摘要
目的:探索微小RNA-208b-3p(miR-208b-3p)调节性别决定区Y框蛋白4(SOX4)对非小细胞肺癌细胞增殖、侵袭及上皮间质转化的影响。方法:收集非小细胞肺癌患者癌组织及癌旁组织,并培养人非小细胞肺癌细胞A549、H1299、H460和人正常肺上皮细胞16HBE,qRT-PCR法测定上述组织和细胞miR-208b-3p、SOX4 mRNA表达水平;并将H1299细胞分为对照组、miR-NC组、miR-208b-3p组、miR-208b-3p+pcDNA组、miR-208b-3p+SOX4组;CCK-8法和克隆形成实验测定细胞增殖;Transwell实验测定细胞侵袭;流式细胞仪测定细胞凋亡;Western blot法测定细胞SOX4、Bax、Bcl-2、E-cadherin和Vimentin蛋白表达。结果:与癌旁组织比,肺癌组织中miR-208b-3p表达水平降低,SOX4 mRNA表达水平升高(P<0.05);与16HBE细胞比,A549、H1299、H460细胞中miR-208b-3p表达水平均下降,SOX4 mRNA表达水平均上升(P<0.05);与miR-NC组比,miR-208b-3p组H1299细胞中miR-208b-3p水平、细胞凋亡率、Bax、E-cadherin蛋白表达量升高,而SOX4 mRNA及其蛋白表达水平、OD值、克隆形成率、Bcl-2和Vimentin蛋白表达量均降低,侵袭细胞数减少(P<0.05);与miR-208b-3p+pcDNA组比,miR-208b-3p+SOX4组H1299细胞中细胞凋亡率、Bax、E-cadherin蛋白表达量减少,SOX4 mRNA及其蛋白表达水平、OD值、克隆形成率、Bcl-2及Vimentin蛋白表达量均升高,侵袭细胞数增加(P<0.05)。结论:miR-208b-3p可能通过靶向抑制SOX4表达抑制非小细胞肺癌细胞增殖、侵袭和上皮间质转化。
Objective:To explore the impacts of microRNA-208b-3p(miR-208b-3p)on the proliferation,invasion,and epithelial mesenchymal transition of non-small cell lung cancer cells by regulating sex determining region Y-box 4(SOX4).Methods:Cancer tissues and adjacent tissues were collected from non-small cell lung cancer patients,and human non-small cell lung cancer cells A549,H1299,H460 and human normal lung epithelial cells 16HBE were cultured.The qRT-PCR method was used to measure miR-208b-3p and SOX4 mRNA in the aforementioned tissues and cells.H1299 cells were grouped into control group,miR-NC group,miR-208b-3p group,miR-208b-3p+pcDNA group,and miR-208b-3p+SOX4 group.CCK-8 method and clone formation assay were used to measure cell proliferation.Transwell experiment was used to measure cell invasion.Flow cytometry was used to measure cell apoptosis.In addition,western blot was used to measure the SOX4,Bax,Bcl-2,E-cadherin,and Vimentin proteins in cells.Results:Compared with adjacent tissues,miR-208b-3p was decreased and SOX4 mRNA was increased in lung cancer tissues(P<0.05).Compared with 16HBE cells,miR-208b-3p decreased in A549,H1299,and H460 cells,while SOX4 mRNA increased(P<0.05).Compared with the miR-NC group,the miR-208b-3p group showed increases in miR-208b-3p,apoptosis rate,Bax and E-cadherin proteins in H1299 cells,and decreases in SOX4 mRNA and protein,OD value,clone formation rate,Bcl-2 and Vimentin proteins,and number of invasive cells(P<0.05).Compared with the miR-208b-3p+pcDNA group,the miR-208b-3p+SOX4 group showed decreased apoptosis rate,Bax and E-cadherin proteins in H1299 cells,and an increase in SOX4 mRNA and protein,OD value,clone formation rate,Bcl-2 and Vimentin proteins,and number of invasive cells(P<0.05).Conclusion:MiR-208b-3p may inhibit the proliferation,invasion,and epithelial mesenchymal transition of non-small cell lung cancer cells by targeting the inhibition of SOX4.
作者
丁秋丽
邱琦
彭梅君
刘钧
曾晓
DING Qiuli;QIU Qi;PENG Meijun(Xiaogan Hospital Affiliated to Wuhan University of Science and Technology,Hubei Xiaogan 432000,China)
出处
《河北医学》
2025年第10期1592-1600,共9页
Hebei Medicine
基金
2024年度孝感市自然科学计划立项项目,(编号:XGKJ2024010018)。
