文档介绍：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 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 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(;).The adhesive, and invasive ability of BGC823 cell treated with MK siRNA pared with control groups(, ). 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 in
terference; 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,而且能诱导组织细胞的恶性转化,在肿瘤的发展中发挥了