摘要
目的 探究miR-326通过调控EphB3的表达,抑制乳腺癌侵袭转移的分子机制。方法 RTFQ-PCR用于检测miR-326在正常乳腺上皮细胞及乳腺癌细胞中的表达情况及miR-326过表达质粒的转染效率。EdU细胞增殖实验、Transwell实验用于检测不同分组细胞增殖、迁移和侵袭能力的变化。双荧光素酶实验用于验证miR-326与EphB3是否存在结合位点。Western blot法检测过表达miR-326后乳腺癌细胞中EphB3的表达情况。结果 RTFQ-PCR结果显示,miR-326在乳腺癌细胞中低表达,且转染成功(P<0.05)。EdU增殖实验和Transwell实验结果表明,过表达miR-326能够抑制乳腺癌细胞的增殖、迁移和侵袭能力(P<0.05)。双荧光素酶实验结果显示,miR-326可以与EphB3的3′-UTR相互结合(P<0.05)。Western blot和Transwell实验结果显示,miR-326可以负向调控EphB3抑制乳腺癌细胞的侵袭转移(P<0.05)。结论 miR-326在乳腺癌的发生发展过程中起着抑癌基因的作用,并通过调控EphB3的表达抑制乳腺癌细胞的侵袭和转移。
Aim To investigate the molecular mechanism of miR-326 inhibiting breast cancer invasion and metastasis by regulating EphB3 expression.Methods RTFQ-PCR was used to examine the expression of miR-326 in normal breast epithelial cells and breast cancer cells and the transfection efficiency of miR-326 overexpression plasmid.EdU cell proliferation assay and Transwell assay were used to examine the changes in proliferation,migration and invasion ability of different subgroups of cells.Dual luciferase assay was used to verify the presence of binding sites for miR-326 and EphB3.Western blot was used to detect the expression of EphB3 in breast cancer cells after overexpression of miR-326.Results RTFQ-PCR results showed that miR-326 was lowly expressed in breast cancer cells and successfully transfected(P<0.05).EdU proliferation assay and Transwell assay results showed that overexpression of miR-326 in breast cancer cells inhibited proliferation,migration and invasive ability(P<0.05).The results of dual luciferase assay showed that miR-326 could interact with the 3′-UTR of EphB3(P<0.05).Western blot and Transwell assays showed that miR-326 could negatively regulate EphB3 to inhibit invasive metastasis of breast cancer cells(P<0.05).Conclusions MiR-326 acts as a cancer suppressor genes in the development of breast cancer and suppresses the invasion and metastasis of breast cancer cells by regulating the expression of EphB3.
作者
陈新璐
陈琳
左伟
赵燕乔
刘璐
李洪利
李承德
尹崇高
CHEN Xin-lu;CHEN Lin;ZUO Wei;ZHAO Yan-qiao;LIU Lu;LI Hong-li;LI Cheng-de;YIN Chong-gao(Dept of Pathology,Weifang Medical University,Weifang Shandong 261053,China;Affiliated Hospital of Weifang Medical College,Weifang Medical University,Weifang Shandong 261053,China;Medical Research Center,Weifang Medical University,Weifang Shandong 261053,China;Dept of Clinical Pharmacy,Weifang Medical University,Weifang Shandong 261053,China;Surgical Nursing College of Nursing,Weifang Medical University,Weifang Shandong 261053,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2023年第4期665-672,共8页
Chinese Pharmacological Bulletin
基金
山东省自然科学基金项目(No ZR2019MH033)
山东省高等学校青创人才引育计划(No 205)。
