摘要
目的 探讨miR-34a-3p对胰腺癌细胞的作用及可能机制。方法 RT-qPCR方法检测miR-34a-3p在人胰腺导管腺癌细胞SW1990和胰腺导管上皮细胞HPDE中的表达情况;转染miR-34a-3p mimics后,采用CCK-8法检测细胞增殖;Transwell小室实验检测细胞侵袭情况;Western blot检测细胞上皮-间质转化情况;进一步用生物信息学预测miR-34a-3p的下游靶基因,并用荧光素酶报告基因实验验证;然后采用Western blot检测靶蛋白以及靶蛋白下游调控蛋白的表达。结果 RT-qPCR检测发现miR-34a-3p在SW1990中的表达低于HPDE细胞。miR-34a-3p在SW1990细胞中过表达后,SW1990细胞增殖受到抑制、侵袭能力降低、E-cadherin表达上调、N-cadherin表达下调。生物信息学网站预测、双荧光素酶报告基因实验和Western blot证实Smad2是miR-34a-3p的靶蛋白,同时发现其下游蛋白TGF-β表达也显著下调。结论 miR-34-3p在胰腺癌细胞中的表达下调,miR-34-3p能够抑制胰腺癌细胞增殖、侵袭转移和上皮-间质转化,该作用与靶向调控Smad2/TGF-β信号通路有关。
Objective To investigate the effect of miR-34A-3p on pancreatic cancer cells and its possible mechanism. Methods RT-qPCR was used to detect the expression of miR-34A-3p in human pancreatic ductal adenocarcinoma cell SW1990 and pancreatic ductal epithelial cell HPDE. After transfection with miR-34A-3p mimics, cell proliferation was detected by CCK-8 assay. Cell invasion was detected by Transwell assay and epithelial-mesenchymal transformation was detected by Western blot. The downstream target genes of miR-34A-3p were further predicted by bioinformatics and verified by luciferase reporter gene assay. Western blot was used to detect the expression of target protein and downstream regulatory protein. Results RT-qPCR results showed that the expression of miR-34A-3p in SW1990 was lower than that in HPDE cells. After overexpression of miR-34A-3p in SW1990 cells, the proliferation of SW1990 cells was inhibited, while the cell invasion ability was reduced, the expression of E-cadherin was up-regulated, and the expression of N-cadherin was down-regulated. Smad2 was confirmed to be the target protein of miR-34a-3p. At the same time, it was found that the expression of its downstream protein TGF-β was also significantly down-regulated. Conclusion The expression of miR-34a-3p is down-regulated in pancreatic cancer cells. miR-34a-3p can inhibit the proliferation, invasion,metastasis and epithelial-mesenchymal transition of pancreatic cancer cells, which is related to the targeted regulation of Smad2/TGF-β signaling pathway.
作者
纪大伟
赵明程
陈广泉
李隆钢
蔡景钰
JI Da-wei;ZHAO Ming-cheng;CHEN Guang-quan;LI Long-gang;CAI Jing-yu(Department of General Surgery,Shenyang Fourth People's Hospital,Shenyang 110030,China)
出处
《解剖科学进展》
CAS
2022年第2期149-152,共4页
Progress of Anatomical Sciences
基金
辽宁省自然科学基金(20180551041)。
