沉默lncRNA UCA1通过上调miR-873-5p表达对胶质瘤细胞放射敏感性影响

目的 探讨lncRNA UCA1通过调控miR-873-5p表达对体外培养的胶质瘤SHG-44、U87、U251细胞放射敏感性的影响。方法 采用克隆形成实验检测X线照射0、2、4、6、8Gy SHG-44、U87、U251细胞存活情况,qRT-PCR检测SHG-44、U87、U251细胞中UCA1基因表达。以放射抵抗的U87、U251细胞为研究对象,沉默UCA1表达、过表达miR-873-5p后用克隆形成实验及流式细胞术检测细胞存活分数和细胞凋亡率。双荧光素酶报告基因实验和qRT-PCR检测验证UCA1和miR-873-5p的靶向关系。结果 UCA1在U87、U251细胞中表达上调。沉默UCA1或过表达miR-873-5p可抑制U87、U251细胞存活,并促进细胞凋亡。miR-873-5p是UCA1靶基因,UCA1可负性调控miR-873-5p1的表达。抑制miR-873-5p表达可逆转沉默UCA1对胶质瘤细胞放射敏感性的影响。沉默UCA1增加射线对胶质瘤细胞U251移植瘤的抑制作用。结论 沉默UCA1通过上调miR-873-5p表达,抑制胶质瘤细胞存活并促进其凋亡,从而提高胶质瘤细胞放射敏感性。

Silencing lncRNA UCA1 affects radiosensitivity of glioma cells by up-regulating miR-873-5p expression

Objective To investigate the effect of lncRNA UCA1 on the radiosensitivity of in vitro cultured glioma cell lines SHG-44, U87 and U251 by regulating the miR-873-5p expression. Methods The survival of glioma cells SHG-44, U87 and U251 treated with different radiation intensities (0, 2, 4, 6 and 8Gy) was detected by colony formation assay. The expression levels of UCA1 in glioma cells SHG-44, U87 and U251 were measured by qRT-PCR. The radiation-resistant glioma cells U87 and U251 were selected for subsequent study. After silencing UCA1 expression and/or over-expressing miR-873-5p, the cell survival rate was detected by colony formation assay, and the cell apoptosis rate was determined by flow cytometry. The dual luciferase reporter gene assay and qRT-PCR were employed to verify the targeting relationship between UCA1 and miR-873-5p. Results UCA1 was up-regulated in the radiation-resistant U87 and U251 cells. Silencing UCA1 or over-expressing miR-873-5p inhibited the survival of U87 and U251 cells, and promoted the cell apoptosis induced by radiation exposure. miR-873-5p was a target gene of UCA1, and UCA1 negatively regulated the expression of miR-873-5p. The inhibition of miR-873-5p could reverse the effect of silencing UCA1 on the radiosensitivity of glioma cells. Silencing UCA1 increased the inhibitory effect of radiation on the glioma cell U251 xenografts. Conclusion Silencing UCA1 inhibits the survival of glioma cells and promotes the cell apoptosis by up-regulating the expression of miR-873-5p, thereby increasing the radiosensitivity of glioma cells.