沉默CD24对乳腺癌MCF‑7细胞增殖和侵袭的影响及机制的初步研究

Effect and mechanism of silencing CD24 gene on proliferation and invasion of breast cancer cells MCF-7:a primary study

目的探讨CD24在乳腺癌中的表达及靶向下调CD24对乳腺癌MCF-7细胞增殖和侵袭的影响及机制。方法通过生物信息学网站分析TCGA数据库中乳腺癌数据集CD24 mRNA和蛋白的表达及与乳腺癌患者生存期的关系。采用qRT-PCR和Western blot方法检测人正常乳腺细胞系MCF-10A及乳腺癌细胞系MCF-7、MDA-MB-231和T-47D中CD24的表达;利用siRNA干扰MCF-7细胞中CD24的表达,设置空白组(未进行转染)、si-NC组(转染无义序列)、siCD24组(转染CD24干扰物);CCK-8法检测各组乳腺癌细胞的增殖能力;Transwell实验检测各组乳腺癌细胞的侵袭能力;Western blot检测各组乳腺癌细胞中TFPI-2的表达。结果CD24在乳腺癌中的表达升高,表达水平与患者的生存期显著相关(P<0.05);与人乳腺正常细胞系MCF-10A相比,CD24在MCF-7、MDA-MB-231和T-47D乳腺癌细胞系中表达升高(P<0.05);CCK-8结果显示,干扰CD24的表达可以抑制乳腺癌细胞MCF-7的增殖能力。Transwell结果显示,干扰CD24的表达可以抑制乳腺癌细胞MCF-7的侵袭能力(P<0.05)。Western blot结果显示,CD24表达下调后,siCD24组的乳腺癌细胞中TFPI-2蛋白表达升高(P<0.05)。结论CD24在乳腺癌中高表达,表达水平与患者的生存期显著相关,抑制CD24表达可以抑制乳腺癌细胞的增殖能力和侵袭能力,其机制可能与负向调控TFPI-2基因的表达相关。

Objective To investigate the expression of CD24 in breast cancer and the effect and mechanism of down-regulating CD24 on the proliferation and invasion of breast cancer cells MCF-7.Methods The expression of CD24 mRNA and protein in breast cancer dataset in TCGA database and its relationship with the survival time of breast cancer patients were analyzed by bioinformatics website.The expression of CD24 in normal human breast cell line MCF-10A and breast cancer cell lines MCF-7,MDA-MB-231 and T-47D was detected by qRT-PCR and Western blot.siRNA was used to interfere with the expression of CD24 in MCF-7 cells.The blank group(without transfection),si-NC(transfected senseless sequence),and siCD24(transfected CD24 interference)were set in the experiment.CCK-8 and Transwell experiments were used to detect the proliferation and invasion ability of cells.Western blot was used to detect the expression of TFPI-2 in breast cancer cells.Results The expression of CD24 in breast cancer was increased,and the expression level was significantly correlated with the survival time of patients(P<0.05).Compared with normal human breast cell line MCF-10A,the expression of CD24 in MCF-7,MDA-MB-231 and T-47D breast cancer cells was increased(P<0.05).CCK-8 assay and Transwell results showed that CD24 interference could inhibit the proliferation and invasion of breast cancer cell MCF-7(P<0.05).Western blot results showed that the expression of TFPI-2 protein in breast cancer cells increased in siCD24 group after the down-regulation of CD24 expression(P<0.05).Conclusion CD24 is highly expressed in breast cancer,and the expression level is significantly correlated with the survival time of patients.Inhibition of CD24 expression can inhibit the proliferation and invasion of breast cancer cells,and the mechanism may be related to the negative regulation of TFPI-2 gene expression.