CAJ | 학술논문

目的构建保罗样激酶-1(PLK1)基因小干扰RNA(siRNA),并观察其对甲状腺癌细胞增殖和凋亡的影响.方法设计合成5个PLK1 siRNA(S1、S2、S3、S4、S5),转染人甲状腺癌ARO细胞后,采用实时定量RT-PCR方法筛选下调PLK1 mRNA效果最好的siRNA;然后以该siRNA转染ARO细胞后,分别以实时定量RT-PCR和Western blot检测各组细胞PLK1基因表达情况;以MTT法检测对各组细胞增殖的影响;分别以caspase-3活性和TUNEL方法明确甲状腺癌细胞凋亡情况.结果 5个siRNA均明显下调PLK1 mRNA水平,以S4效果最好,抑制率达91.5%.MTr结果显示,S4 siRNA转染对甲状腺癌细胞增殖均有明显的抑制作用,且呈浓度和时间依赖性(P<0.05).与对照组比较,S4 siRNA转染组癌细胞caspase-3活性明显升高,且呈浓度和时间依赖性(r=0.919;r=0.957);TUNEL结果显示,S4 siRNA转染组出现明显的凋亡现象,且呈浓度依赖性(r=0.932).结论 PLK1基因在甲状腺未分化癌细胞增殖中具有重要的调控作用.PLK1 siRNA转染可明显抑制癌细胞增殖,诱导凋亡是其重要机制之一.
목적 구건보라양격매-1(PLK1)기인소간우RNA(siRNA),병관찰기대갑상선암세포증식화조망적영향.방법 설계합성5개PLK1 siRNA(S1、S2、S3、S4、S5),전염인갑상선암ARO세포후,채용실시정량RT-PCR방법사선하조PLK1 mRNA효과최호적siRNA;연후이해siRNA전염ARO세포후,분별이실시정량RT-PCR화Western blot검측각조세포PLK1기인표체정황;이MTT법검측대각조세포증식적영향;분별이caspase-3활성화TUNEL방법명학갑상선암세포조망정황.결과 5개siRNA균명현하조PLK1 mRNA수평,이S4효과최호,억제솔체91.5%.MTr결과현시,S4 siRNA전염대갑상선암세포증식균유명현적억제작용,차정농도화시간의뢰성(P<0.05).여대조조비교,S4 siRNA전염조암세포caspase-3활성명현승고,차정농도화시간의뢰성(r=0.919;r=0.957);TUNEL결과현시,S4 siRNA전염조출현명현적조망현상,차정농도의뢰성(r=0.932).결론 PLK1기인재갑상선미분화암세포증식중구유중요적조공작용.PLK1 siRNA전염가명현억제암세포증식,유도조망시기중요궤제지일.
Objective To study effects of polo-like kinase-1(PLK1)small interfering RNA(siRNA)on proliferation and apoptosis of undifferentiated human thyroid cancer cells.Methods 5 PLK1 siRNA(S1,S2,S3,S4 and S5)were constructed and used to transfect human thyroid cancer cell line ARO.RT-PCR was employed to pick out the most effective siRNA,which was then used to transfect ARO cell.RT-PCR and western blot were used to detect PLK1 expression in thyroid cancer cells,which were divided into different groups.MTT assay was performed to examine the effects of PLK1 siRNA on thyroid cancer cells in all groups.Apoptosis of thyroid cancer cells was observed by caspase-3 activity and TUNEL.Results All the 5 siRNA down-regulated PLK1 mRNA expression.among which S4 showed the best effect.S4 transfection could obviously inhibit proliferation of thyroid cancer cells in dose and time dependent manner.Compared with control groups,caspase-3 activity of cancer cells in s4 transfeeted group increased significantly.The effect of S4 transfection was dose and time dependent.TUNEL results showed apoptosis of cancer cells transfected by S4 siRNA was obvious and apoptosis of cells was dose-dependent.Conclusions PLK1 may play an important role in proliferation of undifferentiated thyroid carcinoma.PLK1 siRNA transfection can inhibit proliferation of throid cancer cell through apoptosis induction.