SiRNA沉默Notch2表达对Lewis肺癌细胞增殖和周期的影响

来源 :现代生物医学进展 | 被引量 : 0次 | 上传用户:ericshen81
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
目的:采用RNA干扰技术下调Notch2基因在Lewis肺癌细胞(LLC)的表达,探讨Notch2表达下调对LLC增殖和周期的影响。方法:通过脂质体2000将Notch2-siRNA和对照con-siRNA分别转染LLC后,通过PCR,RT-PCR检测Notch2的表达情况;通过MTT、软琼脂克隆形成实验检测LLC的生长情况;并使用流式细胞技术检测细胞周期。结果:SiRNA转染LLC48小时后,PCR和RT-PCR检测结果显示Notch2在实验组的表达受到显著抑制;MTT实验显示干扰Notch2的表达显著抑制了LLC的增殖(P<0.001);软琼脂克隆形成实验表明干扰Notch2的表达将影响LLC的克隆形成;细胞周期实验进一步证实干扰Notch2的表达使处于活跃增殖期(S期)的LLC数量明显减少。结论:Notch2在LLC中的表达发挥重要的促癌作用,干扰Notch2的表达能显著抑制LLC的分裂增殖和克隆形成,从而抑制肿瘤的生长。 OBJECTIVE: To investigate the effect of down-regulation of Notch2 on the proliferation and the cycle of LLC by down-regulating the expression of Notch2 gene in Lewis lung carcinoma (LLC) cells by RNAi technique. METHODS: Notch2-siRNA and control con-siRNA were transfected into LLC by Lipofectamine 2000 respectively. The expression of Notch2 was detected by PCR and RT-PCR. The growth of LLC was detected by MTT and soft agar colony formation assay. Flow Cytometry detects cell cycle. Results: After48h siRNA transfection, the expression of Notch2 in the experimental group was significantly inhibited by PCR and RT-PCR. The MTT assay showed that the expression of Notch2inhibited the proliferation of LLC significantly (P <0.001) Experiments show that the interference of Notch2 expression will affect the formation of LLC clones; cell cycle experiments further confirmed that the interference of Notch2 expression in the active proliferative phase (S phase) significantly reduced the number of LLC. Conclusion: The expression of Notch2 in LLC plays an important role in carcinogenesis. Interference of Notch2 expression significantly inhibits the growth and proliferation of LLC cells.
其他文献
番茄斑萎病毒属病毒是一类重要的经济作物病毒,在世界范围内广泛分布,根据目前的报道,在南美、东南亚和南亚为集中分布区。Tospovirus属于布尼亚病毒科,该科病毒共5个属,其中4个
类风湿关节炎(RA)是一种以关节及其周围组织炎症为特征,以慢性、多发性、对称性关节肿痛为主要表现的常见全身性自身免疫疾病。中药伸筋草具有祛风除湿,舒筋活络之功效是临床
会议
提起保镖,再没有比惠特尼.休斯顿主演的Bodyguard一片里凯文.科斯特纳饰演的保镖给人留下的印象更为深刻了,那欲言又止的爱情描绘出保镖与受托人之间不可逾越的职业界限。保
A thermodynamic database on the phase equilibria of the uranium base alloys,including U-X(X:Al,Co,Cr,Cu,Ga,Mg,Mn,Mo,Nb,Th,W)binary system has been developed by the Calcul
会议
The composition range of amorphous forming for Al-Fe-Zr system has been determined by mechanical alloying of mixture of elemental powders.The phases of as-milled
会议
当贵为巴拿马总统、国防军总司令时,曼努埃尔·安东尼奥·诺列加绝不会想到,美国为逮捕他一个人竟发动了一场战争!从权力巅峰坠落的路,是那般的戏剧性。自1989年被赶下总统宝
The influence of C and Al content on phase transformation temperatures, i.e. the Al and A3 of Fe-rich alloys is investigated by dilatometric analysis. With the
会议
The models of phase diagram calculation of semiconductor thin films with different substrates were proposed by considering the contributions of strain energy,sel
会议
Serials of DyCo2-xGax alloys with x from O to 2 were prepared by arc melting and investigated by X-ray powder diffraction analysis.Four ternary compounds,DyCo1.33
会议
The nature of the second law of thermodynamics is the irreversibility of time for the development of macroscopic systems,i.e.,“arrow of time”.However,Carnot theore
会议