LINC00667靶向miR-154-5p调控乳腺癌细胞增殖及凋亡的分子机制研究  

Molecular mechanism of LINC00667 regulating breast cancer cell proliferation and apoptosis by targeting miR-154-5p

作　　者：杨瑞玲[1] 桑梅香[2] 耿翠芝[3] 付俊勇 YANG Ruiling;SANG Meixiang;GENG Cuizhi;FU Junyong(Breast Surgery,Handan Central Hospital,Handan,Hebei 056001,China;The Fourth Hospital of Hebei Medical University,Scientific Research Center,Shijiazhuang,Hebei 050011,China;Breast Center,the Fourth Hospital of Hebei Medical University,Shijiazhuang,Hebei 050011,China;Emergency Department,North Hospital,Jizhong Energy Fengfeng Mining Bureau General Hospital,Handan,Hebei 056299,China)

机构地区：[1]邯郸市中心医院乳腺外科,河北邯郸056001 [2]河北医科大学第四医院科研中心,河北石家庄050011 [3]河北医科大学第四医院乳腺中心,河北石家庄050011 [4]冀中能源峰峰矿务局总医院北院区急诊科,河北邯郸056299

出　　处：《中国优生与遗传杂志》2022年第7期1120-1125,共6页Chinese Journal of Birth Health & Heredity

摘　　要：目的 探讨LINC00667调控乳腺癌细胞增殖及凋亡的可能作用机制。方法 收集2020年5月至2020年8月邯郸市中心医院收治的51例乳腺癌患者的癌组织及其癌旁组织标本,采用qRT-PCR法检测LINC00667、miR-154-5p的表达量;体外培养人乳腺癌细胞T-47D,si-NC、si-LINC00667、miR-NC、miR-154-5p mimics分别转染至T-47D细胞,si-LINC00667和anti-miR-NC、si-LINC00667和anti-miR-154-5p分别共转染至T-47D细胞;MTT法、平板克隆形成实验与流式细胞术分别检测细胞增殖、克隆形成及凋亡;双荧光素酶报告实验检测miR-154-5p过表达对野生型载体WT-LINC00667与突变型载体MUT-LINC00667荧光素酶活性的影响;Western blot检测Bax、Bcl-2蛋白表达量。结果与癌旁组织比较,乳腺癌组织中LINC00667的表达量升高(P<0.05),miR-154-5p的表达量降低(P<0.05);转染si-LINC00667或转染miR-154-5p mimics后细胞增殖抑制率、细胞凋亡率和Bax蛋白水平升高(P<0.05),细胞克隆形成数减少(P<0.05),Bcl-2蛋白水平降低(P<0.05);miR-154-5p过表达可抑制WT-LINC00667的荧光素酶活性(P<0.05);共转染si-LINC00667和anti-miR-154-5p可减弱转染si-LINC00667对细胞生物学行为的作用。结论 抑制LINC00667表达可通过促进miR-154-5p表达而减弱乳腺癌细胞增殖及克隆形成能力,并可诱导细胞凋亡。Objective To explore the possible mechanism of LINC00667 regulating breast cancer cell proliferation and apoptosis. Methods The cancer tissue and adjacent tissue samples of 51 breast cancer patients admitted to our hospital from May 2020 to August 2020 were collected, and the expression levels of LINC00667 and miR-154-5p were detected by qRT-PCR method. Human breast cancer cells T-47D were cultured in vitro, si-NC, si-LINC00667, miR-NC, and miR-154-5p mimics were transfected into T-47D cells, respectively. si-LINC00667 and anti-miR-NC, si-LINC00667 and anti-miR-154-5p were co-transfected into T-47D cells, respectively. Cell proliferation, clone formation and apoptosis were detected by MTT method, plate colony formation assay and flow cytometry, respectively. The effect of miR-154-5p overexpression on the luciferase activity of wild-type vector WT-LINC00667 and mutant vector MUT-LINC00667 was detected by dual-luciferase reporter assay. Western blot was used to detect the expression of Bax, Bcl-2 protein. Results Compared with the adjacent tissue, the expression of LINC00667 in breast cancer tissue was increased(P<0.05), while the expression of miR-154-5p was decreased(P<0.05). After transfection of si-LINC00667 or miR-154-5p mimics, the inhibition rate of cell proliferation, the rate of apoptosis and the protein level of Bax were increased(P<0.05), while the number of cell clones was decreased(P<0.05),the protein level of Bcl-2 was decreased(P<0.05). Overexpression of miR-154-5p could inhibit the luciferase activity of WT-LINC00667(P<0.05). Co-transfection of si-LINC00667 and anti-miR-154-5p attenuated the effect of transfection of si-LINC00667 on the biological behavior of cells. Conclusion Inhibition of LINC00667 expression could attenuate breast cancer cell proliferation and clone formation by promoting miR-154-5p expression, and could induce apoptosis.

关 键 词：乳腺癌 LINC00667 miR-154-5p 细胞增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]