Analysis And Funtion Studies Of Unique Segments Of PC-1 And Primarily Research On Anti-cancer Effect Of Kenaf Seeds Oil

PC-1 is a prostate cancer related gene that locates at chromosome 8q21.1,a locus most frequently amplified in prostate cancer. The PC-1gene product belongs to TPD52 family and its first 46 amino acids in N-terminal make it unique to other members. Even induced by androgen,PC-1 is expressed in all kinds of prostate cancer cell lines .And with the clinical stage and malignancy increase ,the expression of PC-1 is up-regulated. Previously studies have shown that PC-1 has the characteristics of transcription factors and promotes the proliferation and androgen-independent cells growth. CLAs , essential components of kenaf seeds oil,which derviatived from essential linoleic acid,are important to human physiological functions and widely exist in animals and plants.A pile of researches from home and abroad have showed that CLAs are capable of inhibit a various of tumors. Accordingly,we use a series of experiments to study the specific segment of PC-1 and anti-cancer effects of CLAs, and the following results were obtained:1)By using overlapping PCR and other molecular cloning technology , deletants and mutants of PC-1 that fused to green fluorescent protein have been constructed. Having transfected these vectors into LNCaP cell ,we found that the minmus size of cytoplasmic location of PC-1 may ranges from 111～114 amino residues. Meanwhile, even 103～114 amino residues are important for PC-1's subcellular localization, but they are not nuclear exporting sequence as previously predict.2) We successfully construct the adenovirus vectors of pAd-EGFP,pAd-EGFP-PC11-46,pAd-EGFP-PC-147-224 and pAd-EGFP-PC-1 by using homologous recombination technique.And these vectors provided prerequisites to study the function of PC-1 fragments.3)While colony-formation assay proved that CLA inhibits cancer cells growth in vitro, westernblotting assay show the ability of CLA to down-regulate Akt's phosphorylation level.In this study, we found a sequence that markedly impact PC-1's subcellular localization and successfully constructed the adenovirus vectors for function study of PC-1 segments . At the same time, we found that CLA inhibited a various of cancer cells growth in vitro and down-regulate AKT's phosphorylation level in C4-2 cell line ,which suggesting its potential value in medical and health food .