ANTICANCER MECHANISMS OF LYCOPENE ACTION

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。前言：我们的目标是确定番茄红素的一些抗癌特性是否通过刺激缝隙连接通讯介导。今年，我们正在研究番茄红素是否会降低细胞的增殖，如果是的话，缝隙连接连接蛋白和mRNAs的产生是否会受到剂量依赖的影响。方法：我们筛选了6个细胞系：HS-578Bst，非癌乳腺；HS-578T，乳腺癌；HS-68，非癌皮肤成纤维细胞；DU-145，前列腺癌；ZR-75-1，乳腺癌；A549，肺癌；IMR-90，非癌肺。细胞暴露于10-10~10-5M的番茄红素剂量范围1-3天，然后进行电子计数。细胞生长至融合状态，番茄红素处理后，提取RNA，转化为cDNA，用聚合酶链式反应检测细胞间隙连接蛋白43基因的表达。结果：ZR-75-1细胞不能使用，HST-578T、HST-578Bst和IMR-90仍在筛选中。番茄红素对DU-145和HS-68细胞的增殖无明显影响。它们在整个剂量范围内表达连接蛋白43mRNAs。讨论：关于这些目标的研究仍然不完整，建筑工程延误了进度。结果表明，番茄红素对一些细胞系没有影响，因为它们的增殖和连接蛋白43的表达，因此将用于进一步的研究。这增加了我们的假设，即番茄红素的抗癌作用是它对缝隙连接通讯的影响，而不是对细胞增殖的影响。该项目将在本预算期剩余时间和下一个预算期继续进行这些研究。采用实时定量聚合酶链式反应(Real-time PCR)定量检测基因的表达，用ELISA法和Western blotting法检测细胞间隙连接蛋白的表达。将按计划增加更多的细胞系。我们将搬到一个新的实验室，并在这个预算期间建造。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction: Our goal is to determine whether some of the anticarcinogenic properties of lycopene are mediated through the stimulation gap junctional communication. This year we are investigating whether lycopene reduces cell proliferation and if so, whether production of gap junctional connexin proteins and mRNAs are affected in a dose-dependent fashion. Methods: We screened six human cell lines to date: Hs-578Bst, noncancerous breast; Hs-578T, breast carcinoma; Hs-68, noncancerous skin fibroblast; DU-145, prostate carcinoma; ZR-75-1, breast carcinoma; A549, lung carcinoma; and IMR-90, noncancerous lung. Cells were exposed to a dose-range of lycopene from 10-10 M to 10-5 M for 1-3 days and then counted electronically. The cells were grown to confluency, then treated with lycopene, then had their RNA separated, converted to cDNA and tested by PCR for expression of the connexin 43 gene. Results: The ZR-75-1 cells were not usable, the Hst-578T, Hst-578Bst and IMR-90 are still being screened. The DU-145 and Hs-68 cells have shown no significant changes to cell proliferation due to lycopene. They express the connexin 43 mRNAs across the dose-range. Discussion: Studies are still incomplete on these aims and building construction has delayed progress. The results show that several cell lines are not affected by lycopene because their proliferation and expression of the connexin 43 and therefore will be usable for further studies. This adds evidence to our hypothesis that the anticarcinogenic effect of lycopene is its effect on the gap junctional communication not cell proliferation. The project will continue with these studies for the remainder of the current budget period and into the next. We will add Real-time PCR to quantify the mRNA expression and use ELISA and western blotting to examine connexin protein expression. Additional cell lines will be added as planned. We will move to a new lab and building this budget period.