HELQ对骨肉瘤细胞增殖、侵袭、迁移的影响及机制

摘要

Objective To investigate the effects and mechanism of HELQ (helicase,POLQ-like) on the proliferation,invasion,and migration of osteosarcoma cells.Methods The human osteosarcoma cell lines (U2-OS and 143B) were transfected with Lv-HELQ,Lv-Negative,Lv-shHELQ,respectively,and were divided into three groups:Lv-HELQ group,NC group,and Lv-shHELQ group.The expression of HELQ protein at 48 h was detected by Western blotting.The results showed that the expression of HELQ in these two cell lines of the Lv-HELQ group was significantly higher,while the expression of HELQ in these two cell lines of the Lv-shHELQ group was significantly lower than that in the control group (all P ＜ 0.05),which indicated that the transfection was successful.At 48 h after transfection,the proliferation,migration,and invasion abilities of these two cell lines,and DNA damage levels were investigated by CCK8,Wound healing,Transwell invasion,and comet assay,respectively.Results The OD values of these two cell lines were significantly higher in the Lv-HELQ group but significantly lower in the Lv-shHELQ group as compared with those of the NC group (P ＜ 0.05 or P ＜ 0.01).The migration rate,transmembrane cells,and percentage of DNA in tail DNA of these two cell lines were significantly lower in the Lv-HELQ group but significantly higher in the Lv-shHELQ group as compared with those of the NC group (all P ＜ 0.05).Conclusion HELQ can inhibit the proliferation,invasive,and migration of osteosarcoma cells by repairing the DNA damage.%目的 探讨POLQ样蛋白解旋酶(HELQ)对骨肉瘤细胞增殖、侵袭、迁移的影响及作用机制.方法 将体外培养的人源骨肉瘤细胞系U2-OS、143B随机分为Lv-HELQ组、NC组、Lv-shHELQ组,分别转染Lv-HELQ、Lv-Negative、Lv-shHELQ慢病毒载体.转染48 h时,采用Western blotting法检测HELQ蛋白表达,结果显示两种细胞Lv-HELQ组HELQ蛋白相对表达量均高于NC组,Lv-shHELQ组均低于NC组,组间比较P均＜0.05,提示HELQ转染成功.三组(两种细胞)转染48 h时,采用CCK8法检测细胞增殖情况(光密度值),采用Wound healing法检测细胞迁移率,采用Transwell invasion法检测细胞穿膜细胞数(侵袭能力),采用彗星实验检测彗星尾部DNA百分比.结果 两种细胞Lv-HELQ组光密度值均高于NC组,Lv-shHELQ组光密度值均低于NC组,组间比较P＜0.05或＜0.01.两种细胞Lv-HELQ组细胞迁移率、穿膜细胞数及均彗星尾部DNA百分比均低于NC组,Lv-shHELQ组均高于NC组,组间比较P均＜0.05.结论 HELQ可抑制人骨肉瘤细胞增殖、侵袭、转移,修复DNA损伤可能是其作用机制.