miR-431-5p通过调控AKT1抑制胰腺癌细胞增殖、迁移、侵袭和促进凋亡研究

汪俊州; 王义刚; 孙雅轩

doi:10.13898/j.cnki.issn.1000-2200.2022.06.004

miR-431-5p通过调控AKT1抑制胰腺癌细胞增殖、迁移、侵袭和促进凋亡研究

Study on the miR-431-5p inhibiting the proliferation, migration, invasion and promoting the apoptosis of pancreatic cancer cells by regulating AKT1

摘要

摘要:
目的研究miR-431-5p对胰腺癌细胞增殖、迁移、侵袭和凋亡的影响和潜在的分子机制。
方法qRT-PCR检测人正常胰腺上皮细胞hTERT-HPNE和胰腺癌细胞CFPAC-1和PANC-1中miR-431-5p表达水平。转染miR-431-5p模拟物至胰腺癌CFPAC-1细胞中过表达miR-431-5p后，MTT法测定细胞增殖活性，Transwell检测细胞迁移和侵袭，流式细胞术检测细胞凋亡，Western blotting检测细胞中Cyclin D1、p21、Bax、Bcl-2、E-cadherin、MMP-2和丝氨酸/苏氨酸蛋白激酶1（AKT1）蛋白表达。双荧光素酶报告基因实验验证miR-431-5p与AKT1的调控关系。
结果与hTERT-HPNE细胞相比，胰腺癌细胞PANC-1和CFPAC-1中miR-431-5p表达量降低（P < 0.05）。过表达miR-431-5p后，CFPAC-1细胞活性、迁移和侵袭细胞数降低，凋亡率升高，Cyclin D1、Bcl-2和MMP-2蛋白表达水平降低，p21、Bax和E-cadherin蛋白表达水平升高，差异均有统计学意义（P < 0.05~P < 0.01）。双荧光素酶报告基因实验结果显示，miR-431-5p在CFPAC-1细胞中靶向负调控AKT1表达。同时过表达miR-431-5p和AKT1后，CFPAC-1细胞活性、迁移和侵袭细胞数升高，凋亡率降低，Cyclin D1、Bcl-2和MMP-2蛋白表达水平升高，p21、Bax和E-cadherin蛋白表达水平降低，差异均有统计学意义（P < 0.05）。
结论miR-431-5p通过靶向抑制AKT1降低胰腺癌CFPAC-1细胞的增殖、迁移和侵袭能力，并促进细胞凋亡，可能是胰腺癌的潜在分子治疗靶点。

Abstract:
ObjectiveTo investigate the effects of miR-431-5p on the proliferation, migration, invasion and apoptosis of pancreatic cancer cells, and its underlying mechanism.
MethodsThe qRT-PCR was used to detect the expression levels of miR-431-5p in human normal pancreatic epithelial cells hTERT-HPNE and pancreatic cancer cells CFPAC-1 and PANC-1. After the miR-431-5p mimics were transfected into the pancreatic cancer CFPAC-1 cells to overexpress miR-431-5p, the MTT assay was used to detect the cell proliferation activity, the Transwell assay was used to detect the cell migration and invasion, the flow cytometry was used to detect the apoptosis, and the Western blotting was used to detect the expression levels of Cyclin D1, p21, Bax, Bcl-2, E-cadherin, MMP-2 and AKT1 proteins. The regulatory relationship between miR-431-5p and AKT1 was investigated using the dual luciferase reporter gene experiment.
ResultsCompared with hTERT-HPNE cells, the expression levels of miR-431-5p in pancreatic cancer cell PANC-1 and CFPAC-1 significantly decreased(P < 0.05). After the miR-431-5p in CFPAC-1 cells was overexpressed, the cells activity and number of migrating and invading cells decreased(P < 0.05), the apoptosis rate increased(P < 0.05), the expression levels of Cyclin D1, Bcl-2 and MMP-2 protein decreased(P < 0.05), and the expression levels of p21, Bax and E-cadherin protein increased(P < 0.05 to P < 0.01). The results of the dual luciferase reporter gene experiment showed that the miR-431-5p negatively regulated the AKT1 expression in CFPAC-1 cells. After the miR-431-5p and AKT1 in CFPAC-1 cells were overexpressed, the CFPAC-1 cell activity, the number of migrating and invading cells increased, the apoptosis rate decreased, the protein expression levels of Cyclin D1, Bcl-2 and MMP-2 increased, while the protein expression levels of p21, Bax and E-cadherin decreased, and the differences of which were statistically significant(P < 0.05).
ConclusionsThe miR-431-5p can inhibit the proliferation, migration and invasion of pancreatic cancer CFPAC-1 cells, and promote apoptosis by targeting AKT1. It may be a potential molecular therapeutic target for pancreatic cancer.

HTML全文

参考文献(17)

施引文献

资源附件(0)