摘要
长链非编码RNA LINC00342已被证实在多种癌症中参与重要生物学功能。然而,LINC00342在乳腺癌中的作用和机制尚不清楚。在该研究中,选取28例乳腺癌患者肿瘤组织和癌旁正常组织,乳腺癌细胞和正常乳腺上皮细胞,用qRT-PCR检测LINC00342、miR-505-3p和磷酸甘油酸激酶1(PGK1)mRNA的表达情况。Western blot检测PGK1蛋白的表达情况。将乳腺癌细胞MCF-7分为NC组(空白)、si-LINC00342组(转染si-LINC00342)、si-NC组(转染si-NC)、miR-505-3p组(转染miR-505-3p模拟物)、miR-NC组(转染miR-NC)、si-PGK1组(转染si-PGK1)、si-NC组(转染si-NC)、si-LINC00342+pcDNA-PGK1组(同时转染si-LINC00342与pcDNA-PGK1)和siLINC00342+pcDNA组(同时转染si-LINC00342与pcDNA)。利用Western blot检测PGK1、迁移侵袭标志物基质金属蛋白酶MMP2和MMP9的表达情况,MTT法检测细胞增殖能力,Transwell法检测细胞迁移和侵袭数量。双荧光素酶活性实验检测miR-505-3p与LINC00342、PGK1之间的靶向结合。结果显示,LINC00342、PGK1 mRNA和PGK1蛋白在乳腺癌组织和细胞中显著上调,miR-505-3p显著下调。干扰LINC00342、过表达miR-505-3p或干扰PGK1均能抑制乳腺癌细胞增殖、迁移和侵袭,以及MMP2和MMP9蛋白表达。此外,LINC00342可直接靶向miR-505-3p调控PGK1表达,高表达PGK1可逆转LINC00342低表达对MCF-7增殖、迁移和侵袭的抑制效果。该研究提示,干扰LINC00342表达可能通过靶向miR-505-3p调控PGK1的表达,抑制乳腺癌细胞的增殖、迁移和侵袭。
Long non-coding RNA LINC00342 has been shown to be involved in important biological functions in a variety of cancers.However,the role and mechanism of LINC00342 in breast cancer remain unclear.This study selected 28 paired tumor tissue and adjacent normal tissue from breast cancer patients,as well as breast cancer cells and normal breast epithelial cells.The expressions of LINC00342,miR-505-3p,PGK1(phosphoglycerate kinase 1)mRNA were detected by qRT-PCR.The protein expression of PGK1 was detecetd by Western blot.The breast cancer MCF-7 cells were divided into NC group(blank),si-LINC00342 group(transfected with siLINC00342),si-NC group(transfected with si-NC),miR-505-3p group(transfected with miR-505-3p mimic),miRNC group(transfected with miR-NC),si-PGK1 group(transfected with si-PGK1),si-NC group(transfected with si-NC),si-LINC00342+pcDNA-PGK1 group(simultaneous transfection with si-LINC00342 and pcDNA-PGK1),si-LINC00342+pcDNA group(simultaneous transfection with si-LINC00342 and pcDNA).Western blot was used to detect the expression of PGK1,MMP2(matrix metalloprotease 2),and MMP9 protein.The MTT method was implemented to monitor the cell proliferation ability,and the Transwell method was employed to assess the number of cell migration and invasion.The dual luciferase activity assay detects the targeted binding between miR-505-3p,LINC00342 and PGK1.The results showed that LINC00342,PGK1 mRNA,and PGK1 protein were significantly up-regulated in breast cancer tissues and cells,while miR-505-3p was significantly down-regulated.LINC00342interference,miR-505-3p overexpression or PGK1 knockdown could inhibit the proliferation,migration,invasion,and the expression of MMP2 and MMP9 proteins in breast cancer cells.In addition,LINC00342 can directly target miR-505-3p to regulate the expression of PGK1,and high expression of PGK1 can reverse the inhibitory effect of low expression of LINC00342 on the proliferation,migration,and invasion of MCF-7.This study suggests that interference with the expression of LINC00342 may regulate the expression of PGK1 by targeting miR-505-3p to inhibit the proliferation,migration,and invasion of breast cancer cells.
作者
郭莹叶
GUO Yingye(School of Basic Medicine,Jiangxi Medical College,Shangrao 334000,China)
出处
《中国细胞生物学学报》
CAS
CSCD
2023年第2期193-202,共10页
Chinese Journal of Cell Biology
