluorescence using a laser cytometer.
The preliminary results obtained indicate that neither LNCaP nor DU145 cells have functional GJIC, while, as expected, WB1 cells show unimpaired GJIC activity. Equivalent results were consistently obtained using either SL/DT or the FRAP approach. However, using FRAP analysis, DU145 cells only showed weak recovery of fluorescence after a total observation interval of 15 minutes.
The present data, though preliminary, suggest that disruption of GJIC may play a role in development of malignancy in the human prostate.
研究LNCaP和DU145人前列腺癌细胞中的间隙连接细胞间通讯（GJIC）。
正常大鼠肝脏F344（WB1）细胞用作阳性对照。使用刮擦加载/染料转移（SL/DT）方法或光漂白后荧光恢复（FRAP）分析来检测功能性GJIC。在前者中，GJIC活性通过使用手术刀刮擦细胞单层并在室温下暴露于染料几分钟后，以荧光素黄扩散程度来衡量。在后者中，细胞在37℃下与5,6-羧基荧光素二乙酸酯染料孵育15分钟，通过用488-nm激光对单个细胞进行光漂白并用激光细胞仪监测荧光恢复来观察染料转移。
获得的初步结果表明，LNCaP细胞和DU145细胞均无功能性GJIC，而正如预期的那样，WB1细胞显示出未受损的GJIC活性。使用SL/DT或FRAP方法均一致获得了相同的结果。然而，使用FRAP分析，DU145细胞在总共15分钟的观察间隔后仅显示出微弱的荧光恢复。
目前的数据虽然是初步的，但表明GJIC的破坏可能在人类前列腺恶性肿瘤的发生中起作用。
