小干扰RNA抑制骨肉瘤细胞组织蛋白酶K表达及生长与侵袭的研究

Inhibitory effect of small interfering RNA on Cathepsin K expression and cell growth and invasion ability of osteosarcoma cells

目的 观察人小干扰RNA （siRNA）沉默入骨肉瘤细胞株MG63组织蛋白酶K（Cathepsin K）对增殖和侵袭影响.方法 利用脂质体将Cathepsin K siRNA转染MG63,通过反转录-聚合酶链反应（RT-PCR）与Western blot分别检测Cathepsin K mRNA及蛋白质表达,增殖与迁移实验评估沉默Cathepsin K后MG63增殖与侵袭改变.结果 siRNA能有效地抑制Cathepsin KmRNA及蛋白质表达（0.05±0.16、4.00±0.08）,与对照组（1.10 ±0.06、1.00±0.21）比较,差异有统计学意义（P＜0.01）;沉默Cathepsin K细胞增殖（100.00±0.09）明显低于对照组（40.00±0.13,P＜0.01）;Cathepsin K siRNA组迁移细胞数[（45.0±4.9）个]明显低于对照组[（112.0±8.0）个,P＜0.01].结论 siRNA可有效抑制Cathepsin K表达,降低MG63增殖与侵袭能力.

Objective To explore the inhibitory effect of small interfering RNA （siRNA） on Cathepsin K expression and growth and invasion ability of osteosarcoma cells.Methods A Cathepsin K mRNA-targeting siRNA was transfected into MG63 cells with lipidosme.Reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting were used to detect the mRNA and protein expression of Cathepsin K respectively.Methyl thiazol tetrazolium （MTT） and Transwell assays were applied to measure the proliferation and invasion of osteosarcoma cells.Results The mRNA and protein expression levels of Cathepsin K in the cells transfected with Cathepsin K siRNA （0.05 ±0.16 and 4.00 ±0.08） were lower than in the control group （1.10 ±0.06 and 1.00 ±0.21,P 〈0.01）.The proliferation index in Cathepsin K-silenced cells （100.00 ±0.09） was lower than that in control group （40.00 ±0.13,P 〈0.01）.The number of invasion cells in Cathepsin K siRNA group （45.0 ±4.9） was significantly less han in control group （112.0 ± 8.0,P 〈 0.01）.Conclusion siRNA can effectively silence the expression of Cathepsin K and inhibit the proliferation and invasion of osteosarcoma cells.