沉默TRIM29对骨肉瘤细胞增殖、凋亡及侵袭的影响

目的 探讨小干扰RNA（siRNA）靶向三联基序蛋白29（TRIM29）对骨肉瘤细胞增殖、凋亡及侵袭能力的影响。方法 采用实时荧光定量PCR（QPCR）法检测骨肉瘤细胞株（MG63和U2OS）的TRIM29水平，采用脂质体法将两条靶向TRIM29的siRNA片段（siTRIM29-1和siTRIM29-2）高效转染至MG63和U2OS细胞，经QPCR法检测后选取抑制效率最高的siRNA进行后续实验，并设转染siControl的MG63和U2OS细胞为对照；分别采用MTT、流式细胞术及Transwell 法检测转染后MG63和U2OS细胞的增殖、凋亡及侵袭情况。结果 QPCR检测结果显示MG63和U2OS细胞的TRIM29 mRNA水平较正常成骨细胞株hFOB1-19均升高（P＜0.05）；MG63和U2OS细胞转染siTRIM29-1和siTRIM29-2片段72 h后的TRIM29 mRNA水平均低于转染siControl的细胞，差异均有统计学意义（P＜0.05），同时选取沉默率较高的siTRIM29-1进行后续实验；与转染siControl的MG63和U2OS细胞相比，两种细胞转染siTRIM29-1后的存活率及穿膜细胞数目减少，凋亡率升高，差异均有统计学意义（P＜0.05）。结论 骨肉瘤细胞中TRIM29高表达，且靶向沉默TRIM29可抑制骨肉瘤细胞增殖及侵袭并诱导细胞凋亡，对骨肉瘤靶向治疗有一定参考价值。

Silencing effect of TRIM29 on proliferation,apoptosis and invasion of osteosarcoma cells

Objective To investigate the effect of small interfering RNA (siRNA) targeting tripartite motif-containing 29 (TRIM29) on the proliferation,apoptosis and invasion of osteosarcoma cells.Methods The levels of TRIM29 in osteosarcoma cell lines (MG63 and U2OS) were detected by fluorescence real-time quantitative PCR (QPCR) assay.Two siRNA fragments (siTRIM29-1 and siTRIM29-2) were efficiently transfected into MG63 and U2OS cells by lipofectamine method,and MG63 and U2OS cells transfected with siControl were set as control.The highest inhibitive efficiency of siRNA was selected for subsequent functional study.The proliferation,apoptosis and invasion of MG63 and U2OS cells after transfection were evaluated by MTT,flow cytometry and Transwell assay.Results The results of QPCR test showed that TRIM29 mRNA of MG63 and U2OS cells were significantly higher than those of normal osteoblast cell line hFOB1.19 (P＜0.05).Transfection of TRIM29 siRNA could reduce the TRIM29 level of MG63 and U2OS cells at 72 h.Compared with the cells transfected with siControl,there were decreased survival rates and transmembrane cell number,but increased apoptotic rates in transfection group with statistical significance (P＜0.05).Conclusion TRIM29 is highly expressed in osteosarcoma cells,and silencing of TRIM29 can inhibit the proliferation and invasion of osteosarcoma cells and induce cell apoptosis,which can be used as reference in the treatment of osteosarcoma.