LINC00963靶向miR-1224-5p调控乳腺癌细胞增殖及放射敏感性的分子机制研究

Molecular Mechanism of LINC00963 Targeting miR-1224-5p Regulating Proliferation and Radiosensitivity of Breast Cancer Cells

这项研究探讨长基因间非编码RNA 00963(long intergene noncoding RNA00963,LINC00963)通过靶向miR-1224-5p调控乳腺癌细胞增殖及放射敏感性的分子机制。使用qRT-PCR检测乳腺上皮细胞MCF-10A和乳腺癌细胞(MDA-MB-231、MDA-MB-468、MCF-7)中LINC00963和miR-1224-5p的相对表达情况。将MDA-MB-231细胞分为si-NC组(转染si-NC)、si-LINC00963组(转染si-LINC00963)、miR-NC组(转染miR-NC)、miR-1224-5p组(转染miR-1224-5p)、siLINC00963+anti-miR-NC组(共转染si-LINC00963和anti-miR-NC)、si-LINC00963+anti-miR-1224-5p组(共转染si-LINC00963和anti-miR-1224-5p)。MTT检测细胞增殖情况;Western blot检测细胞周期素D1(CyclinD1)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)蛋白表达情况;克隆实验检测细胞放射敏感性;双荧光素酶报告实验检测LINC00963和miR-1224-5p的靶向关系。在乳腺癌细胞中LINC00963相对表达量明显增加,miR-1224-5p相对表达量显著降低;抑制LINC00963及过表达miR-1224-5p降低乳腺癌细胞增殖活性和细胞存活分数,并下调CyclinD1、PCNA蛋白表达水平。LINC00963靶向调控miR-1224-5p,干扰miR-1224-5p可逆转抑制LINC00963表达对乳腺癌细胞增殖和放射敏感性的影响。LINC00963通过靶向抑制miR-1224-5p增加乳腺癌细胞放射敏感性,并抑制细胞增殖。

This study investigates the molecular mechanisms of LINC00963 targeting miR-1224-5p regulating proliferation and radiosensitivity of breast cancer cells.qRT-PCR was used to detect the relative expression of LINC00963 and miR-1224-5p in breast epithelial cells MCF-10A and breast cancer cells(MDA-MB-231,MDAMB-468,MCF-7).Divide MDA-MB-231 cells into si-NC group(transfected with si-NC),si-LINC00963 group(transfected with si-LINC00963),miR-NC group(transfected with miR-NC),miR-1224-5p group(transfected with miR-1224-5p),si-LINC00963+anti-miR-NC group(co-transfected with si-LINC00963 and anti-miR-NC),siLINC00963+anti-miR-1224-5p group(co-transfected with si-LINC00963 and anti-miR-1224-5p).The expression of LINC00963 and miR-1224-5p was detected by qRT-PCR assay.CyclinD1 and PCNA protein levels were assessed by Western blot assay.Cell proliferation was measured using MTT assay.Cell radiosensitivity was detected by cloning assay.The targeting relationship between LINC00963 and miR-1224-5p was confirmed using a dual luciferase reporter experiment.In breast cancer cells,the relative expression of LINC00963 was significantly increased,and the relative expression of miR-1224-5p was significantly reduced.Inhibition of LINC00963 and overexpression of miR-1224-5p reduced breast cancer cell proliferation activity and cell survival score,and down-regulate CyclinD1,PCNA protein expression.miR-1224-5p acted as a target of LINC00963.Interference miR-1224-5p reversed the effects of LINC00963 knockdown on breast cancer cell proliferation and radiosensitivity.LINC00963 might improve breast cancer cell radiosensitivity and suppress cell proliferation by targeting miR-1224-5p.