摘要
目的探讨miR-26b-5p对非小细胞肺癌细胞增殖、迁移和侵袭的影响及其作用机制。方法培养人正常肺上皮细胞BEAS-2B和非小细胞肺癌细胞A549,通过qPCR检测BEAS-2B和A549细胞中miR-26b-5p和VEGF mRNA的表达水平。通过Western Blot检测BEAS-2B和A549细胞中E-cadherin、Vimentin和VEGF蛋白的表达水平。生物信息学预测VEGF与miR-26b-5p的靶向关系。使用脂质体法将NC-mimic和miR-26b-5p-mimic转染A549细胞,通过CCK8法、EDU实验检测miR-26b-5p对细胞生长的影响;通过划痕实验和Transwell侵袭实验检测miR-26b-5p对细胞迁移和侵袭的影响;通过Western Blot检测对E-cadherin、Vimentin和VEGF蛋白表达的影响。结果与BEAS-2B细胞相比,在A549细胞中,miR-26b-5p表达水平显著下降(P<0.05),VEGF的mRNA和蛋白表达水平显著上升(P<0.05),E-cadherin蛋白的表达水平显著下降(P<0.05),而Vimentin的蛋白表达水平显著上升(P<0.05)。与NC-mimic组相比,miR-26b-5p-mimic组在24h、48h以及72h时细胞吸光值显著减少(P<0.05),24h划痕距离显著增加(P<0.05),24h侵袭细胞数显著减少(P<0.05),E-cadherin蛋白表达显著上调(P<0.05)、Vimentin和VEGF蛋白表达显著下调(均P<0.05)。结论过表达miR-26b-5p可以通过靶向抑制VEGF的表达,从而抑制非小细胞肺癌A549细胞的增殖、迁移和侵袭。
Objective To investigate the effect of miR-26b-5p on the growth,migration,and invasion of non-small cell lung cancer cells and its mechanism.Methods The expression levels of miR-26b-5p and VEGF mRNA in BEAS-2B and A549 cells were detected by qPCR.The expression levels of E-cadherin,Vimentin,and VEGF proteins in BEAS-2B and A549 cells were detected by Western blot.Bioinformatics predicted the targeting relationship between VEGF and miR-26b-5p.NC mimics and miR-26b-5p-mimics were transfected into A549 cells by liposome method,and the effects on cell growth were detected by CCK8 method and EDU experiment;The effects on cell migration and invasion were detected by scratch assay and Transwell invasion assay;The expression of E-cadherin,Vimentin,and VEGF was detected by Western blot.Results Compared with BEAS-2B cells,in A549 cells,the expression level of miR-26b-5p was significantly decreased(P<0.05),the mRNA and protein expression levels of VEGF were significantly increased(P<0.05),the expression level of E-cadherin protein was significantly decreased(P<0.05),and the protein expression level of Vimentin was significantly increased(P<0.05).Compared with NC mimic group,miR-26b-5p-mimic group significantly decreased the absorbance value of cells at 24h,48h and 72h(P<0.05),significantly increased the scratch distance at 24h(P<0.05),significantly reduced the number of invasive cells at 24h(P<0.05),significantly upregulated the expression of E-cadherin protein(P<0.05),and significantly downregulated the expression of vimentin and VEGF protein(all P<0.05).Conclusion Overexpression of miR-26b-5p can inhibit the growth,migration,and invasion of non-small cell lung cancer A549 cells by targeting the expression of VEGF.
作者
柳鑫
姜晓丽
马雪梅
LIU Xin;JIANG Xiaoli;MA Xuemei(Department of Respiratory Medicine,The First Affiliated Hospital of Jiamusi University,Jiamusi,Heilongjiang 154000,China)
出处
《临床肺科杂志》
2024年第3期375-381,共7页
Journal of Clinical Pulmonary Medicine
基金
佳木斯大学附属第一医院国家重点专科建设项目(No.GJ202302)。
