HELQ对骨肉瘤细胞增殖、侵袭、迁移的影响及机制

发布时间：2019-01-07 13:48

【摘要】：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-sh HELQ组,分别转染Lv-HELQ、Lv-Negative、Lv-sh HELQ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-sh HELQ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-sh HELQ组光密度值均低于NC组,组间比较P0.05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-sh HELQ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。
[Abstract]:Objective to investigate the effect and mechanism of POLQ like protein helicase (HELQ) on proliferation, invasion and migration of osteosarcoma cells. Methods Human osteosarcoma cell line U2-OS-143B was randomly divided into Lv-HELQ group, NC group and Lv-sh HELQ group. The cells were transfected with Lv-HELQ,Lv-Negative,Lv-sh HELQ lentivirus vector. At 48 h after transfection, the expression of HELQ protein was detected by Western blotting method. The results showed that the relative expression of HELQ protein in Lv-HELQ group was higher than that in NC group, and that in Lv-sh HELQ group was lower than that in NC group (P 0.05), indicating that HELQ transfection was successful. After 48 h transfection of three groups of cells (two kinds of cells), the cell proliferation (optical density) was detected by CCK8 assay, the cell migration rate was measured by Wound healing assay, and the number of cell transmembrane cells (invasiveness) was measured by Transwell invasion method. Comet assay was used to detect the percentage of comet tail DNA. Results the optical density of Lv-HELQ group was higher than that of NC group, and that of Lv-sh HELQ group was lower than that of NC group. The cell mobility, the number of transmembrane cells and the percentage of DNA in the tail of comet were lower in Lv-HELQ group than in NC group, but higher in Lv-sh HELQ group than in NC group (P 0.05). Conclusion HELQ can inhibit the proliferation, invasion and metastasis of human osteosarcoma cells and repair the damage of DNA.
【作者单位】：井冈山大学附属医院;南昌大学第一附属医院;深圳市南山区人民医院;
【分类号】：R738.1

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