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.
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