LINC00261/miR-182-5p/PFN1对乙肝病毒相关性肝细胞癌HepG2.2.5细胞放疗抵抗的影响

Effect of LINC00261/miR-182-5p/PFN1 on radiotherapy resistance of hepatitis B virus-related hepatocellular carcinoma cell HepG2.2.15

[目的]探讨LINC00261调控miR-182-5p/PFN1轴对乙型肝炎病毒相关性肝细胞癌HepG2.2.15细胞放疗抵抗的作用机制。[方法]用1 Gy的X射线处理HepG2.2.15细胞得放疗抵抗细胞HepG2.2.15/R,RT-qPCR检测LINC00261和miR-182-5p表达,Western Blot检测PFN1的表达,CCK-8检测细胞增殖活力,流式细胞术检测细胞凋亡率,彗星实验检测DNA损伤情况;双荧光素酶报告基因实验验证miR-182-5p和LINC00261或PFN1的靶向关系。[结果]LINC00261在HepG2.2.15细胞中低表达(P<0.01),且在其放疗抵抗细胞HepG2.2.15/R中表达更低(P<0.01)。过表达LINC00261抑制HepG2.2.15/R细胞增殖(P<0.01),诱导细胞凋亡(P<0.05)和DNA双链断裂(P<0.01);6 Gy X射线处理可上调过表达LINC00261对HepG2.2.15/R细胞凋亡(P<0.05)和DNA损伤的促进作用(P<0.01)。双荧光素酶报告基因实验证实miR-182-5p与LINC00261或PFN1的靶向关系。过表达LINC00261或过表达PFN1可下调过表达miR-182-5p对HepG2.2.15/R细胞增殖的促进作用(P<0.05)以及对凋亡和DNA损伤的抑制作用(P<0.05)。[结论]过表达LINC00261靶向下调miR-182-5p,促进PFN1表达,促进HepG2.2.15细胞放疗抵抗。

[Objective] To explore the mechanism of LINC00261 regulating miR-182-5 p/PFN1 axis on radiation resistance of hepatitis B virus-related hepatocellular carcinoma HepG2.2.15 cells.[Method] HepG2.2.15 cells were treated with 1 Gy of X-ray to obtain radiotherapy resistant cells HepG2.2.15/R,and the expressions of LINC00261 and miR-182-5 p was detected by RT-qPCR,the expression of PFN1 was detected by Western Blot,the proliferation of cells was detected by CCK-8,the apoptosis rate was detected by flow cytometry,and DNA damage was detected by comet assay.Double luciferase reporter gene experiment verified the targeting relationship between miR-182-5 p and LINC00261 or PFN1.[Result] LINC00261 was poorly expressed in HepG2.2.15 cells(P<0.01),and even lower in its radiotherapy-resistant cell HepG2.2.15/R(P<0.01).Overexpression of LINC00261 inhibited the proliferation of HepG2.2.15/R cells(P<0.01),induced apoptosis(P<0.05) and DNA double-strand breaks(P<0.01).6 Gy X-ray treatment up-regulated the effect of overexpression of LINC00261 on HepG2.2.15/R cell apoptosis(P<0.05) and DNA damage(P<0.01).The dual luciferase reporter gene experiment confirmed the targeting relationship between miR-182-5 p and LINC00261 or PFN1.Overexpression of LINC00261 or overexpression of PFN1 could down-regulate the promotion effect of overexpression of miR-182-5 p on proliferation of HepG2.2.15/R cells(P<0.05) and inhibition of apoptosis and DNA damage(P<0.05).[Conclusion] Overexpression of LINC00261 down-regulated miR-182-5 p,promoted PFN1 expression and promoted radiation resistance of HepG2.2.15 cells.