目的:探讨二肽基肽酶(DDPIV)对前列腺癌细胞IE8侵袭、转移的影响。方法:用亚克隆技术构建DDPⅣ基因反义真核表达质粒,脂质体法将其分别转入高转移潜能的人前列腺癌细胞1E8,并应用脂质体介导的基因转染技术,将重组质粒导入1E8细胞中,应用细胞增殖抑制试验(CCK-8)、蛋白印迹和体外侵袭实验等方法观察了IE8细胞转染前后,细胞生长、DDPⅣ蛋白表这以及细胞体外侵袭能力等指标的变化。结果:成功构建了反义DDPIV真核表达载体;将其转染入1E8细胞36h、48h后,反义DDPⅣ基因对前列腺癌细胞的侵袭和转移有促进作用;DDPⅣ基因表达下调能使1E8细胞的体内外侵袭能力增强。结论:DDPⅣ是前列腺癌的一个抑癌基因,在前列癌的侵袭转移过程中发挥着重要作用,下调DDPⅣ基因表达能明显促进前列腺癌细胞的侵袭和转移能力。提示DDPⅣ可作为前列腺癌抗侵袭治疗的分子靶点。
Objective: To evaluate the effect of Dipeptidyl Peptidase IV on the invasion and metastasis of prostatic carcinoma cell line PC-3M-1E8. Methods: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify dipeptidyl peptidase Ⅳ (DDPⅣ) cDNA fragments. RT-PCR products were cloned into plasmid pEGFP-C1 in the proper orientation for expressing antisense RNA, and the recombinant pEGFP-CI/DDP Ⅳ was transfected into 1 E8 cells. We employed the CCK-8 assay to detect changes in cell proliferation. DDP IV protein expression and invasiveness were detected by Western blot and wound healing assay, respectively. Results: The eukaryotic expression vector pEGFP-C1/DDPIV designed to produce antisense DDPⅣ RNA was constructed successfully. At 36 hours and 48 hours after transfection with pEGFP-CI/DDPⅣ, the proliferation of 1E8 cells was significantly higher than that of untransfected cells. Compared with control cells, the pEGFP-CI/DDP Ⅳ-transfected cells displayed decreased expression of endogenous DDP Ⅳ protein. Tumor cell motility and invasive potential were enhanced in pEGFP-Cl/DDPⅣ-transfected cells. Conclusion: DDPⅣ is a tumor suppressor gene that inhibits metastasis of prostatic cancer. Proliferation and invasive potential of 1E8 cells were enhanced by antisense DDPⅣ, suggesting that DDPⅣ may be useful in gene therapy for prostatic carcinoma.