RNAi沉默中期因子基因表达对胃癌细胞黏附与侵袭力的影响.docVIP

RNAi沉默中期因子基因表达对胃癌细胞黏附和侵袭力的影响 （作者:___________单位: ___________邮编: ___________） 作者：祁卫东 蒋鹏程 张恒 马圭 范钰 【摘要】 目的： 探讨中期因子(midkine，MK)基因小干扰RNA对胃癌细胞生物学行为的影响。方法： 培养人胃癌BGC823、HGC27、MGC803及SGC7901细胞株，以荧光实时定量PCR方法检测中期因子基因mRNA表达;筛选出中期因子表达最高者。采用中期因子基因小干扰RNA转染胃癌细胞株，分别以荧光实时定量PCR和免疫荧光方法观察中期因子基因mRNA和蛋白水平，然后以四唑蓝(MTT)比色法检测细胞黏附性，以Boyden方法检测癌细胞侵袭力。结果： 4株胃癌细胞中，中期因子均有不同程度的表达，以胃癌BGC823细胞最高;以MK siRNA转染胃癌BGC823细胞后，癌细胞中期因子基因mRNA和蛋白水平明显下降，且呈浓度依赖性;转染组细胞黏附数量明显下降，细胞侵袭力明显下降。结论： 中期因子基因在胃癌细胞黏附和侵袭中发挥着重要作用;以siRNA转染胃癌细胞，可抑制胃癌细胞黏附和侵袭能力。 【关键词】 胃癌; 中期因子; RNA干扰; 黏附; 侵袭 　[Abstract] Objective: To study the effects of midkine(MK) gene small interfering RNA(siRNA)on adhesion and invasion of human gastric cancer cell. Methods: Real time PCR was used to evaluate the MK mRNA expression of human gastric cancer cell lines BGC823,HGC27,MGC803 and SGC7901.The cell of MK highest expression was transfected with different dose of MK siRNA. The expression of MK mRNA and proteinwere were determined by realtime quantitative PCR and immumoflurescence method.The cell adhesion was evaluated by MTT assay, and invasion was exmined by Boyden chamber,respectively. Results: Cell line BGC823 showed the highest elevation of MK mRNA in four gastric cancer cell lines. The results from realtime quantitative PCR and immumoflurescence method showed that MK mRNA and protein reduced in timeand dosedependent manners(P0.01;P0.01).The adhesive, and invasive ability of BGC823 cell treated with MK siRNA decreased compared with control groups(P0.01, P0.01). Conclusion: MK gene might play an important role in adhesion and invasion of human gastric cancer cell. siRNA targeted MK could effectively inhibit adhesion,migration,and invasion of human gastric cancer cell. 　　[Key words] gastric carcinoma; midkine; RNA interference; adhesion; invasion 　　尽管胃癌的诊断和治疗已经有了长足的进步，但进展期胃癌的预后依然很差，5年生存率徘徊在20%～30%。己经证实，胃癌的发生发展是多基因改变的结果。胃癌中特殊基因的表达异常在不同的细胞功能中发挥了重要作用，如细胞黏附、信号传导、细胞分化、细胞增殖及转移。更好地了解胃癌发生发展的分子机制将有利于胃癌的诊治及预防。 　　许多研究表明生长因子不仅促进组织细胞的增殖，而且能诱导