沉默circ9119调控miR-126抑制肝癌细胞的增殖、侵袭和自噬

Silencing circ9119 inhibits proliferation, invasion and autophagy of hepatocellular carcinoma cells by regulating miR-126

目的:探讨环状RNA-9119(circular-RNA-9119,circ9119)在肝癌(hepatocellular carcinoma, HCC)中的作用及其作用机制。方法:qRT-PCR检测收集的组织样本和购买的细胞系中circ9119、miR-126的表达,并对细胞进行筛选。借助荧光原位杂交(FISH)实验、RNA结合蛋白免疫共沉淀(RIP)实验与双荧光素酶报告实验对circ9119和miR-126的关系进行明确。凋亡相关因子和自噬标记物借助qRT-PCR、Western blot测定。CCK8、Transwell、流式细胞术分别测定HCC细胞的增殖、侵袭和凋亡。裸鼠成瘤实验观测肿瘤生长状况。结果:HCC组织和细胞中,circ9119的表达上调、miR-126的表达下调(均P<0.05)。circ9119能够作为miR-126的ceRNA对其进行调控。沉默circ9119有利于抑制HCC细胞的增殖、侵袭和自噬,促进细胞的凋亡,且能够抑制小鼠体内肿瘤的生长(均P<0.05)。而过表达circ9119则变化与之相反。上调miR-126的表达与沉默circ9119中细胞变化趋势一致。上调miR-126的作用能够被过表达circ9119所逆转。结论:circ9119作为促癌因子负调控miR-126在HCC中发挥作用。

Objective:To identify the role and mechanism of circular-RNA-9119(circ9119) in hepatocellular carcinoma(HCC).Methods:The expression of circ9119 and miR-126 in the collected tissue samples and purchased cell lines were detected by qRT-PCR,and cells were screened.FISH assay, RIP assay and luciferase reporter assay were peroformed to confirm the relationship between circ9119 and miR-126.Apoptosis related factors and autophagy markers were tested by qRT-PCR and Western blot.The proliferation, invasion and apoptosis of HCC cells were measured by CCK8 assay, Transwell assay and flow cytometry assay respectively.Tumor growth was observed in nude mice.Results:The expression of circ9119 was increased in HCC tissue and cells, while the expression of miR-126 was decreased in HCC tissues and cell(P<0.05).circ9119 could regulate miR-126 by functioning as a ceRNA.Silencing of miR-126 could inhibit the proliferation, invasion and autophagy, promote the apotposis of HCC cells, and restrain the growth of tumor in mice(P<0.05). However, the effects of overexpression of circ9119 was opposite to that. The changes in cells of up-regulating miR-126 expression and down-regulating circ9119 expres-sion were in the same trend. The effects of up-regulating miR-126 expression could be reversed by overexpressionof circ9119.Conclusion:Together,our findings suggested that circ9119 functions as an oncogene in HCC by regula-ting miR-126 negatively.