ANTICANCER MECHANISMS OF LYCOPENE ACTION

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们的目的是确定番茄红素的一些抗癌特性是否通过刺激间隙连接通讯介导。今年，我们正在研究番茄红素是否会减少细胞增殖，如果是这样，间隙连接蛋白和mrna的产生是否会以剂量依赖性的方式受到影响。方法：迄今为止，我们筛选了6种人类细胞系：Hs-578Bst，非癌性乳腺癌；Hs-578T，乳腺癌；Hs-68，非癌性皮肤成纤维细胞；DU-145，前列腺癌；ZR-75-1，乳腺癌；A549，肺癌；和IMR-90，非癌性肺。将细胞暴露于10-10 M至10-5 M剂量范围的番茄红素中1-3天，然后进行电子计数。将细胞培养至融合后，用番茄红素处理，分离RNA，转化为cDNA，用PCR检测连接蛋白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。茄红素对DU-145和Hs-68细胞增殖无明显影响。它们在整个剂量范围内表达连接蛋白43 mrna。讨论：关于这些目标的研究仍然不完整，建筑的建设推迟了进展。结果表明，一些细胞系不受番茄红素的影响，因为它们的增殖和连接蛋白43的表达，因此可用于进一步的研究。这为我们的假设增加了证据，即番茄红素的抗癌作用是对间隙连接通讯的影响，而不是对细胞增殖的影响。本项目将在本预算期的剩余时间和下一个预算期继续进行这些研究。我们将采用Real-time PCR定量mRNA表达，ELISA和western blotting检测连接蛋白表达。其他细胞系将按计划添加。在这个预算期内，我们将搬到一个新的实验室。

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.